Sample preparation and data collection for serial block face scanning electron microscopy of mammalian cell monolayers.

Volume electron microscopy encompasses a set of electron microscopy techniques that can be used to examine the ultrastructure of biological tissues and cells in three dimensions. Two block face techniques, focused ion beam scanning electron microscopy (FIB-SEM) and serial block face scanning electron microscopy (SBF-SEM) have often been used to study biological tissue samples. More recently, these techniques have been adapted to in vitro tissue culture samples. Here we describe step-by-step protocols for two sample embedding methods for in vitro tissue culture cells intended to be studied using SBF-SEM. The first focuses on cell pellet embedding and the second on en face embedding. En face embedding can be combined with light microscopy, and this CLEM workflow can be used to identify specific biological events by light microscopy, which can then be imaged using SBF-SEM. We systematically outline the steps necessary to fix, stain, embed and image adherent tissue culture cell monolayers by SBF-SEM. In addition to sample preparation, we discuss optimization of parameters for data collection. We highlight the challenges and key steps of sample preparation, and the consideration of imaging variables.

Abundance, characteristics and ecological risks of microplastics from South Yellow Sea Mudflat.

Microplastic (MP) pollution in global marine environments has been extensively reported and attracted significant concerns, but MP distribution in mudflat has rarely been studied. In this paper, the abundance, features and ecological risk of MP in South Yellow Sea Mudflat were investigated comprehensively. MP were both detected in waters (5.4 ± 0.38-11.3 ± 0.78 items/L) and sediments (5.1 ± 0.36-10.1 ± 0.69 items/g) from South Yellow Sea Mudflat. There existed different MP abundance tendencies from sampling Group I (coastal estuary or port) and II (purely coastal mudflat), while MP abundance in water from Group II was lower than that from Group I generally, but MP abundance in sediment from Group I was lower than that from Group II generally. This suggested that MP abundance in mudflat water could be associated with frequent human activities significantly, and disturbance might not be beneficial to MP accumulation in sediments. Fragments, transparent, polyethylene (PE), polypropylene (PP) and polystyrene (PS) were major MP features in mudflat water and sediment, and maximum proportion of size of MP was 0.001-0.25 mm in both water and sediment. Furthermore, the primary risk assessment indicated that MP pollution load for mudflat was low level. However, potential MP ecological risk for mudflat could reach dangerous level to very dangerous level by calculating and evaluating polymer risk index (PRI) and potential ecological risk index (PERI), which could be caused by high proportions of polyvinyl chloride (PVC) and polyacrylonitrile (PAN) with high hazard score. For the first time, reference data about MP pollution from South Yellow Sea Mudflat were supplied in this paper, which would be helpful for management and control of MP in mudflat.

Central nervous injury risk factors after endovascular repair of a thoracic aortic aneurysm with type B aortic dissection.

BACKGROUND: Aortic dissection is the deadliest disease of the cardiovascular system. Type B aortic dissection accounts for 30%-60% of aortic dissections and is mainly treated by endovascular repair of thoracic endovascular aneurysm repair (TEVAR). However, patients are prone to various complications after surgery, with central nervous system injury being the most common, which seriously affects their prognosis and increases the risk of disability and death. Therefore, exploring the risk factors of central nervous system injury after TEVAR can provide a basis for its prevention and control. AIM: To investigate the risk factors for central nervous system injury after the repair of a thoracic endovascular aneurysm with type B aortic dissection. METHODS: We enrolled 306 patients with type B aortic dissection who underwent TEVAR at our hospital between December 2019 and October 2022. The patients were categorized into injury (n = 159) and non-injury (n = 147) groups based on central nervous system injury following surgery. The risk factors for central nervous system injury after TEVAR for type B aortic dissection were screened by comparing the two groups. Multivariate logistic regression analysis was performed. RESULTS: The Association between age, history of hypertension, blood pH value, surgery, mechanical ventilation, intensive care unit stay, postoperative recovery times on the first day after surgery, and arterial partial pressure of oxygen on the first day after surgery differed substantially (P < 0.05). Multivariate logistic regression analysis indicated that age, surgery time, history of hypertension, duration of mechanical ventilation, and intensive care unit stay were independent risk factors for central nervous system injury after TEVAR of type B aortic dissection (P < 0.05). CONCLUSION: For high-risk patients with central nervous system injury after TEVAR of type B aortic dissection, early intervention measures should be implemented to lower the risk of neurological discomfort following surgery in high-risk patients with central nervous system injury after TEVAR for type B aortic dissection.

