Search

Abstract Background In the field of ornamental horticulture, phenotypic mutations, particularly in leaf color, are of great interest due to their potential in developing new plant varieties. The introduction of variegated leaf traits in plants like Heliopsis helianthoides, a perennial herbaceous species with ecological adaptability, provides a rich resource for molecular breeding and research on pigment metabolism and photosynthesis. We aimed to explore the mechanism of leaf variegation of Heliopsis helianthoides (using HY2021F1-0915 variegated mutant named HY, and green-leaf control check named CK in 2020 April, May and June) by analyzing the transcriptome and metabolome. Results Leaf color and physiological parameters were found to be significantly different between HY and CK types. Chlorophyll content of HY was lower than that of CK samples. Combined with the result of Weighted Gene Co-expression Network Analysis (WGCNA), 26 consistently downregulated differentially expressed genes (DEGs) were screened in HY compared to CK subtypes. Among the DEGs, 9 genes were verified to be downregulated in HY than CK by qRT-PCR. The reduction of chlorophyll content in HY might be due to the downregulation of FSD2. Low expression level of PFE2, annotated as ferritin-4, might also contribute to the interveinal chlorosis of HY. Based on metabolome data, differential metabolites (DEMs) between HY and CK samples were significantly enriched on ABC transporters in three months. By integrating DEGs and DEMs, they were enriched on carotenoids pathway. Downregulation of four carotenoid pigments might be one of the reasons for HY’s light color. Conclusion FSD2 and PFE2 (ferritin-4) were identified as key genes which likely contribute to the reduced chlorophyll content and interveinal chlorosis observed in HY. The differential metabolites were significantly enriched in ABC transporters. Carotenoid biosynthesis pathway was highlighted with decreased pigments in HY individuals. These findings not only enhance our understanding of leaf variegation mechanisms but also offer valuable insights for future plant breeding strategies aimed at preserving and enhancing variegated-leaf traits in ornamental plants.

Abstract Background Dilated cardiomyopathy (DCM) is one of the most common causes of heart failure. Multiple identified mutations in nexilin (NEXN) have been suggested to be linked with severe DCM. However, the exact association between multiple mutations of Nexn and DCM remains unclear. Moreover, it is critical for the development of precise and effective therapeutics in treatments of DCM. Results In our study, Nexn global knockout mice and mice carrying human equivalent G645del mutation are studied using functional gene rescue assays. AAV-mediated gene delivery is conducted through systemic intravenous injections at the neonatal stage. Heart tissues are analyzed by immunoblots, and functions are assessed by echocardiography. Here, we identify functional components of Nexilin and demonstrate that exogenous introduction could rescue the cardiac function and extend the lifespan of Nexn knockout mouse models. Similar therapeutic effects are also obtained in G645del mice, providing a promising intervention for future clinical therapeutics. Conclusions In summary, we demonstrated that a single injection of AAV-Nexn was capable to restore the functions of cardiomyocytes and extended the lifespan of Nexn knockout and G645del mice. Our study represented a long-term gene replacement therapy for DCM that potentially covers all forms of loss-of-function mutations in NEXN.

Targeting multiple immune mechanisms may overcome therapy resistance and further improve cancer immunotherapy for humans. Here, we describe the application of virus-like vesicles (VLV) for delivery of three immunomodulators alone and in combination, as a promising approach for cancer immunotherapy. VLV vectors were designed to deliver single chain interleukin (IL)-12, short-hairpin RNA (shRNA) targeting programmed death ligand 1 (PD-L1), and a dominant-negative form of IL-17 receptor A (dn-IL17RA) as a single payload or as a combination payload. Intralesional delivery of the VLV vector expressing IL-12 alone, as well as the trivalent vector (designated CARG-2020) eradicated large established tumors. However, only CARG-2020 prevented tumor recurrence and provided long-term survival benefit to the tumor-bearing mice, indicating a benefit of the combined immunomodulation. The abscopal effects of CARG-2020 on the non-injected contralateral tumors, as well as protection from the tumor cell re-challenge, suggest immune-mediated mechanism of protection and establishment of immunological memory. Mechanistically, CARG-2020 potently activates Th1 immune mechanisms and inhibits expression of genes related to T cell exhaustion and cancer-promoting inflammation. The ability of CARG-2020 to prevent tumor recurrence and to provide survival benefit makes it a promising candidate for its development for human cancer immunotherapy.

