Bulk T-cell receptor sequencing confirms clonality in obstetric antiphospholipid syndrome and may as a potential biomarker.

The heterogeneity of the T cell receptor (TCR) repertoire critically influences the autoimmune response in obstetric antiphospholipid syndrome (OAPS) and is intimately associated with the prophylaxis of autoimmune disorders. Investigating the TCR diversity patterns in patients with OAPS is thus of paramount clinical importance. This investigation procured peripheral blood specimens from 31 individuals with OAPS, 21 patients diagnosed with systemic lupus erythematosus (SLE), and 22 healthy controls (HC), proceeding with TCR repertoire sequencing. Concurrently, adverse pregnancy outcomes in the OAPS cohort were monitored and documented over an 18-month timeframe. We paid particular attention to disparities in V/J gene utilisation and the prevalence of shared clonotypes amongst OAPS patients and the comparative groups. When juxtaposed with observations from healthy controls and SLE patients, immune repertoire sequencing disclosed irregular T- and B-cell profiles and a contraction of diversity within the OAPS group. Marked variances were found in the genomic rearrangements of the V gene, J gene, and V/J combinations. Utilising a specialised TCRß repertoire, we crafted a predictive model for OAPS classification with robust discriminative capability (AUC = 0.852). Our research unveils alterations in the TCR repertoire among OAPS patients for the first time, positing potential covert autoimmune underpinnings. These findings nominate the TCR repertoire as a prospective peripheral blood biomarker for the clinical diagnosis of OAPS and may offer valuable insights for advancing the understanding of OAPS immunologic mechanisms and prognostic outcomes.

YAP inhibition overcomes adaptive resistance in HER2-positive gastric cancer treated with trastuzumab via the AKT/mTOR and ERK/mTOR axis.

BACKGROUND: Human epidermal growth factor receptor 2 (HER2)-positive gastric cancer (GC) is a heterogeneous GC subtype characterized by the overexpression of HER2. To date, few specific targeted therapies have demonstrated durable efficacy in HER2-positive GC patients, with resistance to trastuzumab typically emerging within 1 year. However, the mechanisms of resistance to trastuzumab remain incompletely understood, presenting a significant challenge to clinical practice. METHODS: In this study, we integrated genetic screening and bulk transcriptome and epigenomic profiling to define the mechanisms mediating adaptive resistance to HER2 inhibitors and identify potential effective therapeutic strategies for treating HER2-positive GCs. RESULTS: We revealed a potential association between adaptive resistance to trastuzumab in HER2-positive GC and the expression of YES-associated protein (YAP). Notably, our investigation revealed that long-term administration of trastuzumab triggers extensive chromatin remodeling and initiates YAP gene transcription in HER2-positive cells characterized by the initial inhibition and subsequent reactivation. Furthermore, treatment of HER2-positive GC cells and cell line-derived xenografts (CDX) models with YAP inhibitors in combination with trastuzumab was found to induce synergistic effects through the AKT/mTOR and ERK/mTOR pathways. CONCLUSION: These findings underscore the pivotal role of reactivated YAP and mTOR signaling pathways in the development of adaptive resistance to trastuzumab and may serve as a promising joint target to overcome resistance to trastuzumab.

Histone H3K18 and Ezrin Lactylation Promote Renal Dysfunction in Sepsis-Associated Acute Kidney Injury.