Effects of Personality and Behavioral Syndromes on Competition for Social Hierarchical Status in Anemonefish Amphiprion clarkii.

In this study, the behavioral ethogram of Amphiprion clarkii during the growth phase prior to sexual differentiation was summarized based on behavioral observations in three social environments. These behaviors can be classified into four categories: in addition to normal behaviors, the other three categories of behaviors-threatening, agonistic, and appeasing behaviors-represent different intentions in interactions with other individuals. Subsequently, the personalities of each individual were assessed by testing their reactions to intruders. These individuals mainly exhibited two distinct personality types: bold-aggressive and shy-submissive. In pairing experiments, the interactive behaviors of the anemonefish were observed in pairing combinations of different body sizes and personalities. The impact of personality on the establishment of a stable social hierarchy was confirmed by significant differences in the success rates of different pairing combinations, with the frequency of appeasing behaviors being the main factor influencing the success rate. Our results suggested that in natural waters, when juvenile individuals migrate among host anemones, shy-submissive individuals are more likely to be accepted due to their appeasing behaviors towards larger individuals, thus avoiding the risk of being attacked and bitten, and benefiting the survival of the individual. Conversely, bold-aggressive individuals are more likely to be driven away to another host anemone due to their unwillingness to settle for a lower-ranked status, thereby contributing to population dispersal and increasing opportunities for gene exchange between populations.

LncRNA ERICD interacts with TROAP to regulate TGF-ß signaling in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most prevalent and common malignant tumors worldwide, accounting for 85-90 % of primary liver cancer cases. Accumulating evidence shows that long non-coding RNAs (LncRNAs) play regulatory roles in HCC occurrence and progression. However, little is known about the biological role of the LncRNA "E2F1-regulated inhibitor of cell death" (ERICD) in HCC. Our study revealed that ERICD is highly expressed in HCC and correlates with TNM staging; high ERICD levels were associated with poor patient prognoses. We revealed the targeting relationship between ERICD and miR-142-5p for the first time by bioinformatics prediction and further verified the targeting relationship between ERICD and miR-142-5p using a luciferase reporting experiment. In summary, our results showed that ERICD promotes the occurrence and metastasis of HCC by downregulating miR-142-5p expression. Our study provides a target for new potential therapeutic strategies for HCC.

Comparing pharmacotherapy and transcranial magnetic stimulation for the treatment of anxiety and depression after aortic dissection surgery.

BACKGROUND: Aortic coarctation is a potentially fatal condition that is primarily treated surgically. Despite successful procedures, patients frequently experience postoperative anxiety and depression, which can hinder recovery and worsen outcomes. Pharmacological interventions, such as 5-hydroxytryptamine (5-HT) and norepinephrine reuptake inhibitors, are commonly prescribed; however, their efficacy alone or in combination with non-invasive brain stimulation techniques, such as repetitive transcranial magnetic stimulation (TMS), remains unclear. AIM: To assess the effect of medications and TMS on post-aortic surgery anxiety and depression. METHODS: We analyzed the outcomes of 151 patients with anxiety and depression who were hospitalized for aortic dissection between January 2020 and September 2022. Using the random number table method, 75 and 76 patients were allocated to the normal control and study groups, respectively. All the patients were treated using routine procedures. The control group was administered anti-anxiety and anti-depression drugs, whereas the study group was treated with TMS in addition to these medications. The patients in both groups showed improvement after two courses of treatment. The Hamilton Anxiety Scale (HAMA) and the Hamilton Depression Scale (HAMD) were used to assess anxiety and depression, respectively. The serum levels of brain-derived neurotrophic factor (BDNF) and 5-HT were determined using enzyme-linked immunosorbent assay. The Pittsburgh Sleep Quality Index (PSQI) was used to estimate sleep quality, and the Repeatable Battery for the Assessment of Neuropsychological Status (RBANS) was used to assess cognitive function. RESULTS: The HAMD and HAMA scores reduced in 2 groups, with the study group achieving a lower level than control (P < 0.05). In the control group, 43 patients recovered, 17 showed improvement, and 15 were deemed invalid. In the study group, 52 recovered, 20 improved, and four were invalid. The efficacy rate in study group was 94.74% compared to 80.00% in control (P < 0.05). The BDNF and 5-HT levels increased in both groups, with higher levels observed in the experimental group (P < 0.05). Moreover, the PSQI scores decreased in 2 groups, but were lower in the intervention group than control (P < 0.05). The scores of the RBANS items increased, with the study group scoring higher than control (P < 0.05). CONCLUSION: Combining anti-anxiety and anti-depressive drugs with repetitive TMS after aortic surgery may enhance mood and treatment outcomes, offering a promising clinical approach.