BackgroundInterleukin-17 (IL-17) family cytokines promote protective inflammation for pathogen resistance, but also facilitate autoimmunity and tumor development. A direct signal of IL-17 to regulatory T cells (Tregs) has not been reported and may help explain these dichotomous responses.MethodsWe generated a conditional knockout of Il17ra in Tregs by crossing Foxp3-YFP-Cre mice to Il17ra-flox mice (Il17raΔTreg mice). Subsequently, we adoptively transferred bone marrow cells from Il17raΔTreg mice to a mouse model of sporadic colorectal cancer (Cdx2-Cre+/ApcF/+), to selectively ablate IL-17 direct signaling on Tregs in colorectal cancer. Single cell RNA sequencing and bulk RNA sequencing were performed on purified Tregs from mouse colorectal tumors, and compared to those of human tumor infiltrating Treg cells.ResultsIL-17 Receptor A (IL-17RA) is expressed in Tregs that reside in mouse mesenteric lymph nodes and colon tumors. Ablation of IL-17RA, specifically in Tregs, resulted in increased Th17 cells, and exacerbated tumor development. Mechanistically, tumor-infiltrating Tregs exhibit a unique gene signature that is linked to their activation, maturation, and suppression function, and this signature is in part supported by the direct signaling of IL-17 to Tregs. To study pathways of Treg programming, we found that loss of IL-17RA in tumor Tregs resulted in reduced RNA splicing, and downregulation of several RNA binding proteins that are known to regulate alternative splicing and promote Treg function.ConclusionIL-17 directly signals to Tregs and promotes their maturation and function. This signaling pathway constitutes a negative feedback loop that controls cancer-promoting inflammation in CRC.

With the discovery of evidence that many endocrine-disrupting chemicals (EDCs) in the environment influence human health, their toxic effects and mechanisms have become a hot topic of research. However, investigations into their endocrine-disrupting toxicity under combined binary exposure, especially the molecular mechanism of combined effects, have rarely been documented. In this study, two typical EDCs, perfluorooctanoic acid (PFOA) and 4-hydroxybenzophenone (4-HBP), were selected to examine their combined effects and molecular mechanism on MCF-7 cell proliferation at environmentally relevant exposure concentrations. We have successfully established a model to evaluate the binary combined toxic effects of endocrine disruptors, presenting combined effects in a simple and direct way. Results indicated that the combined effect changed from additive to synergistic from 1.25 × 10−8 M to 4 × 10−7 M. Metabolomics analyses suggested that exposure to PFOA and 4-HBP caused significant alterations in purine metabolism, arginine, and proline metabolism and had superimposed influences on metabolism. Enhanced combined effects were observed in glycine, serine, and threonine metabolic pathways compared to exposure to PFOS and 4-HBP alone. Additionally, the differentially expressed genes (DEGs) are primarily involved in Biological Processes, especially protein targeting the endoplasmic reticulum, and significantly impact the oxidative phosphorylation and thermogenesis-related KEGG pathway. By integrating metabolome and transcriptome analyses, PFOA and 4-HBP regulate purine metabolism, the TCA cycle, and endoplasmic reticulum protein synthesis in MCF-7 cells via mTORC1, which provides genetic material, protein, and energy for cell proliferation. Furthermore, molecular docking confirmed the ability of PFOA and 4-HBP to stably bind the estrogen receptor, indicating that they have different binding pockets. Collectively, these findings will offer new insights into understanding the mechanisms by which EDCs produce combined toxicity.

The Golgi apparatus plays a crucial role in mediating the modification, transport, and sorting of intracellular proteins and lipids. The morphological changes occurring in the Golgi apparatus are exceptionally important for maintaining its function. When exposed to external pressure or environmental stimulation, the Golgi apparatus undergoes adaptive changes in both structure and function, which are known as Golgi stress. Although certain signal pathway responses or post-translational modifications have been observed following Golgi stress, further research is needed to comprehensively summarize and understand the related mechanisms. Currently, there is evidence linking Golgi stress to neurodegenerative diseases; however, the role of Golgi stress in the progression of neurodegenerative diseases such as Alzheimer's disease remains largely unexplored. This review focuses on the structural and functional alterations of the Golgi apparatus during stress, elucidating potential mechanisms underlying the involvement of Golgi stress in regulating immunity, autophagy, and metabolic processes. Additionally, it highlights the pivotal role of Golgi stress as an early signaling event implicated in the pathogenesis and progression of neurodegenerative diseases. Furthermore, this study summarizes prospective targets that can be therapeutically exploited to mitigate neurodegenerative diseases by targeting Golgi stress. These findings provide a theoretical foundation for identifying novel breakthroughs in preventing and treating neurodegenerative diseases.

Lycium barbarum L., an important functional food in China, has antioxidant and antiaging activity. However, the exact antioxidant activity mechanism of Lycium barbarum extracts (LBE) is not well understood. Therefore, a carbendazim (CBZ)-induced PC12 cell injury model was constructed and vitrificated to study the antioxidant activity of fresh LBE on the basis of extraction parameter optimization via the full factorial design of experiments (DOE) method. The results showed that the pretreatment of PC12 cells with LBE could reduce the reactive oxygen species (ROS) level by 14.6% and inhibited the mitochondrial membrane potential (MMP) decline by 12.0%. Furthermore, the integrated analysis revealed that LBE played an antioxidant role by activating oxidative phosphorylation (OXPHOS) and restoring MMP, maintaining the tricarboxylic acid (TCA) cycle stability, and regulating the GSH metabolic pathway. The results of the present study provide new ideas for the understanding of the antioxidant function of LBE from a global perspective.

