Single-cell RNA sequencing reveals the pro-inflammatory roles of liver-resident Th1-like cells in primary biliary cholangitis.

Primary biliary cholangitis (PBC) is a chronic autoimmune liver disease characterized by multilineage immune dysregulation, which subsequently causes inflammation, fibrosis, and even cirrhosis of liver. Due to the limitation of traditional assays, the local hepatic immunopathogenesis of PBC has not been fully characterized. Here, we utilize single-cell RNA sequencing technology to depict the immune cell landscape and decipher the molecular mechanisms of PBC patients. We reveal that cholangiocytes and hepatic stellate cells are involved in liver inflammation and fibrosis. Moreover, Kupffer cells show increased levels of inflammatory factors and decreased scavenger function related genes, while T cells exhibit enhanced levels of inflammatory factors and reduced cytotoxicity related genes. Interestingly, we identify a liver-resident Th1-like population with JAK-STAT activation in the livers of both PBC patients and murine PBC model. Finally, blocking the JAK-STAT pathway alleviates the liver inflammation and eliminates the liver-resident Th1-like cells in the murine PBC model. In conclusion, our comprehensive single-cell transcriptome profiling expands the understanding of pathological mechanisms of PBC and provides potential targets for the treatment of PBC in patients.

Electrochemiluminescence immunoassay system based on PCN-224-Mn and gold-platinum bimetallic nanoflowers for sensitive detection of ochratoxin A.

In this work, a novel Electrochemiluminescence Immunosensor was constructed using PCN-224-Mn and gold-platinum nanoflowers (AuPt NFs) for the ultrasensitive detection of ochratoxin A (OTA). PCN-224 modified with Mn (II) was synthesized as a probe material. The interaction efficiency of PCN-224 with S2O82- was also greatly improved. AuPt NFs were used as the substrate material for the electrodes. It has favorable biocompatibility, large specific surface area and can bind more antigen. Also greatly increased the electroactive surface area and conductivity of the electrode. OTA was detected using a competitive immunoassay strategy, in which OTA in the sample competes with the encapsulated antigen for a finite number of antibodies. ECLIA for the detection of OTA was designed to be highly sensitive, with a linear range from 0.0002 ng mL-1 to 1000 ng mL-1 and a LOD as low as 0.067 pg mL-1. In addition, it was evident from the electrochemical analyses that PCN-224-Mn had a stronger and more stable ECL signal compared to the plain PCN-224. The successful preparation of specific, sensitive and reproducible ECL immunosensors confirms the great promise for the detection of OTA or other small molecule mycotoxins.

Antenna effect enhanced ECL immunoassay using microfloral europium porphyrin coordination polymers based on Eu3+ and TCPP for the detection of chloramphenicol in foods.

The "antenna effect" is one of the most important energy transfer modes in lanthanide light-emitting polymers. In this study, novel luminescent nanostructured coordination polymers (Eu-PCP) were synthesized in one step using Eu3+ as the central metal ion and 5,10,15,20-tetrakis (4-carboxyphenyl) porphyrin (TCPP) as the organic ligand. The unique "antenna effect" observed between Eu3+ and TCPP leads to a substantial improvement in the electrochemiluminescence (ECL) emission efficiency. Eu-PCP exhibits good cathodic ECL characteristics. Additionally, Au@SnS2 nanosheets exhibit favorable electrical conductivity, biocompatibility, and a significant specific surface area. This makes them a suitable choice as substrate materials for the modification of electrode surfaces and capturing antigens. Being well known, the development of sensitive and rapid methods to detect chloramphenicol is essential for food safety. Based on this, we report a novel competitive electrochemiluminescence immunoassay to achieve ultra-sensitive and highly specific detection of chloramphenicol. The linear range was 0.0002-500 ng mL-1 and the detection limit was 0.09 pg mL-1. Apart from that, the experimental results proved that it provided a new analytical tool for the detection of antibiotic residues in food safety.

Adhesion GPCR ADGRE2 Maintains Proteostasis to Promote Progression in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is an aggressive and heterogeneous hematologic malignancy. In elderly patients, AML incidence is high and has a poor prognosis due to a lack of effective therapies. G protein-coupled receptors (GPCR) play integral roles in physiologic processes and human diseases. Particularly, one third of adhesion GPCRs, the second largest group of GPCRs, are highly expressed in hematopoietic stem and progenitor cells or lineage cells. Here, we investigate the role of adhesion GPCRs in AML and whether they could be harnessed as antileukemia targets. Systematic screening of the impact of adhesion GPCRs on AML functionality by bioinformatic and functional analyses revealed high expression of ADGRE2 in AML, particularly in leukemic stem cells, which is associated with poor patient outcomes. Silencing ADGRE2 not only exerts antileukemic effects in AML cell lines and cells derived from patients with AML in vitro, but also delays AML progression in xenograft models in vivo. Mechanistically, ADGRE2 activates phospholipase Cß/protein kinase C/MEK/ERK signaling to enhance the expression of AP1 and transcriptionally drive the expression of DUSP1, a protein phosphatase. DUSP1 dephosphorylates Ser16 in the J-domain of the co-chaperone DNAJB1, which facilitates the DNAJB1-HSP70 interaction and maintenance of proteostasis in AML. Finally, combined inhibition of MEK, AP1, and DUSP1 exhibits robust therapeutic efficacy in AML xenograft mouse models. Collectively, this study deciphers the roles and mechanisms of ADGRE2 in AML and provides a promising therapeutic strategy for treating AML. Significance: Increased expression of the adhesion GPCR member ADGRE2 in AML supports leukemia stem cell self-renewal and leukemogenesis by modulating proteostasis via an MEK/AP1/DUSP1 axis, which can be targeted to suppress AML progression.