Unveiling the Correlation Between the Membrane Assembly of P-gp and Drug Resistance in Multiple Myeloma Using Super-Resolution Fluorescence Imaging.

Drug resistance in multiple myeloma (MM) poses a significant challenge to treatment efficacy, primarily attributed to P-glycoprotein (P-gp) dysfunction. This study delves into the elusive spatial organization of P-gp, aiming to enhance our understanding of its role in MM drug resistance by exploring the intricate relationship between molecular function and spatial arrangement. Employing super-resolution imaging of P-gp with the inhibitor probe Tariquidar-TAMR labeling on MM cell membranes, the research uncovered a more pronounced clustering distribution of P-gp in drug-resistant cells (MM1R) compared to drug-sensitive counterparts (MM1S). Further exploration revealed the clustering distribution of P-gp was heightened as cellular drug resistance increased in hypoxic condition, directly emphasizing the strong correlation between P-gp cluster morphology and drug resistance. Additionally, stable P-gp cluster formation was influenced by cross-linking of membrane carbohydrates, and disrupting these glycoprotein clusters could reduce cellular drug resistance, suggesting that altering distribution patterns of P-gp can modulate drug responsiveness. Finally, dexamethasone (Dex) treatment was revealed to enhance P-gp clustering distribution, particularly in MM1S cells, indicating that change degree in P-gp distribution correlate with the modifiable space of cellular drug responsiveness. This study provides insights into the correlation between P-gp assembly and cellular drug responsiveness, deepening our understanding of functional changes in MM drug resistance and offering valuable perspectives for overcoming this challenge.

Diffusion-based culture and real-time impedance monitoring of tumor spheroids in hydrogel microwells of a suspended membrane under microfluidic conditions.

Tumor spheroids are widely studied for in vitro modeling of tumor growth and responses to anticancer drugs. However, current methods are mostly limited to static and perfusion-based cultures, which can be improved by more accurately mimicking pathological conditions. Here, we developed a diffusion-based dynamic culture system for tumor spheroids studies using a thin membrane of hydrogel microwells and a microfluidic device. This allows for effective exchange of nutrients and metabolites between the tumors and the culture medium flowing underneath, resulting in uniform tumor spheroids. To monitor the growth and drug response of the spheroids in real-time, we performed spectroscopic analyses of the system's impedance, demonstrating a close correlation between the tumor size and the resistance and capacitance of the system. Our results also indicate an enhanced drug effect on the tumor spheroids in the presence of a low AC electric field, suggesting a weakening mechanism of the spheroids induced by external perturbation.

Radiation-Induced Endothelial Ferroptosis Accelerates Atherosclerosis via the DDHD2-Mediated Nrf2/GPX4 Pathway.