Histone lactylation is a metabolic stress-related histone modification. However, the role of histone lactylation in the development of sepsis-associated acute kidney injury (SA-AKI) remains unclear. Here, histone H3K18 lactylation (H3K18la) is elevated in SA-AKI, which is reported in this study. Furthermore, this lactate-dependent histone modification is enriched at the promoter of Ras homolog gene family member A (RhoA) and positively correlated with the transcription. Correction of abnormal lactate levels resulted in a reversal of abnormal histone lactylation at the promoter of RhoA. Examination of related mechanism revealed that histone lactylation promoted the RhoA/Rho-associated protein kinase (ROCK) /Ezrin signaling, the activation of nuclear factor-κB (NF-κB), inflammation, cell apoptosis, and aggravated renal dysfunction. In addition, Ezrin can undergo lactylation modification. Multiple lactylation sites are identified in Ezrin and confirmed that lactylation mainly occurred at the K263 site. The role of histone lactylation is revealed in SA-AKI and reportes a novel post-translational modification in Ezrin. Its potential role in regulating inflammatory metabolic adaptation of renal proximal tubule epithelial cells is also elucidated. The results provide novel insights into the epigenetic regulation of the onset of SA-AKI.

H3K4me3-Mediated FOXJ2/SLAMF8 Axis Aggravates Thrombosis and Inflammation in ß2GPI/Anti-ß2GPI-Treated Monocytes.

Antiphospholipid syndrome (APS) is characterized by thrombus formation, poor pregnancy outcomes, and a proinflammatory response. H3K4me3-related monocytes activation are key regulators of APS pathogenesis. Therefore, H3K4me3 CUT&Tag and ATAC-seq are performed to examine the epigenetic profiles. The results indicate that the H3K4me3 signal and chromatin accessibility at the FOXJ2 promoter are enhanced in an in vitro monocyte model by stimulation with ß2GPI/anti-ß2GPI, which mimics APS, and decreases after OICR-9429 administration. Furthermore, FOXJ2 is highly expressed in patients with primary APS (PAPS) and is the highest in patients with triple-positive antiphospholipid antibodies (aPLs). Mechanistically, FOXJ2 directly binds to the SLAMF8 promoter and activates SLAMF8 transcription. SLAMF8 further interacts with TREM1 to stimulate TLR4/NF-κB signaling and prohibit autophagy. Knockdown of FOXJ2, SLAMF8, or TREM1 blocks TLR4/NF-κB and provokes autophagy, subsequently inhibiting the release of inflammatory and thrombotic indicators. A mouse model of vascular APS is established via ß2GPI intraperitoneal injection, and the results suggest that OICR-9429 administration attenuates the inflammatory response and thrombus formation by inactivating FOXJ2/SLAMF8/TREM1 signaling. These findings highlight the overexpression of H3K4me3-mediated FOXJ2 in APS, which consequently accelerates APS pathogenesis by triggering inflammation and thrombosis via boosting the SLAMF8/TREM1 axis. Therefore, OICR-9429 is a promising candidate drug for APS therapy.

[Hologram quantitative structure-activity relationship on the gas chromatographic retention index of plant essential oil constituents].

The gas chromatography retention index (RI) is an important parameter for the identification of different types of compounds in the field of chromatographic analysis; however, the experimental collection of RI values is a extremely cumbersome process. Thus, there is an urgent need for the establishment of a simple, efficient, and accurate model for the prediction of the RI values of compounds. In this study, first, the experimental RI values for 60 plant essential oil constituents were obtained. Next, a model describing the hologram quantitative structure-activity relationship (HQSAR) between the structural properties of the essential oil constituents and their RI values was investigated and constructed. The optimal HQSAR model was established by setting the model parameters "fragment size", "fragment distinction", "hologram length" and "principal components" to "1-4", "C, Ch", "199", and "4", respectively. Finally, the predictive ability of the model was verified using external test set validation and leave-one-out cross-validation (LOO-CV). The experimental results were as follows, the root mean square error of prediction (RMSEP), predictive determination coefficient ([Formula: see text]), concordance correlation coefficient (CCC), and mean relative error (MRE) for external test set validation were 40.45, 0.984, 0.968, and 2.20%, respectively. Meanwhile, the root mean square error of cross validation (RMSECV) and MRE for LOO-CV were 72.56 and 4.17%, respectively. These findings demonstrate that the established HQSAR model has a good predictive ability and can accurately predict the RI values of plant essential oil constituents. In addition, the molecular contribution maps of the HQSAR model revealed that the RI values of aromatic compounds increase when hydroxyl groups are connected to their alkyl chains. Aliphatic compounds feature long chain alkyl groups, which can lead to an increase in RI values. The above phenomena highlight the promising application prospects of HQSAR for studying the RI values of plant essential oil constituents. Therefore, this study provides a reliable theoretical basis for predicting the RI values of other essential oil constituents.

