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Abstract Background Patients with programmed cell death‐ligand 1 (PD‐L1) ≥50% metastatic non‐small cell lung cancer (NSCLC) treated with first‐line immunotherapy showed heterogeneous tumor responses. In this study, we investigated the clinical and immune‐inflammatory markers distinguishing patients with metastatic NSCLC achieving high depth of tumor response (HDPR) from those with non‐high depth of response (NHDPR). The impact of clinical features on the prognosis of patients with PD‐L1 ≥50% were further clarified. Methods The clinical characteristics and immune‐inflammatory markers of 17 patients with PD‐L1 ≥50% metastatic NSCLC at Beijing Tiantan Hospital between July 2020 and December 2023 were retrospectively analyzed. Results Among the 17 patients, seven (41.2%) patients achieved HDPR (range: −50%, −72%) and 10 (58.8%) patients achieved NHDPR (range: −13%, −45%). Below normal CD4 + T lymphocytes/CD8 + T lymphocytes (CD4/CD8) ratio (p = 0.01) and oncogenes and/or tumor suppressor gene mutations (TP53/KRAS/EGFR) (p = 0.001) were found enriched for NHDPR compared with HDPR. With a median follow‐up of 26.0 months (range: 17.2–34.8 months), the median progression‐free survival (PFS) following first‐line immunotherapy and overall survival (OS) were 9.0 months (95% CI: 5.0–13.0) and not reached (NR), respectively. The neutrophil‐to‐lymphocyte ratio (NLR) was identified as an independent prognostic factor on first‐line PFS. Patients with an NLR ≥4 exhibited a shorter median PFS (7.0 months vs. NR; p = 0.033; 95% CI: 1.2–80.2) than those with an NLR

Key Clinical Message Merkel cell carcinoma (MCC) is a rare and aggressive skin cancer that can be easily misdiagnosed in its early stages. Clinicians should maintain a high index of suspicion for rapidly growing skin lesions in elderly patients and promptly investigate with histopathology and immunohistochemistry. Treatment for MCC should be individualized, considering the patient's age, overall health, and quality of life. Close follow‐up is essential to detect recurrence or metastasis early.

BackgroundRegorafenib, a tyrosine kinase inhibitor (TKI), is used in the treatment of unresectable hepatocellular carcinoma (HCC). However, the occurrence of acquired resistance limits its antitumor efficacy. While multiple studies have highlighted the crucial role of bypass activation in acquired TKI resistance, few have focused on bypass activation in regorafenib resistance in HCC.MethodsHigh-throughput proteomics was used to identify differential proteins associated with bypass activation between acquired regorafenib-resistant cells and parental cells. The ability of epidermal growth factor receptor (EGFR) bypass inhibition to reverse resistance was evaluated both in vitro and in vivo using direct microscopic observation, the CCK-8 assay, colony formation assay, Annexin V-FITC/propidium iodide double staining, cell cycle analysis, western blotting, and a xenograft model.ResultsThe expression of EGFR, a member of the receptor tyrosine kinase (RTK) family, was significantly increased in acquired regorafenib-resistant HCC cells compared with parental cells. Pharmacological inhibition of EGFR with gefitinib restored the sensitivity of regorafenib-resistant HCC cells to regorafenib. In a xenograft mouse model, gefitinib sensitized resistant tumors to regorafenib. Additionally, levels of RAS, RAF, and P-ERK1/2, components of the downstream EGFR signaling pathway, were positively associated with EGFR expression.ConclusionEGFR overexpression promotes acquired resistance to regorafenib through RAS/RAF/ERK bypass activation in HCC. Inhibition of EGFR restores sensitivity to regorafenib, and the combination of gefitinib and regorafenib demonstrates significant antitumor efficacy both in vivo and in vitro. These findings suggest that this combination could be a potential strategy for patients with advanced HCC.

ABSTRACT Aspergillus fumigatus is the predominant pathogen responsible for aspergillosis infections, with emerging drug-resistant strains complicating treatment strategies. The role of mitochondrial functionality in fungal resistance to antifungal agents is well-documented yet not fully understood. In this study, the mitochondrial protein Bcs1A, a homolog of yeast Bcs1, was found to regulate colony growth, ion homeostasis, and the response to antifungal drugs in A. fumigatus. Microscopic observations revealed substantial colocalization of Bcs1A-GFP fusion protein fluorescence with mitochondria. Bcs1A deletion compromised colony growth and the utilization of non-fermentable carbon sources, alongside causing abnormal mitochondrial membrane potential and reduced reactive oxygen species production. These findings underscore Bcs1A’s vital role in maintaining mitochondrial integrity. Phenotypic analysis and determinations of minimum inhibitory concentrations indicated that the Δbcs1A mutant was more resistant to various antifungal agents, such as azoles, terbinafine, and simvastatin, compared to wild-type strain. RNA sequencing and RT-qPCR analysis highlighted an upregulation of multiple efflux pumps in the Δbcs1A mutant. Furthermore, loss of the principal drug efflux pump, mdr1, decreased azole tolerance in the Δbcs1A mutant, suggesting that Bcs1A’s modulated of azoles response via efflux pump expression. Collectively, these results establish Bcs1A as essential for growth and antifungal drug responsiveness in A. fumigatus mediated through mitochondrial regulation.IMPORTANCEDrug resistance presents a formidable obstacle in the clinical management of aspergillosis. Mitochondria are integral to various biochemical pathways, including those involved in fungi drug response, making mitochondrial proteins promising therapeutic targets for drug therapy. This study confirms that Bcs1A, a mitochondrial respiratory chain protein, is indispensable for mitochondrial functionality and multidrug tolerance in Aspergillus fumigatus. Mutation of Bcs1A not only leads to a series of drug efflux pumps upregulated but also shows that loss of the primary efflux pump, mdr1, partial reduction in drug tolerance in the Bcs1A mutant, highlighting that Bcs1A’s significant influence on mitochondria-mediated drug resistance.

The important role of cholesterol in tumor metastasis has been widely studied in recent years. Ezetimibe is currently the only selective cholesterol uptake inhibitor on the market. Here, we explored the effect of ezetimibe on breast cancer metastasis by studying its impact on breast cancer cell migration, invasion, and epithelial‐mesenchymal transition (EMT). Differential gene expression analysis and validation were also carried out to compare ezetimibe‐treated and untreated breast cancer cells. Finally, breast cancer cells overexpressing TGFβ2 were constructed, and the effect of TGFβ2 on the migration and invasion of ezetimibe‐treated breast cancer cells was examined. Our results show that ezetimibe treatment of breast cancer cells inhibited cell migration, invasion, and EMT, and it significantly suppressed the expression of TGFβ2. Overexpression of TGFβ2 reversed the inhibitory effect of ezetimibe on the migration and invasion of breast cancer cells. Taken together, our results suggest that ezetimibe might be a potential candidate for the treatment of breast cancer metastasis.

Key Clinical Message This case illustrates the clinical heterogeneity of porokeratosis (PK), with a patient presenting with both disseminated superficial actinic PK‐like facial lesions and PK of Mibelli‐like lesions on the buttocks and lower limbs. Ultraviolet exposure, infection, and immunosuppression may contribute to the manifestation of multiple clinical forms in a single patient. Close monitoring for potential malignant transformation is essential, particularly in elderly patients with long disease duration and a history of oncological conditions.