This study sought to explore potential roles of endothelial ferroptosis in radiation-associated atherosclerosis (RAA) and molecular mechanisms behind this phenomenon. Here, an in vivo RAA mouse model was used and treated with ferroptosis inhibitors. We found that the RAA group had a higher plaque burden and a reduction in endothelial cells with increased lipid peroxidation compared to the control group, while ameliorated by liproxstatin-1. In vitro experiments further confirmed that radiation induced the occurrence of ferroptosis in human artery endothelial cells (HAECs). Then, proteomics analysis of HAECs identified domain-containing protein 2 (DDHD2) as a co-differentially expressed protein, which was enriched in the lipid metabolism pathway. In addition, the level of lipid peroxidation was elevated in DDHD2-knockdown HAECs. Mechanistically, a significant decrease in the protein and mRNA expression of glutathione peroxidase 4 (GPX4) was observed in HAECs following DDHD2 knockdown. Co-immunoprecipitation assays indicated a potential interaction between DDHD2 and nuclear factor erythroid 2-related factor 2 (Nrf2). The downregulation of Nrf2 protein was also detected in DDHD2-knockdown HAECs. In conclusion, our findings suggest that radiation-induced endothelial ferroptosis accelerates atherosclerosis, and DDHD2 is a potential regulatory protein in radiation-induced endothelial ferroptosis through the Nrf2/GPX4 pathway.

Fast-BEV: A Fast and Strong Bird's-Eye View Perception Baseline.

Recently, perception task based on Bird's-Eye View (BEV) representation has drawn more and more attention, and BEV representation is promising as the foundation for next-generation Autonomous Vehicle (AV) perception. However, most existing BEV solutions either require considerable resources to execute on-vehicle inference or suffer from modest performance. This paper proposes a simple yet effective framework, termed Fast-BEV, which is capable of performing faster BEV perception on the on-vehicle chips. Towards this goal, we first empirically find that the BEV representation can be sufficiently powerful without expensive transformer based transformation nor depth representation. Our Fast-BEV consists of five parts, We innovatively propose (1) a lightweight deploymentfriendly view transformation which fast transfers 2D image feature to 3D voxel space, (2) an multi-scale image encoder which leverages multi-scale information for better performance, (3) an efficient BEV encoder which is particularly designed to speed up on-vehicle inference. We further introduce (4) a strong data augmentation strategy for both image and BEV space to avoid over-fitting, (5) a multiframe feature fusion mechanism to leverage the temporal information. Among them, (1) and (3) enable Fast-BEV to be fast inference and deployment friendly on the on-vehicle chips, (2), (4) and (5) ensure that Fast-BEV has competitive performance. All these make Fast-BEV a solution with high performance, fast inference speed, and deployment-friendly on the on-vehicle chips of autonomous driving. Through experiments, on 2080Ti platform, our R50 model can run 52.6 FPS with 47.3% NDS on the nuScenes validation set, exceeding the 41.3 FPS and 47.5% NDS of the BEVDepth-R50 model [1] and 30.2 FPS and 45.7% NDS of the BEVDet4D-R50 model [2]. Our largest model (R101@900x1600) establishes a competitive 53.5% NDS on the nuScenes validation set. We further develop a benchmark with considerable accuracy and efficiency on current popular on-vehicle chips. The code is released at: https://github.com/Sense-GVT/FastBEV.

Assessing elevated pressure impact on photoelectrochemical water splitting via multiphysics modeling.

Photoelectrochemical (PEC) water splitting is a promising approach for sustainable hydrogen production. Previous studies have focused on devices operated at atmospheric pressure, although most applications require hydrogen delivered at elevated pressure. Here, we address this critical gap by investigating the implications of operating PEC water splitting directly at elevated pressure. We evaluate the benefits and penalties associated with elevated pressure operation by developing a multiphysics model that incorporates empirical data and direct experimental observations. Our analysis reveals that the operating pressure influences bubble characteristics, product gas crossover, bubble-induced optical losses, and concentration overpotential, which are crucial for the overall device performance. We identify an optimum pressure range of 6-8 bar for minimizing losses and achieving efficient PEC water splitting. This finding provides valuable insights for the design and practical implementation of PEC water splitting devices, and the approach can be extended to other gas-producing (photo)electrochemical systems. Overall, our study demonstrates the importance of elevated pressure in PEC water splitting, enhancing the efficiency and applicability of green hydrogen generation.

RhoGEF Tiam2 Regulates Glutamatergic Synaptic Transmission in Hippocampal CA1 Pyramidal Neurons.