Archaea-Inspired Switchable Nanochannels for On-Demand Lithium Detection by pH Activation.

With the rapid development of the lithium ion battery industry, emerging lithium (Li) enrichment in nature has attracted ever-growing attention due to the biotoxicity of high Li levels. To date, fast lithium ion (Li+) detection remains urgent but is limited by the selectivity, sensitivity, and stability of conventional technologies based on passive response processes. In nature, archaeal plasma membrane ion exchangers (NCLX_Mj) exhibit Li+-gated multi/monovalent ion transport behavior, activated by different stimuli. Inspired by NCLX_Mj, we design a pH-controlled biomimetic Li+-responsive solid-state nanochannel system for on-demand Li+ detection using 2-(2-hydroxyphenyl)benzoxazole (HPBO) units as Li+ recognition groups. Pristine HPBO is not reactive to Li+, whereas negatively charged HPBO enables specific Li+ coordination under alkaline conditions to decrease the ion exchange capacity of nanochannels. On-demand Li+ detection is achieved by monitoring the decline in currents, thereby ensuring precise and stable Li+ recognition (>0.1 mM) in the toxic range of Li+ concentration (>1.5 mM) for human beings. This work provides a new approach to constructing Li+ detection nanodevices and has potential for applications of Li-related industries and medical services.

Circ DENND4C inhibits pyroptosis and alleviates ischemia-reperfusion acute kidney injury by exosomes secreted from human urine-derived stem cells.

Acute kidney injury (AKI) is a disease characterised by acute onset, high mortality, and poor prognosis, and is mainly caused by ischemia-reperfusion (I/R). Human urine-derived stem cells (USCs) exhibit antioxidant, anti-inflammatory, and anti-apoptotic cytoprotective effects. Previously, we found that exosomes from USCs had the ability to inhibit apoptosis and protect kidneys from I/R injury. This study aimed to investigate the role of USC-derived exosomes (USC-Exos) in reducing pyroptosis and alleviating I/R-AKI. Models of HK-2 cells hypoxia-reoxygenation (H/R) and I/R kidney injury was established in Sprague Dawley rats to simulate AKI in vitro and in vivo. USC-Exos were isolated using ultracentrifugation and identified via electron microscopy and western blotting. USC-Exos were co-cultured with HK-2 cells and injected into rats via the tail vein. The expression of pyroptosis-related molecules (GSDMD, caspase-1, and NLRP-3) was verified using PCR and western blotting. Changes in renal function were reflected in the serum creatinine, urea, and cystatin C levels. The degree of renal injury was determined using haematoxylin and eosin and immunohistochemical staining. The levels of IL-1ß and IL-18 were detected using enzyme-linked immunosorbent assay (ELISA) to verify the role of USC-Exos in pyroptosis. Differentially expressed circRNAs in I/R rat kidneys were screened by transcriptome sequencing, and a dual-luciferase experiment was used to verify the interaction between upstream and downstream molecules. Ischemia-reperfusion resulted in significantly impaired renal function and expression of pyroptosis molecules, and significantly increased concentrations of inflammatory factors. These effects were reversed by injecting USC-Exos. Circ DENND4C was the most significantly decreased circRNA in I/R rat renal tissue, and knock-down of circ DENND4C can aggravate AKI in vivo and in vitro. DAVID(http://david.abcc.ncifcrf.gov) website showed that miR 138-5p/FOXO3a is a potential downstream target of circ DENND4C. Knock-down of circ DENND4C in HK-2 cells resulted in increased expression of miR 138-5p and increased miR 138-5p can reverse the regulation of FOXO3a. Dual-luciferase assay verified the reverse interaction between circ DENND4C, miR 138-5p, and FOXO3a. Exosomes promote cell proliferation and inhibit the activation of NLR family pyrin domain containing 3 through the circ DENND4C/miR 138-5p/FOXO3a pathway, thereby reducing pyroptosis and AKI. Circ DENND4C may be a potential therapeutic target for AKI.