Numerous studies have highlighted the correlation between metal intake and deteriorated pulmonary function, emphasizing its pivotal role in the progression of Chronic Obstructive Pulmonary Disease (COPD). However, the efficacy of traditional models is often compromised due to overfitting and high bias in datasets with low-level exposure, rendering them ineffective in delineating the contemporary risk trends associated with pulmonary diseases. To address these limitations, we embarked on developing advanced, interpretable models, crucial for elucidating the intricate mechanisms of metal toxicity and enriching the domain knowledge embedded in toxicity models. In this endeavor, we scrutinized extensive, long-term metal exposure datasets from NHANES to explore the interplay between metal and pulmonary functionality. Employing a variety of machine-learning approaches, we opted for the “Mixer of Experts” model for its proficiency in identifying a myriad of toxicological trends and sensitivities. We conceptualized and illustrated the TSAP (Toxicity Score at Population-level), a metal interpretable scoring system offering performance nearly equivalent to the amalgamation of standard interpretable methods addressing the “black box” conundrum. This streamlined, bifurcated procedural analysis proved instrumental in discerning established risk factors, thereby uncovering Tungsten as a novel contributor to COPD risk. Synopsis: TSAP achieved satisfied performance with transparent interpretability, suggesting tungsten intake need further action for COPD prevention.

Background and Purpose: Regorafenib was approved by the US Food and Drug Administration (FDA) for hepatocellular carcinoma (HCC) patients showing progress on sorafenib treatment. However, there is an inevitably high rate of drug resistance associated with regorafenib, which reduces its effectiveness in clinical treatment. Thus, there is an urgent need to find a potential way to solve the problem of regorafenib resistance. The metabolite of disulfiram complexed with copper, the Diethyldithiocarbamate-copper complex (CuET), has been found to be an effective anticancer drug candidate. In the present study, we aimed to evaluate the effect of CuET on regorafenib resistance in HCC and uncover the associated mechanism. Experimental Approach: Regorafenib-resistant HCC strains were constructed by applying an increasing concentration gradient. This study employed a comprehensive range of methodologies, including the cell counting kit-8 (CCK-8) assay, colony formation assay, cell cycle analysis, wound healing assay, Transwell assay, tumor xenograft model, and immunohistochemical analysis. These methods were utilized to investigate the antitumor activity of CuET, assess the combined effect of regorafenib and CuET, and elucidate the molecular mechanism underlying CuET-mediated regorafenib resistance. Key Results: The inhibitory effect of regorafenib on cell survival, proliferation and migration was decreased in regorafenib-resistant MHCC-97H (MHCC-97H/REGO) cells compared with parental cells. CuET demonstrated significant inhibitory effects on cell survival, proliferation, and migration of various HCC cell lines. CuET restored the sensitivity of MHCC-97H/REGO HCC cells to regorafenib in vitro and in vivo. Mechanistically, CuET reverses regorafenib resistance in HCC by suppressing epithelial-mesenchymal transition (EMT) through inhibition of the ERK signaling pathway. Conclusion and Implications: Taken together, the results of this study demonstrated that CuET inhibited the activation of the ERK signaling pathway, leading to the suppression of the epithelial-mesenchymal transition (EMT) and subsequently reversing regorafenib resistance in HCC both in vivo and in vitro. This study provides a new idea and potential strategy to improve the treatment of regorafenib-resistant HCC.

Abstract Background Immunotherapy targeting PD-1/PD-L1 has revolutionized the treatment of extensive-stage small cell lung cancer (ES-SCLC). However, clinical trials suggest differential efficacy of anti-PD-1 agents and anti-PD-L1 agents in first-line treatment of ES-SCLC. This retrospective multicenter study aimed to compare the efficacy and safety of anti-PD-1 agents versus anti-PD-L1 agents in first-line treatment of ES-SCLC in real-world practice. Methods Patients with pathologically or cytologically confirmed ES-SCLC treated with platinum plus etoposide combined with anti-PD-1 or PD-L1 agents as first-line treatment in different centers of PLA General Hospital between January 2017 and October 2021 were included for this study. Survival outcomes and safety were compared between patients receiving anti-PD-1 and PD-L1 agents. Results Of the total 154 included patients, 68 received anti-PD-1 agents plus chemotherapy (PD-1 group), and 86 received anti-PD-L1 agents plus chemotherapy (PD-L1 group). Progression-free survival (PFS) and overall survival (OS) in the entire cohort were 7.6 months (95% confidence interval [CI]: 6.5–8.2 months) and 17.4 months (95% CI: 15.3–19.3 months), respectively. Median PFS and OS were comparable between the PD-1 group and PD-L1 group (PFS: 7.6 months vs. 8.3 months, HR = 1.13, 95% CI: 0.79–1.62, p = 0.415; OS: 26.9 months vs. 25.6 months, HR = 0.96, 95% CI: 0.63–1.47, p = 0.859. The objective response rate and disease control rate were comparable between the two groups: 79.4% vs. 79.1% and 92.6% vs. 94.2%, respectively. The 6-month, 12-month, and 18-month PFS and OS rates were slightly higher in the PD-L1 group than in the PD-1 group, while the 24-month PFS rate was slightly higher in the PD-1 group than in the PD-L1 group. Stratified analysis showed that locoregional thoracic radiotherapy and normal lactate dehydrogenase level were independent predictors of better OS in ES-SCLC patients treated with first-line chemotherapy plus ICI. Adverse events were not significantly different between the two groups. Conclusions Anti-PD-1 agents and anti-PD-L1 agents combined with chemotherapy as first-line treatment for ES-SCLC are comparably effective and well tolerated.

Cerebrospinal fluid-contacting neurons (CSF-cNs) represent a distinct group of interneurons characterized by their prominent apical globular protrusions penetrating the spinal cord’s central canal and their basal axons extending towards adjacent cells. Identified nearly a century back, the specific roles and attributes of CSF-cNs have just started to emerge due to the historical lack of definitive markers. Recent findings have confirmed that CSF-cNs expressing PKD2L1 possess attributes of neural stem cells, suggesting a critical function in the regeneration processes following spinal cord injuries. This review aims to elucidate the molecular markers of CSF-cNs as potential neural stem cells during spinal cord development and assess their roles post-spinal cord injury, with an emphasis on their potential therapeutic implications for spinal cord repair.

Abstract Circulating tumor DNA (ctDNA) has been widely used as a minimally invasive biomarker in clinical routine. However, a number of factors such as panel design, sample quality, patients' disease stages are known to influence ctDNA detection sensitivity. In this study, we systematically evaluated common factors associated with the variability of ctDNA detection in plasma and investigated ctDNA abundance in bronchoalveolar lavage (BAL). Whole exome profiling was conducted on 61 tumor tissue samples to identify tumor‐specific variants, which were then used to design personalized assay MarRyDa® for ctDNA detection. DNA extracted from BAL fluid and plasma were genotyped using MarRyDa® platform. Our analysis showed that histological subtypes and disease stages had significant differences in ctDNA detection rate. Furthermore, we found that DNA purified from BAL supernatants contains the highest levels of ctDNA compared with BAL precipitates and plasma; therefore, utilizing BAL supernatants for tumor detection might provide additional benefits. Finally, we demonstrated that tumor cellularity played significant roles in the design of personalized ctDNA panel which eventually impacts ctDNA detection sensitivity. We suggest setting a flexible criteria for sample quality control and utilization of BAL might benefit more patients in clinics.