Glutamatergic synapses exhibit significant molecular diversity, but circuit-specific mechanisms that underlie synaptic regulation are not well characterized. Prior reports show that Rho-guanine nucleotide exchange factor (RhoGEF) Tiam1 regulates perforant path→dentate gyrus granule neuron synapses. In the present study, we report Tiam1's homolog Tiam2 is implicated in glutamatergic neurotransmission in CA1 pyramidal neurons. We find that Tiam2 regulates evoked excitatory glutamatergic currents via a postsynaptic mechanism mediated by the catalytic Dbl-homology domain. Overall, we present evidence for RhoGEF Tiam2's role in glutamatergic synapse function at Schaffer collateral→CA1 pyramidal neuron synapses.

Regulating the vacancies of nitrogen-doped carbon nanohorns for high-performance electromagnetic wave absorption.

Single-walled carbon nanohorns (SWCNHs), which are sealed on one side with a conical cap and can self-aggregate, are aggregates with spherical morphology ranging from 30 to 100 nm and include dahlia, bud, and seed structures. These SWCNHs are suitable for electromagnetic wave absorption (EMWA) due to their conductivity loss. However, conductivity loss, which is part of three primary loss mechanisms, leads to SWCNHs suffering from impedance mismatching and a narrow effective absorption bandwidth (EAB). In this work, the content of vacancy-type defects in "dahlia-like" nitrogen-doped single-walled carbon nanohorns (NSWCNHs) is regulated by dielectric barrier discharge (DBD) plasma with argon to adjust their polarization and impedance matching. The high-energy argon ions from the plasma impact the bonds between the carbon atoms and adsorbed oxygen, leading to the sputtering of oxygen atoms from the surface and resulting in an increase in surface disorder and defect content. Vacancy-type defects improved polarization loss and optimized impedance matching, leading to the satisfactory EMWA performance of NSWCNHs. The NSWCNHs exhibit an outstanding minimum reflection loss (RLmin) of -57.94 dB when subjected to argon DBD treatment for 5 minutes, achieving this remarkable result at a thickness of 1.9 mm. Additionally, the effective absorption bandwidth (EAB) can cover 4.78 GHz after a treatment period of 1 minute. These results suggest that NSWCNHs have great potential as high-efficiency EMWA materials and demonstrate a new approach for designing high-performance EMWA absorbers.

Integrated PBPK-EO modeling of osimertinib to predict plasma concentrations and intracranial EGFR engagement in patients with brain metastases.

The purpose of this study was to develop and validate a physiologically based pharmacokinetic (PBPK) model combined with an EGFR occupancy (EO) model for osimertinib (OSI) to predict plasma trough concentration (Ctrough) and the intracranial time-course of EGFR (T790M and L858R mutants) engagement in patient populations. The PBPK model was also used to investigate the key factors affecting OSI pharmacokinetics (PK) and intracranial EGFR engagement, analyze resistance to the target mutation C797S, and determine optimal dosing regimens when used alone and in drug-drug interactions (DDIs). A population PBPK-EO model of OSI was developed using physicochemical, biochemical, binding kinetic, and physiological properties, and then validated using nine clinical PK studies, observed EO study, and two clinical DDI studies. The PBPK-EO model demonstrated good consistency with observed data, with most prediction-to-observation ratios falling within the range of 0.7 to 1.3 for plasma AUC, Cmax, Ctrough and intracranial free concentration. The simulated time-course of C797S occupancy by the PBPK model was much lower than T790M and L858R occupancy, providing an explanation for OSI on-target resistance to the C797S mutation. The PBPK model identified ABCB1 CLint,u, albumin level, and EGFR expression as key factors affecting plasma Ctrough and intracranial EO for OSI. Additionally, PBPK-EO simulations indicated that the optimal dosing regimen for OSI in patients with brain metastases is either 80 mg once daily (OD) or 160 mg OD, or 40 mg or 80 mg twice daily (BID). When used concomitantly with CYP enzyme perpetrators, the PBPK-EO model suggested appropriate dosing regimens of 80 mg OD with fluvoxamine (FLUV) itraconazole (ITR) or fluvoxamine (FLUC) for co-administration and an increase to 160 mg OD with rifampicin (RIF) or efavirenz (EFA). In conclusion, the PBPK-EO model has been shown to be capable of simulating the pharmacokinetic concentration-time profiles and the time-course of EGFR engagement for OSI, as well as determining the optimum dosing in various clinical situations.

Recent advances in the reciprocal regulation of m6A modification with non-coding RNAs and its therapeutic application in acute myeloid leukemia.