ARID5B-mediated LINC01128 epigenetically activated pyroptosis and apoptosis by promoting the formation of the BTF3/STAT3 complex in ß2GPI/anti-ß2GPI-treated monocytes.

BACKGROUND: Alterations of the trimethylation of histone 3 lysine 4 (H3K4me3) mark in monocytes are implicated in the development of autoimmune diseases. Therefore, the purpose of our study was to elucidate the role of H3K4me3-mediated epigenetics in the pathogenesis of antiphospholipid syndrome (APS). METHODS: H3K4me3 Cleavage Under Targets and Tagmentation and Assay for Transposase-Accessible Chromatin were performed to determine the epigenetic profiles. Luciferase reporter assay, RNA immunoprecipitation, RNA pull-down, co-immunoprecipitation and chromatin immunoprecipitation were performed for mechanistic studies. Transmission electron microscopy and propidium iodide staining confirmed cell pyroptosis. Primary monocytes from patients with primary APS (PAPS) and healthy donors were utilised to test the levels of key molecules. A mouse model mimicked APS was constructed with beta2-glycoprotein I (ß2GPI) injection. Blood velocity was detected using murine Doppler ultrasound. RESULTS: H3K4me3 signal and open chromatin at the ARID5B promoter were increased in an in vitro model of APS. The epigenetic factor ARID5B directly activated LINC01128 transcription at its promoter. LINC01128 promoted the formation of the BTF3/STAT3 complex to enhance STAT3 phosphorylation. Activated STAT3 interacted with the NLRP3 promoter and subsequently stimulated pyroptosis and apoptosis. ARID5B or BTF3 depletion compensated for LINC01128-induced pyroptosis and apoptosis by inhibiting STAT3 phosphorylation. In mice with APS, ß2GPI exposure elevated the levels of key proteins of pyroptosis and apoptosis pathways in bone marrow-derived monocytes, reduced the blood velocity of the ascending aorta, increased the thrombus size of the carotid artery, and promoted the release of interleukin (IL)-18, IL-1ß and tissue factor. Patients with PAPS had the high-expressed ARID5B and LINC01128, especially those with triple positivity for antiphospholipid antibodies. Moreover, there was a positive correlation between ARID5B and LINC01128 expression. CONCLUSION: This study indicated that ARID5B/LINC01128 was synergistically upregulated in APS, and they aggravated disease pathogenesis by enhancing the formation of the BTF3/STAT3 complex and boosting p-STAT3-mediated pyroptosis and apoptosis, thereby providing candidate therapeutic targets for APS. HIGHLIGHTS: The H3K4me3 mark and chromatin accessibility at the ARID5B promoter are increased in vitro model mimicked APS. ARID5B-mediated LINC01128 induces pyroptosis and apoptosis via p-STAT3 by binding to BTF3. ARID5B is high- expressed in patients with primary APS and positively correlated with LINC01128 expression. OICR-9429 treatment mitigates pyroptosis and related inflammation in vivo and in vitro models mimicked APS.

A Single-Tube 21-Color Multiparameter Flow Cytometry Improve the Sensitivity of B-ALL MRD Analysis.