Abstract Background The absence of thyroid transcription factor 1 (TTF‐1) is associated with a lower frequency of epidermal growth factor receptor (EGFR) mutations in lung adenocarcinoma (LUAD). The aim of this study was to assess the impact of TTF‐1 expression on the clinical response to EGFR‐tyrosine kinase inhibitor (TKI) treatment in patients with advanced LUAD. Methods The data of patients with advanced LUAD who were admitted to the Beijing Tiantan Hospital and Peking University Cancer Hospital (China) between April 2009 and May 2023 was retrospectively analyzed. Results A total of 227 patients diagnosed with advanced LUAD were included, of which 28.2% (64/227) had TTF‐1‐negative adenocarcinoma, while 54.6% (124/227) harbored EGFR mutations. Negative TTF‐1 expression significantly correlated with male sex (68.8% vs. 42.3%, p

Abstract Background Ubiquitin‐conjugating enzyme E2S (UBE2S), an E2 enzyme, is associated with the development of various tumors and exerts oncogenic activities. UBE2S is overexpressed in tumors, including hepatocellular carcinoma (HCC). However, the key molecular mechanisms of UBE2S in HCC still need additional research. The aim of this study was to explore the role of UBE2S in HCC. Methods The expression levels of UBE2S in HCC tissues and cells were detected by western blot analysis, quantitative real‐time polymerase chain reaction analysis (qRT–PCR), and immunohistochemistry (IHC). A 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide (MTT) assay, wound healing assay, colony formation assay transwell assay, and animal models were used to detect the proliferation and migration ability of HCC cells. Western blot analysis, qRT–PCR, immunofluorescence, small‐interfering RNA (siRNA), and plasmid transfection and coimmunoprecipitation (Co‐IP) assays were performed to detect the interaction among UBE2S, von Hippel–Lindau (VHL), hypoxia‐inducible factor 1‐alpha (HIF‐1α), Janus kinase‐2 (JAK2), and signal transducer and activator of transcription 3 (STAT3). Results In this study, we found that high UBE2S expression was associated with poor prognosis in HCC patients. In addition, UBE2S expression was upregulated in HCC tissues and cell lines. Knockdown of UBE2S inhibited the proliferation and migration of HCC cells in vitro and in vivo by directly interacting with VHL to downregulate the HIF‐1α and JAK2/STAT3 signaling pathways. Accordingly, overexpression of UBE2S significantly enhanced the proliferation and migration of HCC cells in vitro via VHL to upregulate HIF‐1α and JAK2/STAT3 signaling pathways. Furthermore, we found that downregulation of UBE2S expression enhanced the sensitivity of HCC cells to sorafenib in vivo and in vitro. Conclusion UBE2S enhances malignant properties via the VHL/HIF‐1α and VHL/JAK2/STAT3 signaling pathways and reduces sensitivity to sorafenib in HCC. The findings of this study may open a new approach for HCC diagnosis and provide a potential option for the treatment of HCC.

Summary: Lactic acid has emerged as an important modulator of immune cell function. It can be produced by both gut microbiota and the host metabolism at homeostasis and during disease states. The production of lactic acid in the gut microenvironment is vital for tissue homeostasis. In the present study, we examined how lactic acid integrates cellular metabolism to shape the epigenome of macrophages during pro-inflammatory response. We found that lactic acid serves as a primary fuel source to promote histone H3K27 acetylation, which allows the expression of immunosuppressive gene program including Nr4a1. Consequently, macrophage pro-inflammatory function was transcriptionally repressed. Furthermore, the histone acetylation induced by lactic acid promotes a form of long-term immunosuppression (“trained immunosuppression”). Pre-exposure to lactic acid induces lipopolysaccharide tolerance. These findings thus indicate that lactic acid sensing and its effect on chromatin remodeling in macrophages represent a key homeostatic mechanism that can provide a tolerogenic tissue microenvironment.

The structural, dynamical, and thermodynamic properties of zinc-blende (ZB) mercury selenide (HgSe) and cadmium selenide (CdSe) were systematically explored using an ab initio pseudopotential approach based on the density functional theory (DFT). The optimized lattice constants are in excellent agreement with the available experimental and theoretical results. The electronic properties show semiconducting behavior with a direct bandgap of 0 and 0.36 eV for HgSe and CdSe, respectively. Also, using the linear response approach, the detailed vibration properties of HgSe and CdSe are obtained. According to the calculated phonon dispersion, it is concluded that HgSe and CdSe are dynamically stable in the ground state; several peculiarities in the dispersion curves of these materials are generally similar, but the zone-boundary frequencies and zone-center optical frequencies of HgSe are lower than those of CdSe. The phonon frequencies at the high-symmetry points Γ, X, and L for HgSe and CdSe are in good agreement with the other theoretical reported. On the other hand, the dielectric properties are investigated and discussed. The results show that the dielectric tensor is diagonal, and the Born effective charge tensor is also isotropic. Finally, based on the lattice dynamical properties, we investigate the thermodynamic properties of both compounds, such as Helmholtz free energy F, internal energy E, entropy S, and specific heat Cv. The results show that HgSe and CdSe exhibit similar but slightly different behaviors in terms of its thermodynamic properties, which is expected because the atoms Hg and Cd are in the same group and exhibit a different atom mass. Most of the results presented in this study are novel and should provide a useful reference for future studies.

The workforce is aging with the population aging. How to effectively manage and motivate older workers is significant for elderly human resources development and the sustainable development of enterprises in organizations. Age stereotypes of older workers refer to people’s beliefs and expectations about a specific group of 45–65-year-olds in the workplace. This paper examines the effect of age stereotypes of older workers on job performance and intergenerational knowledge transfer intention. This study carried out two research designs, a questionnaire survey and an experimental study, to explore the effects of positive and negative age stereotypes of older workers on job performance and intergenerational knowledge transfer intention within an organizational context and underlying mediating mechanisms. The results showed that positive stereotypes of older workers significantly positively affected job performance and intergenerational knowledge transfer intention. In comparison, negative stereotypes of older workers significantly negatively affected job performance and intergenerational knowledge transfer intention, and self-perception of aging substantially mediates the effects. This study broadens the research field on the impact of positive and negative age stereotypes on older workers in organizational contexts. This study guides organizations in reducing age discrimination, creating an inclusive workplace environment, and achieving the successful aging of older workers.