N6-methyladenosine (m6A) is one of the most common modifications of RNA in eukaryotic cells and is involved in mRNA metabolism, including stability, translation, maturation, splicing, and export. m6A also participates in the modification of multiple types of non-coding RNAs, such as microRNAs, long non-coding RNAs, and circular RNAs, thereby affecting their metabolism and functions. Increasing evidence has revealed that m6A regulators, such as writers, erasers, and readers, perform m6A-dependent modification of ncRNAs, thus affecting cancer progression. Moreover, ncRNAs modulate m6A regulators to affect cancer development and progression. In this review, we summarize recent advances in understanding m6A modification and ncRNAs and provide insights into the interaction between m6A modification and ncRNAs in cancer. We also discuss the potential clinical applications of the mechanisms underlying the interplay between m6A modifications and ncRNAs in acute myeloid leukemia (AML). Therefore, clarifying the mutual regulation between m6A modifications and ncRNAs is of great significance to identify novel therapeutic targets for AML and has great clinical application prospects.

Ultra-high pressure treatment improve the content of characteristic aromatic components of melon juice from the view of physical changes.

Introduction: The effectiveness of ultra-high pressure (UHP) technology in retaining the flavor of fresh fruit and vegetable juices has been acknowledged in recent years. Along with previously hypothesized conclusions, the improvement in melon juice flavor may be linked to the reduction of its surface tension through UHP. Methods: In this paper, the particle size, free-water percentage, and related thermodynamic parameters of melon juice were evaluated in a physical point for a deeper insight. Results: The results showed that the UHP treatment of P2-2 (200 MPa for 20 min) raised the free water percentage by 7,000 times than the other treatments and both the melting enthalpy, binding constant and Gibbs free energy of P2-2 were minimized. This significantly increased the volatility of characteristic aromatic compounds in melon juice, resulting in a 1.2-5 times increase in the content of aromatic compounds in the gas phase of the P2-2 group compared to fresh melon juice.

Mapping cell diversity in human sporadic cerebral cavernous malformations.

BACKGROUND: Cerebral cavernous malformation (CCM) is a low-flow, bleeding-prone vascular disease that can cause cerebral hemorrhage, seizure and neurological deficits. Its inheritance mode includes sporadic or autosomal dominant inheritance with incomplete penetrance, namely sporadic CCM (SCCM) and familial CCM. SCCM is featured by single lesion and single affection in a family. Among CCM patients especially SCCM, the pathogenesis of the corresponding phenotypes and pathological features or candidate genes have not been fully elucidated yet. METHODS: Here, we performed in-depth single-cell RNA sequencing (scRNA-Seq) and bulk assay for transposase-accessible chromatin sequencing (ATAC-Seq) in SCCM and control patients. Further validation was conducted for the gene of interest using qPCR and RNA in situ hybridization (RNA FISH) techniques to provide further atlas and evidence for SCCM generative process. RESULTS: We identified six cell types in the SCCM and control vessels and found that the expression of NEK1, RNPC3, FBRSL1, IQGAP2, MCUB, AP3B1, ESCO1, MYO9B and PVT1 were up-regulated in SCCM tissues. Among the six cell types, we found that compared with control conditions, PVT1 showed a rising peak which followed the pseudo-time axis in endothelial cell clusters of SCCM samples, while showed an increasing trend in smooth muscle cell clusters of SCCM samples. Further experiments indicated that, compared with the control vessels, PVT1 exhibited significantly elevated expression in SCCM samples. CONCLUSION: In SCCM conditions, We found that in the process of development from control to lesion conditions, PVT1 showed a rising peak in endothelial cells and showed an increasing trend in smooth muscle cells at the same time. Overall, there was a significantly elevated expression of NEK1, RNPC3, FBRSL1, IQGAP2, MCUB, AP3B1, ESCO1, MYO9B and PVT1 in SCCM specimens compared to control samples.

Integrative multi-omics profiling in human decedents receiving pig heart xenografts.