BACKGROUND: Detection of minimal residual disease (MRD) by multiparameter flow cytometry (MFC) is a well-established risk stratification factor and therapeutic modification strategy in B acute lymphoblastic leukemia (B-ALL). However, current 8 color (8c)-MFC for MRD detection had the sensitivity of 0.01% with false negative or positive. Hence, a more sensitive and applicable MFC-MRD method is urgently needed. The aim of this study is to establish a single-tube 21c-MFC method to detect B-ALL MRD, evaluate its performance, and to investigate its preliminary clinical application. METHODS: We selected 21 markers to establish a single-tube 21c-MFC method. The repeatability and sensitivity of this method was validated by adding Nalm-6 cells to normal bone marrow. Samples from control group (n = 6), B-ALL group (n = 7) and complete remission (CR) group (n = 26) were detected by 21c- and 8c-MFC separately. The expression characteristics of these markers was analyzed in control and B-ALL group, and the consistency of 21c- and 8c-MFC in detecting MRD was compared. RESULTS: Repeatability of this method was 1.91% of CV and sensitivity was up to 0.005%. In control group, the expression of CD81, CD97, and CD200 gradually decreased and CD44, HLA-DR, CD73, and CD72 gradually increased with the maturation of normal B cells. In B-ALL group, CD73stro, CD81low, CD44stro, CD123stro, and CD58stro showed high-frequency expression. The consistency rate of 21c- and 8c-MFC in detecting MRD was 96%. CONCLUSIONS: A single-tube 21c-MFC method was established for MRD detection in B-ALL and had higher sensitivity than the 8c-MFC method.

Synthesis of Porous MgAl-LDH on a Micelle Template and Its Application for Efficient Treatment of Oilfield Wastewater.

In this paper, a series of porous hierarchical Mg/Al layered double hydroxides (named as LDH, TTAC-MgAl-LDH, CTAC-MgAl-LDH, and OTAC-MgAl-LDH) was synthesized by a simple green hydrothermal method using wormlike micelles formed by salicylic acid and surfactants with different carbon chain lengths (0, 14, 16, and 18) as soft templates. BET, XRD, FTIR, TG, and SEM characterizations were carried out in order to investigate the structure and properties of the prepared materials. The results showed that the porous hierarchical CTAC-MgAl-LDH had a large specific surface area and multiple pore size distributions which could effectively increase the reaction area and allow better absorption capability. Benefiting from the unique architecture, CTAC-MgAl-LDH exhibited a large adsorption capacity for sulfonated lignite (231.70 mg/g) at 25 °C and a pH of 7, which outperformed the traditional LDH (86.05 mg/g), TTAC-MgAl-LDH (108.15 mg/g), and OTAC-MgAl-LDH (110.51 mg/g). The adsorption process of sulfonated lignite followed the pseudo-second-order kinetics model and conformed the Freundlich isotherm model with spontaneous heat absorption, which revealed that electrostatic adsorption and ion exchange were the main mechanisms of action for the adsorption. In addition, CTAC-MgAl-LDH showed a satisfactory long-time stability and its adsorption capacities were still as high as 198.64 mg/g after two adsorption cycles.

Neuron-Inspired Nanofluidic Biosensors for Highly Sensitive and Selective Imidacloprid Detection.

Pesticides have caused concerns about food safety due to their residual effects in vegetables and fruits. Imidacloprid, as the frequently used neonicotinoid pesticide, could harm cardiovascular and respiratory function and cause reproductive toxicity in humans. Therefore, reliable methods for portable, selective, and rapid detection are desirable to develop. Herein, we report a neuron-inspired nanofluidic biosensor based on a tyrosine-modified artificial nanochannel for sensitively detecting imidacloprid. The functional tyrosine is modified on the outer surface of porous anodic aluminum oxide to rapidly capture imidacloprid through π-π interactions and hydrogen bonds. The integrated nanofluidic biosensor has a wide concentration range from 10-8 to 10-4 g/mL with an ultralow detection limit of 6.28 × 10-9 g/mL, which outperforms the state-of-the-art sensors. This work provides a new perspective on detecting imidacloprid residues as well as other hazardous pesticide residues in environmental and food samples.