Cholesterol levels were strongly associated with tumor progression and metastasis. Targeted cholesterol metabolism has broad prospects in tumor treatment. Ezetimibe, the only FDA-approved inhibitor of cholesterol absorption, has been reported to be able to inhibit angiogenesis in liver cancer. However, the efficacy and specific mechanisms of Ezetimibe in the treatment of Triple-Negative Breast Cancer (TNBC)have not been reported. Our research shows Ezetimibe inhibits TNBC cell proliferation and blocks the cell cycle in the G1 phase. Mechanistically, Ezetimibe inhibits the activation of PDGFRβ/AKT pathway, thereby promoting cell cycle arrest and inhibiting cell proliferation. By overexpressing PDGFRβ in TNBC cells, we found that PDGFRβ significantly reduced the inhibitory effect of Ezetimibe on TNBC cell proliferation and the cell cycle. Similarly, SC79, an AKT agonist, can reduce the proliferation inhibitory and cycle-blocking effects of Ezetimibe on TNBC cells. Furthermore, the AKT inhibitor MK2206 enhanced the inhibitory effect of Ezetimibe on the cell cycle and proliferation ability of TNBC cells overexpressing PDGFRβ. In xenograft tumor models, we also found that Ezetimibe inhibited TNBC growth, an effect that can be blocked by overexpression of PDGFR or activation of AKT. In summary, we have demonstrated that EZ inhibits the PDGFR/AKT pathway, thereby halting TNBC cycle progression and tumor growth.

Abstract Background Combinatorial systemic chemotherapy is a powerful treatment paradigm against cancer, but it is fraught with problems due to the emergence of chemoresistance and additive systemic toxicity. In addition, coadministration of individual drugs suffers from uncontrollable pharmacokinetics and biodistribution, resulting in suboptimal combination synergy. Methods Toward the goal of addressing these unmet medical issues, we describe a unique strategy to integrate multiple structurally disparate drugs into a self-assembling nanococktail platform. Conjugation of a polyunsaturated fatty acid (e.g., linoleic acid) with two chemotherapies generated prodrug entities that were miscible with tunable drug ratios for aqueous self-assembly. In vitro and in vivo assays were performed to investigate the mechanism of combinatorial nanococktails in mitigating chemoresistance and the efficacy of nanotherapy. Results The coassembled nanoparticle cocktails were feasibly fabricated and further refined with an amphiphilic matrix to form a systemically injectable and PEGylated nanomedicine with minimal excipients. The drug ratio incorporated into the nanococktails was optimized and carefully examined in lung cancer cells to maximize therapeutic synergy. Mechanistically, subjugated resistance by nanococktail therapy was achieved through the altered cellular uptake pathway and compromised DNA repair via the ATM/Chk2/p53 cascade. In mice harboring cisplatin-resistant lung tumor xenografts, administration of the nanococktail outperformed free drug combinations in terms of antitumor efficacy and drug tolerability. Conclusion Overall, our study provides a facile and cost-effective approach for the generation of cytotoxic nanoparticles to synergistically treat chemoresistant cancers.

Background: Ferroptosis is a novel process of programmed cell death driven by excessive lipid peroxidation that is associated with the development of lung adenocarcinoma. N6-methyladenosine (m6a) modification of multiple genes is involved in regulating the ferroptosis process, while the predictive value of N6-methyladenosine- and ferroptosis-associated lncRNA (FMRlncRNA) in the prognosis of patients remains with LUAD remains unknown.Methods: Unsupervised cluster algorithm was applied to generate subcluster in LUAD according to ferroptosis-associated lncRNA. Stepwise Cox analysis and LASSO algorithm were applied to develop a prognostic model. Cellular location was detected by single-cell analysis. Also, we conducted Gene set enrichment analysis (GSEA) enrichment, immune microenvironment and drug sensitivity analysis. In addition, the expression and function of the LINC01572 were investigated by several in vitro experiments including qRT-PCR, cell viability assays and ferroptosis assays.Results: A novel ferroptosis-associated lncRNAs-based molecular subtype containing two subclusters were determined in LUAD. Then, we successfully created a risk model according to five ferroptosis-associated lncRNAs (LINC00472, MBNL1-AS1, LINC01572, ZFPM2-AS1, and TMPO-AS1). Our nominated model had good stability and predictive function. The expression patterns of five ferroptosis-associated lncRNAs were confirmed by polymerase chain reaction (PCR) in LUAD cell lines. Knockdown of LINC01572 significantly inhibited cell viability and induced ferroptosis in LUAD cell lines.Conclusion: Our data provided a risk score system based on ferroptosis-associated lncRNAs with prognostic value in LUAD. Moreover, LINC01572 may serve as a novel ferroptosis suppressor in LUAD.

A bit-level object storage system is a foundational building block of long-term digital preservation (LTDP). To achieve the purposes of LTDP, the system must be able to: preserve the authenticity and integrity of the original digital objects; scale up with dramatically increasing demands for preservation storage; mitigate the impact of hardware obsolescence and software ephemerality; replicate digital objects among distributed data centers at different geographical locations; and to constantly audit and automatically recover from compromised states. A realistic and daunting challenge to satisfy these requirements is not only to overcome technological difficulties but also to maintain economic sustainability by implementing and continuously operating such systems in a cost-effective way. In this paper, we present OpenStack Swift, an open-source, mature and widely accepted cloud platform, as a practical and proven solution with a case study at the University of Alberta Library. We emphasize the implementation, application, cost analysis and maintenance of the system, with the purpose of contributing to the community with an exceedingly robust, highly scalable, self-healing and comparatively cost-effective bit-level object storage system for long-term digital preservation.

Peroxidase (PXDN), a specific extracellular matrix (ECM)-associated protein, has been determined as a tumor indicator and therapeutic target in various tumors. However, the effects of PXDN in prognostic performance and clinical implications in glioblastoma multiforme (GBM) remains unknown. Here, we assessed PXDN expression pattern and its performance on prognosis among GBM cases from TCGA and CGGA databases. PXDN was up-regulated within GBM samples in comparison with normal control. High PXDN expression was a dismal prognostic indicator in GBM. Single cell RNA analysis was conducted to detect the cell localization of PXDN. We also set up a PPI network to explore the interacting protein associated with PXDN, including TSKU, COL4A1 and COL5A1. Consistently, functional enrichment analysis revealed that several cancer hallmarks were enriched in the GBM cases with high PXDN expression, such as epithelial-mesenchymal transition (EMT), fatty acid metabolism, glycolysis, hypoxia, inflammatory response, and Wnt/beta-catenin signaling pathway. Next, this study analyzed the association of PXDN expression and immunocyte infiltration. PXDN expression was in direct proportion to the infiltrating degrees of NK cells resting, T cells regulatory, M0 macrophage, monocytes and eosinophils. The roles of PXDN on immunity were further estimated by PXDN-associated immunomodulators. In addition, four prognosis-related lncRNAs co-expressed with PXDN were identified. Finally, we observed that PXDN depletion inhibits GBM cell proliferation and migration by in vitro experiments. Our data suggested that PXDN has the potential to be a powerful prognostic biomarker, which might offer a basis for developing therapeutic targets for GBM.

Abstract We focus on the causes of fluctuations in wintertime PM10 in nine regional core cities of China using two machine learning models, Random Forest (RF) and Recurrent Neural Network (RNN). RF and RNN both show high performance in predicting hourly PM10 using only gaseous air pollutants (SO2, NO2 and CO) as inputs, showing the predominance of the secondary inorganic aerosol and implying the existence of thermodynamic equilibrium between gaseous air pollutants and PM10. Also, we find the following results. The correlation of gaseous air pollutants and PM10 were more relevant than that of meteorological conditions and PM10. CO was the predominant factor for PM10 in the Beijing-Tianjin-Hebei Plain and the Yangtze River Delta while SO2 and NO2 were also important features for PM10 in the Pearl River Delta and Sichuan Basin. The spatial heterogeneity and temporal homogeneity of PM10 in China are revealed. The long-range transported PM10 was substantiated to be insignificant, except in the sandstorms. The severity of PM10 was attributable to the lopsided shift of thermodynamic equilibrium and the phenology of indigenous flora.