Ultrasensitive detection of zearalenone based on electrochemiluminescent immunoassay with Zr-MOF nanoplates and Au@MoS2 nanoflowers.

In this work, an effective competitive-type electrochemiluminescence (ECL) immunosensor was constructed for zearalenone determination by using Zr-MOF nanoplates as the ECL luminophore and Au@MoS2 nanoflowers as the substrate material. Zr-MOF have an ultra-thin sheet-like structure that accelerates the transfer of electrons, ions and co-reactant intermediates, which exhibited strong and stable anodic luminescence. The three-dimensional Au@MoS2 nanoflowers would form a thin film modification layer on the glassy carbon electrode (GCE). And its good electrical conductivity and higher specific surface area utilization further improving the sensitivity of the ECL immunosensor. Under the optimized conditions, the proposed immunosensor exhibited satisfactory stability, sensitivity and accuracy, and its ECL signal was proportional to the logarithm of ZEN concentration (0.0001-100 ng/mL) and the limit of detection (LOD) was 0.034 pg/mL. In addition, the results of recovery experiment acquired for wheat flour and pig urine samples further proved the feasibility of the immunosensor for the detection of real samples, indicating its potential for ultrasensitive detection of ZEN.

Single-cell RNA sequencing reveals the immunoregulatory roles of PegIFN-α in patients with chronic hepatitis B.

BACKGROUND AND AIMS: Chronic hepatitis B (CHB) is caused by HBV infection and affects the lives of millions of people worldwide by causing liver inflammation, cirrhosis, and liver cancer. Interferon-alpha (IFN-α) therapy is a conventional immunotherapy that has been widely used in CHB treatment and achieved promising therapeutic outcomes by activating viral sensors and interferon-stimulated genes (ISGs) suppressed by HBV. However, the longitudinal landscape of immune cells of CHB patients and the effect of IFN-α on the immune system are not fully understood. APPROACH AND RESULTS: Here, we applied single-cell RNA sequencing (scRNA-seq) to delineate the transcriptomic landscape of peripheral immune cells in CHB patients before and after PegIFN-α therapy. Notably, we identified three CHB-specific cell subsets, pro-inflammatory (Pro-infla) CD14+ monocytes, Pro-infla CD16+ monocytes and IFNG+ CX3CR1- NK cells, which highly expressed proinflammatory genes and positively correlated with HBsAg. Furthermore, PegIFN-α treatment attenuated percentages of hyperactivated monocytes, increased ratios of long-lived naive/memory T cells and enhanced effector T cell cytotoxicity. Finally, PegIFN-α treatment switched the transcriptional profiles of entire immune cells from TNF-driven to IFN-α-driven pattern and enhanced innate antiviral response, including virus sensing and antigen presentation. CONCLUSIONS: Collectively, our study expands the understanding of the pathological characteristics of CHB and the immunoregulatory roles of PegIFN-α, which provides a new powerful reference for the clinical diagnosis and treatment of CHB.

Co-exposure of polystyrene microplastics influence cadmium trophic transfer along the "lettuce-snail" food chain: Focus on leaf age and the chemical fractionations of Cd in lettuce.

Cadmium (Cd) and polystyrene microplastics (PS) co-contamination always occurs in environment; however, the trophic transfer of Cd and PS is still poorly understood. A hydroponic experiment was conducted to investigate the behavior of Cd in lettuce, together with the root or foliar exposure of different sized PS. Accumulation and chemical form distributions of Cd in leaves were distinguished into young and mature leaves. Subsequently, a 14-day snail feeding experiment was performed. Data showed that Cd accumulation in roots, rather than in leaves, are significantly affected by PS coexistence. However, mature leaves had a higher Cd content than young leaves under the root exposure of PS, while a reverse effect was observed in the foliar exposure. There existed a positive correlation between the food-chain transfer associated Cd (CdFi+Fii+Fiii) in mature leaves and Cd content in snail soft tissue (r = 0.705, p < 0.001), but not in young leaves. Though no bio-amplification of Cd in food chain was observed, an increase of Cd transfer factor (TF) from lettuce to snail was noted in the root exposure of 5 µm PS and the foliar exposure of 0.2 µm PS. Moreover, we observed a highest increase rate of 36.8 % in TF values from lettuce to snail viscera, and a chronic inflammatory response in snail stomach tissue. Therefore, more attentions should be paid to study the ecological risks of heavy metals and microplastics co-contamination in environment.