Use of Highly Dispersed Mixed Metal Hydroxide Gel Compared to Bentonite Based Gel for Application in Drilling Fluid under Ultra-High Temperatures.

In order to solve the problem of poor dispersion and stability of mixed metal hydroxide (MMH), a kind of mixed metal hydroxide-like compound (MMHlc) gel was synthesized for use as the base mud in drilling fluid instead of bentonite gel. Na2CO3, Na2SiO3, and C17H33CO2Na were used as precipitants to form MMHlc with larger interlayer spacing and smaller particle size. MMHlc was synthesized by the coprecipitation method at 25 °C with a metal molar ratio of Mg:Al:Fe = 3:1:1. The performance evaluation of the treated drilling fluid showed that MMHlc (S2) synthesized using Na2SiO3 as the precipitant had the characteristics of low viscosity, low filtration, and a high dynamic plastic ratio at 25 °C, which fully met the requirements of oil field application, and it maintained its excellent properties after being aged at 250 °C for 16 h. Linear expansion and rolling recovery experiments showed that the S2 sample had excellent rheological properties and good inhibition. X-ray diffraction and FT-IR experiments showed that S2 had the most complete crystal structure, its interlayer distance was large, and its ion exchange capacity was strong. The thermogravimetric experiment showed that the S2 crystal was stable and the temperature resistance of the crystal could reach 340 °C. Zeta potential, particle size analysis, SEM, and TEM results showed that S2 is a nanomaterial with a complete morphology and uniform distribution. The drilling fluid of this formula had the characteristics of low viscosity, low filtration loss, and a high dynamic plastic ratio, and it met the conditions for oil field application. Considering these results, the new MMH prepared by our research institute is a drilling fluid material that can be used at ultra-high temperatures and can provide important support for drilling ultra-deep wells.

Construction of a hierarchical membrane with angstrom-scale ion channels for enhanced Li+/Mg2+ separation.

A biomimetic hierarchical membrane consisting of ZIF-8 and MXene with controllable morphology could be fabricated by the facile electrochemical deposition method, well-realizing Li+/Mg2+ sieving. This membrane could work stably in real brine with perm-selectivity of Li+/Mg2+ up to 47.4.

IGH rod-like tracer: An AlphaFold2 structural similarity extraction-based predictive biomarker for MRD monitoring in pre-B-ALL.

Sequence variation resulting from the evolution of IGH clones and immunophenotypic drift makes it difficult to track abnormal B cells in children with precursor B cell acute lymphoblastic leukemia (pre-B-ALL) by flow cytometry, qPCR, or next-generation sequencing (NGS). The V-(D)-J regions of immunoglobulin and T cell receptor of 47 pre-B-ALL samples were sequenced using the Illumina NovaSeq platform. The IGH rod-like tracer consensus sequence was extracted based on its rod-like alpha-helices structural similarity predicted by AlphaFold2. Additional data from published 203 pre-B-ALL samples were applied for validation. NGS-IGH (+) patients with pre-B-ALL had a poor prognosis. Consistent CDR3-coded protein structures in NGS-IGH (+) samples could be extracted as a potential follow-up marker for pre-B-ALL children during treatment. IGH rod-like tracer from quantitative immune repertoire sequencing may serve as a class of biomarker with significant predictive values for the dynamic monitoring of MRD in pre-B-ALL children.

Engineering Multi-field-coupled Synergistic Ion Transport System Based on the Heterogeneous Nanofluidic Membrane for High-Efficient Lithium Extraction.