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In this paper we deal with a system of partial differential equations describing a steady motion of an incompressible magnetohydrodynamic fluid, where the extra stress tensor is induced by a potential with $p$-structure ($p=2$ corresponds to the Newtonian case). By using a fixed point argument in an appropriate functional setting, we proved the existence and uniqueness of strong solutions for the problem in a smooth domain $\Omega\subset\mathbb{R}^{n}$ $(n=2,3)$ under the conditions that the external force is small in a suitable norm.

Since the reform and opening up, China’s rapid economic growth mainly depends on the industrial development mode of “high energy consumption and high pollution,” which has caused serious haze pollution. In order to achieve the goal of haze control and sustainable development, we need to give full play to the role of technological innovation. Empirical analysis of the haze control effect of technological innovation has theoretical significance and practical value. Based on the panel data of 30 provinces in China from 2005 to 2018 and the PM2.5 concentration data published by the atmospheric composition analysis group of Dalhousie University, this study selects R&D personnel input and technology market turnover to represent the level of technological innovation and uses the panel data model, threshold effect model, and spatial Durbin model to empirically analyze the impact of technological innovation on haze pollution control. The empirical results show that 1) technological innovation can significantly reduce the PM2.5 concentration of the province, showing a positive haze control effect; 2) technological innovation indicates a negative indirect effect on PM2.5 concentration, confirming the “technology spillover effect,” that is, technological innovation also has a haze control effect on the surrounding provinces; 3) with the increase in the province’s economic aggregate, the haze control effect of technological innovation shows a trend of “high low high,” and the role of technological innovation is the lowest in the stage of economic transformation; and 4) from the perspective of regional differentiation, the haze control effect of technological innovation is the largest in the central region, and the smallest in the western region. Technological innovation indicates a positive haze control effect on all regions at all stages of economic development. This study provides policy suggestions for the government and enterprises to use innovation for cleaner production and sustainable development.

The development and utilization of 3p-block based MOFs as fluorescent materials has attracted significant attention in recent years. Herein, we have successfully constructed a versatile luminescent Ga-MOF (SNNU-63) with a 3d10 configuration and a large ligand twist configuration. Interestingly, the as-synthesized Ga-MOF exhibits excellent luminescence property and a good material for blue light-emitting diode (LED). At 80 K, this Ga-MOF shows multi-emission centers at 381, 462, and 494 nm. As a ratiometric thermometer, this Ga-MOF exhibits an excellent temperature sensing property with high relative sensitivity (Sm = 2.60 % K−1 at 110 K). The fluorescence intensity ratio I381/I494 shows a very good fit for the Boltzmann results (80–240 K). Moreover, the luminescent Ga-MOF exhibits an excellent selective detection of Fe3+ over other metal ions in aqueous an medium, and the limit of detection (LOD) towards Fe3+ ions is calculated to be 1.227 × 10−4 M. This work presents a versatile luminescent Ga-MOF material as a blue LED and fluorescent probe for low-temperature and selective Fe3+ sensing.

Abstract Background Immune‐therapy with anti‐PD1 inhibitors, such as pembrolizumab, is revolutionizing the treatment of non‐small cell lung cancers (NSCLC). However, identifying patients for the potential therapeutic response and predicting therapy resistance and early relapse remains a challenge. Methods Between 2016 and 2018, 60 patients were treated with pembrolizumab, among who 12 NSCLC patients had both baseline (before treatment) and serial (on treatment) periodical circulating tumor DNA (ctDNA) samples. Those samples were sequenced on a 329 pan cancer‐related gene panel. Analyses of tumor burden, blood tumor mutational burden (bTMB), maximum somatic allele frequency (MSAF), and tumor clonal structure were performed in association with clinical response. Candidate resistance mutations involved in relapse and metastases were further investigated. Results ctDNA was detected and mutational profiling was performed for each patient. Those with a high baseline bTMB level showed significantly improved progression‐free survival (PFS) after pembrolizumab treatment. Tumor burden and therapeutic response significantly correlated with the MSAF instead of the bTMB. Clone analysis detected tumor progression about 2‐4 months ahead of computed tomography (CT) scan. One mutation in gene PTCH1 (Protein patched homolog 1) and two acquired anti‐PD1 candidate resistance mutations of gene B2M (β2 microglobulin) were identified in association with distant metastasis. The evolutionary tree of a representative patient was also described. Conclusion This pilot study showed that MSAF could be another good indicator of therapeutic response, and clonal analysis could be clinically useful in monitoring clonal dynamics and detecting remote metastasis and early relapse.

Abstract Background Lung adenocarcinoma in young adults is a rare entity with the oncogenic genetic alterations associated being poorly understood. In the present study, the effect of genetic alterations in lung adenocarcinoma patients diagnosed in young patients is reported. Methods Twenty young lung adenocarcinoma patients (age years: median: 33.5, range: 24‐36) were enrolled in the current study and 24 patients who were at common age of the disease onset (age years: median: 61.5, range: 52‐79) were selected for comparison. Paraffin sections of lung adenocarcinoma were analyzed using the whole‐exome sequencing platform. Results Similar number of somatic mutations per tumor were found in the young patients and their older counterparts. Although no age‐related differences were detected in the numbers of lung adenocarcinoma patients harboring well‐known gene variants, mutations in FRG1 and KMT2C were associated with a younger age especially after correcting for tobacco smoking and sex (FRG1: P = 0.027, KMT2C: P = 0.046). Five genetic variants showed higher alteration frequencies in young patients compared to the unclassified East Asian population, suggesting these mutations as disease‐related hereditary germline variants. Conclusions These results suggest different characteristics of lung adenocarcinoma between the young and the patients at common age of onset. Young patients with lung adenocarcinoma have a distinctly unique prevalence of oncogenic genetic alterations.

In wireless sensor networks (WSNs), energy efficiency and low delay are two pivotal issues for data collection. Wireless sensor networks are composed of energy-constrained sensor nodes. While sensing the environment, the rapid transmission of data to the sink is critical for many applications. In order to save energy, the duty cycle based mode is adopted in most studies, but this method increases the data transmission delay. Thus, an Adding Duty Cycle Only in Connected Dominating Sets (ADCOCDS) approach is proposed in this paper, which can effectively reduce network energy consumption and delay. The main innovations of the ADCOCDS scheme are as follows: (1) Unlike the mode of the duty cycle for all nodes in the previous WSNs, in ADCOCDS scheme, we choose a part of nodes to construct as a connected dominating set, and these nodes adopt the mode of duty cycle. For other nodes in the network, the radio transmitter is turned off to save energy when there is no data to transmit, and when these nodes have data to send, they turn on the radio transmitter and send the data to nodes in the dominating set, then nodes in the dominating set transfer the data to sink. In ADCOCDS scheme, since only a small number of nodes adopt the duty cycle mode, the energy efficiency of the network is high; (2) In WSNs, we observed that the energy consumption of nodes in the near-sink area is higher than that in the far-sink area. Therefore, in the ADCOCDS scheme, we increase the duty cycle of nodes according to their energy consumption to reduce the delay. Through theoretical analysis and experimental results, the ADCOCDS scheme can effectively reduce energy consumption and delay by 12.09%-43.17% and 21.85%-57.96%, respectively, while improving network lifetime by 10.71%-24.13% compared with previous strategies.