In a previous study, heart xenografts from 10-gene-edited pigs transplanted into two human decedents did not show evidence of acute-onset cellular- or antibody-mediated rejection. Here, to better understand the detailed molecular landscape following xenotransplantation, we carried out bulk and single-cell transcriptomics, lipidomics, proteomics and metabolomics on blood samples obtained from the transplanted decedents every 6 h, as well as histological and transcriptomic tissue profiling. We observed substantial early immune responses in peripheral blood mononuclear cells and xenograft tissue obtained from decedent 1 (male), associated with downstream T cell and natural killer cell activity. Longitudinal analyses indicated the presence of ischemia reperfusion injury, exacerbated by inadequate immunosuppression of T cells, consistent with previous findings of perioperative cardiac xenograft dysfunction in pig-to-nonhuman primate studies. Moreover, at 42 h after transplantation, substantial alterations in cellular metabolism and liver-damage pathways occurred, correlating with profound organ-wide physiological dysfunction. By contrast, relatively minor changes in RNA, protein, lipid and metabolism profiles were observed in decedent 2 (female) as compared to decedent 1. Overall, these multi-omics analyses delineate distinct responses to cardiac xenotransplantation in the two human decedents and reveal new insights into early molecular and immune responses after xenotransplantation. These findings may aid in the development of targeted therapeutic approaches to limit ischemia reperfusion injury-related phenotypes and improve outcomes.

Elucidating the Functional Mechanism of PTK7 in Cancer Development through Spatial Assembly Analysis Using Super Resolution Imaging.

Protein tyrosine kinase-7 (PTK7) has been reported as a vital participant in the Wnt signaling pathway, influencing tumorigenesis and metastasis. However, their specific roles in the mechanisms underlying cancer development and progression remain elusive. Here, using direct stochastic optical reconstruction microscopy (dSTORM) with aptamer-probe labeling, we first revealed that a weakening clustering distribution of PTK7 on the basal membranes happened as cellular migration increased during cancer progression. This correspondence was further supported by a diminished aggregated state of PTK7 caused by direct enhancement of cell migration. By comparing the alterations in PTK7 distribution with activation or inhibition of specific Wnt signaling pathway, we speculated that PTK7 could modulate cell migration by participating in the interplay between canonical Wnt (in MCF7 cells) and noncanonical Wnt signals (in MDA-MB-231 cells). Furthermore, we discovered that the spatial distribution morphology of PTK7 was also subject to the hydrolysis ability and activation state of the related hydrolase Matrix metallopeptidase14 (MMP14). This function-related specific assembly of PTK7 reveals a clear relationship between PTK7 and cancer. Meanwhile, potential molecular interactions predicted by the apparent assembly morphology can promote a deep understanding of the functional mechanism of PTK7 in cancer progress.

Minnelide exhibits antileukemic activity by targeting the Ars2/miR-190a-3p axis.

BACKGROUND: The identification of a novel and effective strategy for the clinical treatment of acute leukemia (AL) is a long-term goal. Minnelide, a water-soluble prodrug of triptolide, has recently been evaluated in phase I and II clinical trials in patients with multiple cancers and has shown promise as an antileukemic agent. However, the molecular mechanism underlying minnelide's antileukemic activity remains unclear. PURPOSE: To explore the molecular mechanisms by which minnelide exhibits antileukemic activity. METHODS: AL cells, primary human leukemia cells, and a xenograft mouse model were treated with triptolide and minnelide. The molecular mechanism was elucidated using western blotting, immunoprecipitation, flow cytometry, GSEA and liquid chromatography-mass spectrometry analysis. RESULTS: Minnelide was highly effective in inhibiting leukemogenesis and improving survival in two complementary AL mouse models. Triptolide, an active form of minnelide, causes cell cycle arrest in G1 phase and induces apoptosis in both human AL cell lines and primary AL cells. Mechanistically, we identified Ars2 as a new chemotherapeutic target of minnelide for AL treatment. We found that triptolide directly targeted Ars2, resulting in the downregulation of miR-190a-3p, which led to the disturbance of PTEN/Akt signaling and culminated in G1 cell cycle arrest and apoptosis. CONCLUSIONS: Our findings demonstrate that targeting Ars2/miR-190a-3p signaling using minnelide could represent a novel chemotherapeutic strategy for AL treatment and support the evaluation of minnelide for the treatment of AL in clinical trials.