The global carbon neutrality strategy brings a wave of rechargeable lithium-ion batteries technique development and induces an ever-growing consumption and demand for lithium (Li). Among all the Li exploitation, extracting Li from spent LIBs would be a strategic and perspective approach, especially with the low energy consumption and eco-friendly membrane separation method. However, current membrane separation systems mainly focus on monotonous membrane design and structure optimization, and rarely further consider the coordination of inherent structure and applied external field, resulting in limited ion transport. Here, we propose a heterogeneous nanofluidic membrane as a platform for coupling multi-external fields (i.e., light-induced heat, electrical, and concentration gradient fields) to construct the multi-field-coupled synergistic ion transport system (MSITS) for Li-ion extraction from spent LIBs. The Li flux of the MSITS reaches 367.4 mmol m-2 h-1, even higher than the sum flux of those applied individual fields, reflecting synergistic enhancement for ion transport of the multi-field-coupled effect. Benefiting from the adaptation of membrane structure and multi-external fields, the proposed system exhibits ultrahigh selectivity with a Li+/Co2+ factor of 216,412, outperforming previous reports. MSITS based on nanofluidic membrane proves to be a promising ion transport strategy, as it could accelerate ion transmembrane transport and alleviate the ion concentration polarization effect. This work demonstrated a collaborative system equipped with an optimized membrane for high-efficient Li extraction, providing an expanded strategy to investigate the other membrane-based applications of their common similarities in core concepts.

Efficient Solar-osmotic Power Generation from Bioinspired Anti-fouling 2D WS2 Composite Membranes.

Nanofluidic reverse electrodialysis provides an attractive way to harvest osmotic energy. However, most attention was paid to monotonous membrane structure optimization to promote selective ion transport, while the role of external fields and relevant mechanisms are rarely explored. Here, we demonstrate a Kevlar-toughened tungsten disulfide (WS2 ) composite membrane with bioinspired serosa-mimetic structures as an efficient osmotic energy generator coupling light. As a result, the output power could be up to 16.43 W m-2 under irradiation, outperforming traditional two-dimensional (2D) membranes. Both the experiment and simulation uncover that the generated photothermal and photoelectronic effects could synergistically promote the confined ion transport process. In addition, this membrane also possesses great anti-fouling properties, endowing its practical application. This work paves new avenues for sustainable power generation by coupling solar energy.

UBE4A catalyzes NRF1 ubiquitination and facilitates DDI2-mediated NRF1 cleavage.

The transcription factor nuclear factor erythroid 2 like 1 (NFE2L1 or NRF1) regulates constitutive and inducible expression of proteasome subunits and assembly chaperones. The precursor of NRF1 is integrated into the endoplasmic reticulum (ER) and can be retrotranslocated from the ER to the cytosol where it is processed by ubiquitin-directed endoprotease DDI2. DDI2 cleaves and activates NRF1 only when NRF1 is highly polyubiquitinated. It remains unclear how retrotranslocated NRF1 is primed with large amount of ubiquitin and/or very long polyubiquitin chain for subsequent processing. Here, we report that E3 ligase UBE4A catalyzes ubiquitination of retrotranslocated NRF1 and promotes its cleavage. Depletion of UBE4A reduces the amount of ubiquitin modified on NRF1, shortens the average length of polyubiquitin chain, decreases NRF1 cleavage efficiency and causes accumulation of non-cleaved, inactivated NRF1. Expression of a UBE4A mutant lacking ligase activity impairs the cleavage, likely due to a dominant negative effect. UBE4A interacts with NRF1 and the recombinant UBE4A can promote ubiquitination of retrotranslocated NRF1 in vitro. In addition, knocking out UBE4A reduces transcription of proteasomal subunits in cells. Our results indicate that UBE4A primes NRF1 for DDI2-mediated activation to facilitate expression of proteasomal genes.

Artificial intelligence of digital morphology analyzers improves the efficiency of manual leukocyte differentiation of peripheral blood.