Background: Data on inter-tumoral heterogeneity and clonal evolution of pancreatic neuroendocrine neoplasms (panNENs) with liver metastasis are limited. The aim of this study was to explore different patterns of clonal evolution of pancreatic neuroendocrine neoplasms with liver metastasis and the possible distinctive signaling pathways involved between G2 neuroendocrine tumors (NETs) and neuroendocrine carcinomas (NECs).Methods: Tumor tissues of five patients (10 samples) with pancreatic neuroendocrine neoplasms with synchronous liver metastasis were analyzed using next-generation sequencing. PyClone, Gene Ontology, and Reactome pathway enrichment analysis were also applied.Results: Mutated genes varied in individuals, reflecting the inter-tumoral heterogeneity of panNENs. The distribution of subclones varied during tumor metastasis, and different clonal evolution patterns were revealed between NETs and NECs. Gene Ontology and Reactome analyses revealed that in both NETs and NECs, signaling pathways and biological processes shared similarities and differences in the primary and metastatic lesions. In addition, the signaling pathway features were different between NETs and NECs. In the primary lesions, epigenetic changes and post-transcriptional modifications participated in NETs, while FGFR signaling, EGFR signaling, and NTRK2 signaling were largely involved in NECs. Although DNA repair and TP53 regulation were both involved in the metastatic lesions, most of the signaling pathways and biological processes disrupted by the mutated genes were different.Conclusions: Our study revealed spatial inter-tumoral heterogeneity and temporal clonal evolution in PanNENs, providing potential therapeutic targets for further prospective clinical trials.

BackgroundTriple-negative breast cancer (TNBC) features a poor prognosis, which is partially attributed to its high metastatic rate. However, there is no effective target for systemic TNBC therapy due to the absence of estrogen, progesterone, and human epidermal growth factor 2 receptors (ER, PR, and HER-2, respectively) in cancer. In the present study, we evaluated the role of sphingosine kinase 2 (SphK2) and its catalyst sphingosine-1-phosphate (S1P) in TNBC metastasis and the effect of the SphK2-specific inhibitor ABC294640 on TNBC metastasis.MethodsThe function of SphK2 and S1P in TNBC cell metastasis was evaluated using transwell migration and wound-healing assays. The molecular mechanism of SphK2/S1P mediating TNBC metastasis was investigated using Western blot, histological examination, and immunohistochemistry assays. The antitumor activity of ABC294640 was examined in an in vivo TNBC lung metastatic model.ResultsSphingosine kinase 2 promoted TNBC cell migration through the generation of S1P. Targeting SphK2 with ABC294640 inhibited TNBC lung metastasis in vivo. p21-activated kinase 1 (PAK1), p-Lin-11/Isl-1/Mec-3 kinase 1 (LIMK1), and Cofilin1 were the downstream signaling molecules of SphK2/S1P. Inhibition of PAK1 suppressed SphK2/S1P-induced TNBC cell migration.ConclusionSphingosine kinase 2/sphingosine-1-phosphate promotes TNBC metastasis through the activation of the PAK1/LIMK1/Cofilin1 signaling pathway. ABC294640 inhibits TNBC metastasis in vivo and could be developed as a novel agent for the clinical treatment of TNBC.

Al-Si-Mg-Cu-Ni alloy is widely used in the manufacture of high-performance car engine parts. Coarse, dendritic α-Al and large primary Si are common in Al-Si-Mg-Cu-Ni alloy DC casting billet, which is harmful to the performance of the final product. In this paper, a pulsed magnetic field melt treatment technique was applied to the melt in the launder of a DC casting platform to modify the α-Al and primary Si in the billet. A transient numerical model was established to analyze the electromagnetic field, flow field and temperature field in the melt during the pulsed magnetic field treatment. The effect of the magnetic energy on the clusters in the melt was analyzed. We found that during the pulsed magnetic field melt treatment, the number of clusters close to the critical size was increased due to the cluster formation work being reduced by the magnetic energy, which facilitated nucleation and refined the solidification structure. Furthermore, the flow velocity increased, and temperature homogenized in the melt during the pulsed magnetic field melt treatment, which benefitted the clusters close to the critical size distributed and maintained in the melt uniformly. The experimental results show that the α-Al and primary Si were small and homogeneous following the pulsed magnetic field melt treatment. The size of α-Al and primary Si was reduced by 25.6–44.4% and 32.2–54.1%, respectively, in the billet center compared to the conventional process.

Background: Regorafenib is a second-line therapy drug used for advanced hepatocellular carcinoma (HCC). Unfortunately, the survival benefit of the patients receiving this treatment is modest, which may be attributed to drug resistance. In the present study, sphingosine kinase 2 (SphK2) was targeted to reverse regorafenib resistance in HCC.Methods: The functions of SphK2 and sphingosine-1-phosphate (S1P), the catalytic product of SphK2 in regorafenib resistance of HCC cells, were evaluated by cell counting kit-8 assay, colony formation, cell cycle evaluation, and annexin V–fluorescein isothiocyanate/propidium iodide double-staining assay. The antitumor activity of combined treatment of regorafenib and the SphK2-specific inhibitor ABC294640 was examined in HCC cells in vitro and xenograft model in vivo. The molecular mechanisms of SphK2/S1P-mediating regorafenib resistance were investigated using cell line establishment and Western blot analysis.Results: Well-developed regorafenib-resistant HCC cells indicated high expression levels of SphK2. The sensitivity to regorafenib of regorafenib-resistant HCC cells was restored following SphK2 knockdown or pharmacological inhibition by ABC294640. In addition, ectopic expression of SphK2 and exogenous addition of S1P decreased the sensitivity of HCC cells to regorafenib. Furthermore, the combination treatment with ABC294640 sensitized resistant tumor to regorafenib in xenograft model of HCC. The phosphorylation levels of nuclear factor κB (NF-κB), as well as those of signal transducer and activator of transcription 3 (STAT3), were positively associated with SphK2 and S1P.Conclusions: SphK2/S1P mediates regorafenib resistance of HCC through NF-κB and STAT3 activation. Targeting SphK2 by ABC294640 potently reduces regorafenib resistance of HCC cells both in vitro and in vivo. The combination of ABC294640 and regorafenib could be developed as a novel potential treatment strategy for advanced HCC.

Silicon carbide (SiC) ceramics, as a superhard nonmetallic material, would induce severe tool wear during drilling. To ensure the quality of processing, the drill bits must be replaced in advance before failure, which leads to serious waste without effective usage. Aiming at the tool reliability problem determined by the tool wear during drilling of the high hardness nonmetallic materials, this article proposed a reliability analysis method of sintered diamond drill bit for machining SiC ceramics based on the distribution of tool degradation increment and the modeling of gamma process, and a tool reliability model of degradation increment was established based on the Bayesian theory. The drilling experiment of sintered SiC ceramics was carried out by sintering diamond drill bit to obtain the degradation increment data. The accuracy of the model was tested by the reliability model deviation rate, which indicated that the reliability model with considering individual differences was more accurate. This research can provide a theoretical model and analytical method for improving the reliability of the nonmetallic brittle material processing tools.