Dlk1 maintains adult mice long-term HSCs by activating Notch signaling to restrict mitochondrial metabolism.

BACKGROUND: Adult hematopoietic stem cells (HSCs) homeostasis is critically important in maintaining lifelong hematopoiesis. However, how adult HSCs orchestrate its homeostasis remains not fully understood. Imprinted gene Dlk1 has been shown to play critical role in mouse embryonic hematopoiesis and in regulation of stem cells, but its physiological roles in adult HSCs are unknown. METHODS: We performed gene expression analysis of Dlk1, and constructed conditional Dlk1 knockout (KO) mice by crossing Mx1 cre mice with Dlkflox/flox mice. Western blot and quantitative PCR were used to detect Dlk1 KO efficiency. Flow cytometry was performed to investigate the effects of Dlk1 KO on HSCs, progenitors and linage cells in primary mice. Competitive HSCs transplantation and secondary transplantation was used to examine the effects of Dlk1 KO on long-term hematopoietic repopulation potential of HSCs. RNA-Seq and cell metabolism assays was used to determine the underlying mechanisms. RESULTS: Dlk1 was highly expressed in adult mice long-term HSCs (LT-HSCs) relative to progenitors and mature lineage cells. Dlk1 KO in adult mice HSCs drove HSCs enter active cell cycle, and expanded phenotypical LT-HSCs, but undermined its long-term hematopoietic repopulation potential. Dlk1 KO resulted in an increase in HSCs' metabolic activity, including glucose uptake, ribosomal translation, mitochondrial metabolism and ROS production, which impaired HSCs function. Further, Dlk1 KO in adult mice HSCs attenuated Notch signaling, and re-activation of Notch signaling under Dlk1 KO decreased the mitochondrial activity and ROS production, and rescued the changes in frequency and absolute number of HSCs. Scavenging ROS by antioxidant N-acetylcysteine could inhibit mitochondrial metabolic activity, and rescue the changes in HSCs caused by Dlk1 KO. CONCLUSION: Our study showed that Dlk1 played an essential role in maintaining HSC homeostasis, which is realized by governing cell cycle and restricting mitochondrial metabolic activity.

Small but strong: Pivotal roles and potential applications of snoRNAs in hematopoietic malignancies.

Small nucleolar RNAs (snoRNAs) belong to a family of noncoding RNAs that are 60-300 nucleotides in length, and they are classified into two classes according to their structure and function: C/D box snoRNAs, playing an essential role in 2'-O-methylation modification on ribosomal RNA; H/ACA box snoRNAs, involved in the pseudouridylation of rRNA. SnoRNAs with unclear functions, no predictable targets, and unusual subcellular locations are called orphan snoRNAs. Recent studies have revealed abnormal expression and demonstrated the pivotal roles of snoRNAs and their host genes in various types of hematological malignancies. This review discusses recent discoveries concerning snoRNAs in a variety of hematological malignancies, including multiple myeloma, lymphoma and leukemia, and sheds light on the application of snoRNAs as diagnostic and prognostic markers as well as therapeutic targets of hematological malignancies in the future.

Observation and control of pseudospin switching in a finite-width topological photonic crystal.

Finite-size effect plays a significant role in topology photonics not to mention in reality all experimental setups are in finite-size. A photonic bandgap is opened in the topological edge state dispersion if a topological photonic crystal with finite width is considered, and the bandgap size relies on the finite-size effect. Pseudospin-preserving and pseudospin-flipping processes can be realized when a selectively switch of the pseudospin of edge states are customized by our designs. Our microwave experiments also successfully demonstrate pseudospin switch-on and -off behaviors in a finite-width photonic crystal. By combining photonic crystals with finite widths, a multi-tunneling proposal of topological photonic crystals can also be achieved. Our study of the finite-size effect will provide new approaches and thoughts to improve the development of topological photonic devices in the future.

UV and visible light photocatalytic activity of Au/TiO2 nanoforests with Anatase/Rutile phase junctions and controlled Au locations.

To magnify anatase/rutile phase junction effects through appropriate Au decorations, a facile solution-based approach was developed to synthesize Au/TiO2 nanoforests with controlled Au locations. The nanoforests cons®isted of anatase nanowires surrounded by radially grown rutile branches, on which Au nanoparticles were deposited with preferred locations controlled by simply altering the order of the fabrication step. The Au-decoration increased the photocatalytic activity under the illumination of either UV or visible light, because of the beneficial effects of either electron trapping or localized surface plasmon resonance (LSPR). Gold nanoparticles located preferably at the interface of anatase/rutile led to a further enhanced photocatalytic activity. The appropriate distributions of Au nanoparticles magnify the beneficial effects arising from the anatase/rutile phase junctions when illuminated by UV light. Under the visible light illumination, the LSPR effect followed by the consecutive electron transfer explains the enhanced photocatalysis. This study provides a facile route to control locations of gold nanoparticles in one-dimensional nanostructured arrays of multiple-phases semiconductors for achieving a further increased photocatalytic activity.