BACKGROUND AND OBJECTIVE: Morphological identification of peripheral leukocytes is a complex and time-consuming task, having especially high requirements for personnel expertise. This study is to investigate the role of artificial intelligence (AI) in assisting the manual leukocyte differentiation of peripheral blood. METHODS: A total of 102 blood samples that triggered the review rules of hematology analyzers were enrolled. The peripheral blood smears were prepared and analyzed by Mindray MC-100i digital morphology analyzers. Two hundreds leukocytes were located and their cell images were collected. Two senior technologists labeled all cells to form standard answers. Afterward, the digital morphology analyzer unitized AI to pre-classify all cells. Ten junior and intermediate technologists were selected to review the cells with the AI pre-classification, yielding the AI-assisted classifications. Then the cell images were shuffled and re-classified without AI. The accuracy, sensitivity and specificity of the leukocyte differentiation with or without AI assistance were analyzed and compared. The time required for classification by each person was recorded. RESULTS: For junior technologists, the accuracy of normal and abnormal leukocyte differentiation increased by 4.79% and 15.16% with the assistance of AI. And for intermediate technologists, the accuracy increased by 7.40% and 14.54% for normal and abnormal leukocyte differentiation, respectively. The sensitivity and specificity also significantly increased with the help of AI. In addition, the average time for each individual to classify each blood smear was shortened by 215 s with AI. CONCLUSION: AI can assist laboratory technologists in the morphological differentiation of leukocytes. In particular, it can improve the sensitivity of abnormal leukocyte differentiation and lower the risk of missing detection of abnormal WBCs.

Antioxidant Carboxymethyl Chitosan Carbon Dots with Calcium Doping Achieve Ultra-Low Calcium Concentration for Iron-Induced Osteoporosis Treatment by Effectively Enhancing Calcium Bioavailability in Zebrafish.

Iron overloads osteoporosis mainly occurs to postmenopausal women and people requiring repeated blood transfusions. Iron overload increases the activity of osteoclasts and decreases the activity of osteoblasts, leading to the occurrence of osteoporosis. Conventional treatment options include calcium supplements and iron chelators. However, simple calcium supplementation is not effective, and it does not have a good therapeutic effect. Oxidative stress is one of the triggers for osteoporosis. Therefore, the study focuses on the antioxidant aspect of osteoporosis treatment. The present work revealed that antioxidant carboxymethyl chitosan-based carbon dots (AOCDs) can effectively treat iron overload osteoporosis. More interestingly, the functional modification of AOCDs by doping calcium gluconate (AOCDs:Ca) is superior to the use of any single component. AOCDs:Ca have the dual function of antioxidant and calcium supplement. AOCDs:Ca effectively improve the bioavailability of calcium and achieve ultra-low concentration calcium supplement for the treatment of iron-induced osteoporosis in zebrafish.

Multi-Mixed Metal Hydroxide as a Strong Stratigraphic Nanoclay Inhibitor in Solid-Free Drilling Fluid.

Solid-free drilling fluid has more advantages as a new type of drilling fluid compared with traditional drilling fluid, such as improving drilling efficiency, protecting oil and not having clay particles clog the oil and gas layer. In this study, Zn/Cu/Fe-doped magnesium-aluminum hydroxide (Mg-Al MMH) was prepared using the co-precipitation method and evaluated in solid-free drilling fluid. The inhibition mechanism of synthesized hydroxide was analyzed by X-ray diffraction, laser particle-size analysis and thermogravimetric analysis. The samples were directly used as drilling fluid base muds for performance evaluation. The results showed that the linear expansion rate of 4% M6-Fe was only 12.32% at room temperature within 2 h, that the linear expansion rate was 20.28% at 90 °C and that the anti-swelling rate was 81.16% at room temperature, indicating that it has a strong inhibition ability at both room temperature and at high temperatures. Meanwhile, the possibility of multi-mixed metal hydroxide as a drilling fluid base mud is discussed in this study. We found that 4% M6-Fe exhibited low viscosity, a high YP/PV ratio and high temperature resistance, and its apparent viscosity retention rate reached 100% rolled at 200 °C for 16 h, with a YP/PV ratio of 2.33.