Summary: Multi-region sequencing is used to detect intratumor genetic heterogeneity (ITGH) in tumors. To assess whether genuine ITGH can be distinguished from sequencing artifacts, we performed whole-exome sequencing (WES) on three anatomically distinct regions of the same tumor with technical replicates to estimate technical noise. Somatic variants were detected with three different WES pipelines and subsequently validated by high-depth amplicon sequencing. The cancer-only pipeline was unreliable, with about 69% of the identified somatic variants being false positive. Even with matched normal DNA for which 82% of the somatic variants were detected reliably, only 36%–78% were found consistently in technical replicate pairs. Overall, 34%–80% of the discordant somatic variants, which could be interpreted as ITGH, were found to constitute technical noise. Excluding mutations affecting low-mappability regions or occurring in certain mutational contexts was found to reduce artifacts, yet detection of subclonal mutations by WES in the absence of orthogonal validation remains unreliable. : Shi et al. report that standard coverage whole-exome sequencing and bioinformatics pipelines cannot discriminate between genuine intratumor genetic heterogeneity and sequencing artifacts. Although aggressive minimum depth filtering would not improve the false detection rate of subclonal mutations, excluding mutations in low-mappability regions or in certain mutational contexts could help. Keywords: massively parallel sequencing, whole-exome sequencing, somatic mutations, intratumor genetic heterogeneity, multi-region profiling, breast cancer, mutational signatures, mappability, subclonal

Closed-loop stimulation of many neurological disorders, such as epilepsy, is an emerging technology and regarded as a promising alternative for surgical and drug treatment. In this paper, a real-time seizure detection algorithm based on STFT and support vector machine (SVM) and its field-programmable gate array (FPGA) implementation are proposed. With a two-stage patient-specific channel selection and feature selection mechanism, those redundant and uncorrelated spectral features are removed from the entire feature set. The evaluation results on CHB-MIT epilepsy database show that the mean detection latency of the proposed algorithm is 6 s, the sensitivity is 98.4%, and the false detection rate is 0.356/h. The performance of our proposed algorithm is comparable to other existing seizure detection algorithms. Moreover, we implement the proposed seizure detection algorithm on Xilinx Zynq-7000 XC7Z020 with high level synthesis. Each classification of the input electroencephalography signal can be finished within 313 $\mu \text{s}$ , and the power consumption of the programmable logic is only 380 mW at 100 MHz. In hardware implementation, an optimization strategy for the nested-loop structure within nonlinear SVM is proposed to improve pipeline efficiency. Compared with existing method, the experimental result shows that our method can speed up the nonlinear SVM by $1.70\times $ , $1.53\times $ , $1.37\times $ , and $1.26\times $ with the unroll factor equal to 1–4 at the same DSP utilization rate. The evaluation results affirm the possibility of integrating the proposed algorithm and FPGA implementation into a wearable seizure control device.

Hexavalent chromium contamination in groundwater has become a very serious and challenging problem. Identification of the groundwater chemical characteristics of the sites and their control mechanisms for remediation of pollutants is a significant challenge. In this study, a contaminated site in Xinxiang City, Henan Province, was investigated and 92 groundwater samples were collected from the site. Furthermore, the hydrogeochemical characteristics and the distribution patterns of components in the groundwater were analyzed by a combination of multivariate statistical analysis, Piper diagram, Gibbs diagram, ions ratio and hydrogeochemical simulation. The results showed that the HCO3-Cl-Mg-Ca type, SO4-HCO3-Na type, and HCO3-Mg-Ca-Na type characterize the hydrogeochemical composition of shallow groundwater and HCO3-Cl-Mg-Ca type, HCO3-Na-Mg type, and HCO3-SO4-Mg-Na-Ca type characterize the hydrogeochemical composition of deep groundwater. Ion ratios and saturation index indicated that the groundwater hydrogeochemical characteristics of the study area are mainly controlled by water–rock action and evaporative crystallization. The dissolution of halite, gypsum and anhydrite, the precipitation of aragonite, calcite and dolomite, and the precipitation of trivalent chromium minerals other than CrCl3 and the dissolution of hexavalent chromium minerals occurred in groundwater at the site. The minimum value of pH in groundwater at the site is 7.55 and the maximum value is 9.26. The influence of pH on the fugacity state of minerals was further investigated. It was concluded that the saturation index of dolomite, calcite, aragonite and MgCr2O4 increases with the increase of pH, indicating that these minerals are more prone to precipitation, and the saturation index of Na2Cr2O7, K2Cr2O7 and CrCl3 decreases with the increase of pH, implying that Na2Cr2O7, K2Cr2O7 and CrCl3 are more prone to dissolution. The saturation index of the remaining minerals is less affected by pH changes. The study can provide a scientific basis for groundwater remediation.

Data missing in collections of time series occurs frequently in practical applications and turns out to be a major menace to precise data analysis. However, most of the existing methods either might be infeasible or could be inefficient to predict the missing values in large-scale coevolving time series. Also, the evolving of time series needs to be handled properly to adapt to the temporal characteristic. Furthermore, more massive volume of data is generated in many areas than ever before. In this paper, we have taken up the challenge of missing data prediction in coevolving time series by employing temporal dynamic matrix factorization techniques. First, our approaches are optimally designed to largely utilize both the interior patterns of each time series and the information of time series across multiple sources to build an initial model. Based on the idea, we have imposed hybrid regularization terms to constrain the objective functions of matrix factorization. Then, temporal dynamic matrix factorization is proposed to effectively update the initial already trained models. In the process of dynamic matrix factorization, batch updating and fine-tuning strategies are also employed to build an effective and efficient model. Extensive experiments on real-world data sets and synthetic data set demonstrate that the proposed approaches can effectively improve the performance of missing data prediction. Even when the missing ratio reaches as high as 90%, our proposed methods still show low prediction errors. Dynamic performance demonstrates that the methods can obtain satisfactory effectiveness and efficiency. Furthermore, we have also demonstrated how to take advantage of the high processing power of Apache Spark to perform missing data prediction in large-scale coevolving time series.

Background Undersized ring annuloplasty is a commonly used surgical repair for ischemic mitral regurgitation, in which annular downsizing corrects regurgitation, but alters valve geometry and elevates tissue stresses. In this study, we investigated if unphysiological leaflet kinematics after annuloplasty might cause pathogenic biological remodeling of the mitral valve leaflets, and if preserving physiologic leaflet kinematics with a better technique can moderate such adverse remodeling. Methods and Results Twenty‐nine swine were induced with ischemic mitral regurgitation, and survivors were assigned to groups: 7 underwent annuloplasty, 12 underwent annuloplasty with papillary‐muscle approximation, 6 underwent papillary‐muscle approximation, and 3 were sham controls. Pre‐and post‐surgery leaflet kinematics were measured, and valve tissue was explanted after 3 months to assess biological changes. Anterior leaflet excursion was unchanged across groups, but persistent tethering was observed with annuloplasty. Posterior leaflet was vertically immobile after annuloplasty, better mobile with the combined approach, and substantially (P=0.0028) mobile after papillary‐muscle approximation. Procollagen‐1 was higher in leaflets from annuloplasty compared with the other groups. Heat shock protein‐47 and lysyl oxidase were higher in groups receiving annuloplasty compared with sham. α‐SMA was elevated in leaflets from animals receiving an annuloplasty, indicating activation of quiescent valve interstitial cells, paralleled by elevated transforming growth factor‐β expression. Conclusions This is the first study to demonstrate that surgical valve repairs that impose unphysiological leaflet mechanics have a deleterious, pathological impact on valve biology. Surgeons may need to consider restoring physiologic leaflet stresses as well as valve competence, while also exploring pharmacological methods to inhibit the abnormal signaling cascades.

BackgroundTriple negative breast cancer (TNBC) is a highly heterogeneous and aggressive disease, and although no effective targeted therapies are available to date, about one-third of patients with TNBC achieve pathologic complete response (pCR) from standard-of-care anthracycline/taxane (ACT) chemotherapy. The heterogeneity of these tumors, however, has hindered the discovery of effective biomarkers to identify such patients.Methods and findingsWe performed whole exome sequencing on 29 TNBC cases from the MD Anderson Cancer Center (MDACC) selected because they had either pCR (n = 18) or extensive residual disease (n = 11) after neoadjuvant chemotherapy, with cases from The Cancer Genome Atlas (TCGA; n = 144) and METABRIC (n = 278) cohorts serving as validation cohorts. Our analysis revealed that mutations in the AR- and FOXA1-regulated networks, in which BRCA1 plays a key role, are associated with significantly higher sensitivity to ACT chemotherapy in the MDACC cohort (pCR rate of 94.1% compared to 16.6% in tumors without mutations in AR/FOXA1 pathway, adjusted p = 0.02) and significantly better survival outcome in the TCGA TNBC cohort (log-rank test, p = 0.05). Combined analysis of DNA sequencing, DNA methylation, and RNA sequencing identified tumors of a distinct BRCA-deficient (BRCA-D) TNBC subtype characterized by low levels of wild-type BRCA1/2 expression. Patients with functionally BRCA-D tumors had significantly better survival with standard-of-care chemotherapy than patients whose tumors were not BRCA-D (log-rank test, p = 0.021), and they had significantly higher mutation burden (p < 0.001) and presented clonal neoantigens that were associated with increased immune cell activity. A transcriptional signature of BRCA-D TNBC tumors was independently validated to be significantly associated with improved survival in the METABRIC dataset (log-rank test, p = 0.009). As a retrospective study, limitations include the small size and potential selection bias in the discovery cohort.ConclusionsThe comprehensive molecular analysis presented in this study directly links BRCA deficiency with increased clonal mutation burden and significantly enhanced chemosensitivity in TNBC and suggests that functional RNA-based BRCA deficiency needs to be further examined in TNBC.

Nucleotide alterations detected by next generation sequencing are not always true biological changes but could represent sequencing errors. Even highly accurate methods can yield substantial error rates when applied to millions of nucleotides. In this study, we examined the reproducibility of nucleotide variant calls in replicate sequencing experiments of the same genomic DNA. We performed targeted sequencing of all known human protein kinase genes (kinome) (~3.2 Mb) using the SOLiD v4 platform. Seventeen breast cancer samples were sequenced in duplicate (n=14) or triplicate (n=3) to assess concordance of all calls and single nucleotide variant (SNV) calls. The concordance rates over the entire sequenced region were >99.99%, while the concordance rates for SNVs were 54.3-75.5%. There was substantial variation in basic sequencing metrics from experiment to experiment. The type of nucleotide substitution and genomic location of the variant had little impact on concordance but concordance increased with coverage level, variant allele count (VAC), variant allele frequency (VAF), variant allele quality and p-value of SNV-call. The most important determinants of concordance were VAC and VAF. Even using the highest stringency of QC metrics the reproducibility of SNV calls was around 80% suggesting that erroneous variant calling can be as high as 20-40% in a single experiment. The sequence data have been deposited into the European Genome-phenome Archive (EGA) with accession number EGAS00001000826.

Micro RNAs (miRNAs) are important regulators involved in various physical and pathological processes, including cancer. The miRNA-302 family has been documented as playing a critical role in carcinogenesis. In this study, we investigated the role of miRNA-302a in colon cancer. MiRNA-302a expression was detected in 44 colon cancer tissues and 10 normal colon tissues, and their clinicopathological significance was analyzed. Cell proliferation and cell cycle analysis were performed on colon cancer cells that stably expressed miRNA-302a. The target gene of miRNA-302a and the downstream pathway were further investigated. Compared with normal colon tissues, miRNA-302a expression was downregulated in colon cancer tissues. Overexpression of miRNA-302a induced G1/S cell cycle arrest in colon cancer cells, and suppressed colon cancer cell proliferation both in vitro and in vivo. Furthermore, miRNA-302a inhibited AKT expression by directly binding to its 3' untranslated region, resulting in subsequent alterations of the AKT-GSK3β-cyclin D1 pathway. These results reveal miRNA-302a as a tumor suppressor in colon cancer, suggesting that miRNA-302a may be used as a potential target for therapeutic intervention in colon cancer.

The title compound, [Pb2(C7H5O2)2(NO3)2(C12H8N2)2(H2O)2], crystallizes as a dinuclear centrosymmetric dimer containing two PbII atoms bridged by two benzoate ligands. Each PbII atom is seven-coordinated by a water molecule, a nitrate anion, a 1,10-phenanthroline (phen) ligand and two benzoate anions. The crystal packing is stabilized by O—H...O hydrogen bonds and by π–π stacking between neighboring phen ligands, with a centroid–centroid distance of 3.557 (3) Å.

REST is abundantly expressed in mouse embryonic stem cells (ESCs). Many genome-wide analyses have found REST to be an integral part of the ESC pluripotency network. However, experimental systems have produced contradictory findings: (1) REST is required for the maintenance of ESC pluripotency and loss of REST causes increased expression of differentiation markers, (2) REST is not required for the maintenance of ESC pluripotency and loss of REST does not change expression of differentiation markers, and (3) REST is not required for the maintenance of ESC pluripotency but loss of REST causes decreased expression of differentiation markers. These reports highlight gaps in our knowledge of the ESC network.Employing biochemical and genome-wide analyses of various culture conditions and ESC lines, we have attempted to resolve some of the discrepancies in the literature.We show that Rest+/- and Rest-/- AB-1 mutant ESCs, which did not exhibit a role of REST in ESC pluripotency when cultured in the presence of feeder cells, did show impaired self-renewal when compared with the parental cells under feeder-free culture conditions, but only in early passage cells. In late passage cells, both Rest+/- and Rest-/- AB-1 ESCs restored pluripotency, suggesting a passage and culture condition-dependent response. Genome-wide analysis followed by biochemical validation supported this response and further indicated that the restoration of pluripotency was associated by increased expression of the ESC pluripotency factors. E14Tg2a.4 ESCs with REST-knockdown, which earlier showed a REST-dependent pluripotency when cultured under feeder-free conditions, as well as Rest-/- AB-1 ESCs, showed no REST-dependent pluripotency when cultured in the presence of either feeder cells or laminin, indicating that extracellular matrix components can rescue REST's role in ESC pluripotency.REST regulates ESC pluripotency in culture condition- and ESC line-dependent fashion and ESC pluripotency needs to be evaluated in a context dependent manner.