The antenna is an important part of the mine UWB positioning system. In order to suppress the multipath reflection signal received by the mine UWB positioning system and improve the positioning precision of the mine UWB positioning system, the traditional circularly polarized antenna structure is optimized. This study designs a new type of right-handed circularly polarized microstrip antenna. Firstly, the rectangular slot is changed to the cross slot. The current trajectory is changed to make the difference between the two orthogonal linearly polarized electric fields closer to 90°. The axial ratio is reduced to make the circularly polarized purity of the antenna higher. Secondly, the cross slot is smoothly expanded into a diamond slot to improve the cross-polarization level and suppress the orthogonal rotation. Therefore, the antenna can achieve higher right-handed circularly polarized purity. The HFSS software is used to simulate the new right-handed circularly polarized microstrip antenna. The simulation results show that the cross-polarization level in the main radiation direction is more than 10.4 dBi, which can achieve good right-handed circularly polarized effect. The measured results of the operating frequency band and echo of the new right-handed circularly polarized microstrip antenna sample are basically consistent with the simulation results. The results indicate that the new antenna structure is easy to be processed and the influence of the process deviation within the normal tolerance on the antenna performance is relatively small. The new right-handed circularly polarized microstrip antenna sample is used for the field test of the UWB positioning system based on the arrival phase difference in the tunnel environment. The test results show that the stability of the arrival phase difference of the signal is significantly improved. The multipath effect is significantly suppressed, which can effectively improve the positioning precision of the UWB positioning system. The new right-handed circularly polarized microstrip antenna has the advantages of small size, light weight and easy fabrication.

The primary objective of introducing metaheuristic algorithms into traditional systematic logic is to minimize the cost function. However, there is a lack of research on the impact of introducing metaheuristic algorithms on the cost function under different proportions of positive literals. In order to fill in this gap and improve the efficiency of the metaheuristic algorithm in systematic logic, we proposed a metaheuristic algorithm based on mutation tabu search and embedded it in probabilistic satisfiability logic in discrete Hopfield neural networks. Based on the traditional tabu search algorithm, the mutation operators of the genetic algorithm were combined to improve its global search ability during the learning phase and ensure that the cost function of the systematic logic converged to zero at different proportions of positive literals. Additionally, further optimization was carried out in the retrieval phase to enhance the diversity of solutions. Compared with nine other metaheuristic algorithms and exhaustive search algorithms, the proposed algorithm was superior to other algorithms in terms of time complexity and global convergence, and showed higher efficiency in the search solutions at the binary search space, consolidated the efficiency of systematic logic in the learning phase, and significantly improved the diversity of the global solution in the retrieval phase of systematic logic.

Background: Binafuxi granules are a traditional Uighur medicine (TUM) for treating the common cold with fever. However, high-quality clinical studies supporting its efficacy and safety are lacking. Methods: In this multicenter, randomized, double-blind, placebo-controlled, phase II clinical trial, patients with common cold and fever were randomly assigned to a high-dose group, low-dose group, and placebo group in a 1:1:1 ratio. Outcomes were time to fever relief, time to fever clearance, proportion of afebrile patients, time to symptom disappearance, rate of symptom disappearance, effective rate, emergency drug usage and safety assessment. Results: A total of 235 patients were recruited. Of these, 234 were included in the full analysis set (FAS), and 217 were included in the per-protocol set (PPS). In the FAS analysis, the median time to fever relief was 6.00 h, 5.54 h and 10.65 h (P = 0.31) in the high-dose group, low-dose group and placebo group, respectively. The median time to fever clearance was 18.29 h, 20.08 h and 25.00 h (P = 0.0018), respectively, and the proportion of afebrile patients was 92.4%, 89.7% and 71.4% (P = 0.0002), respectively. There was a significant difference in the disappearance time and disappearance rate of all symptoms and of individual symptoms. No serious adverse events were found. Conclusions: Binafuxi granules can dose-dependently shorten the fever course and improve clinical symptoms in patients suffering from the common cold with fever. Trial Registration: This trial was registered at Chinese Clinical Trial Registry (ChiCTR-IIR-17013379).

Dielectric materials with high energy storage density ([Formula: see text]) and efficiency ([Formula: see text]) are expected for energy storage capacitors. In this work, [Formula: see text]001[Formula: see text]-textured [Formula: see text][Formula: see text]NbO3 (NBN) ceramics were prepared by a templated grain growth technique. The effects of microstructure and orientation degree on dielectric properties, polarization and energy storage performance were investigated. The textured ceramic with an optimized orientation degree (70%) showed a high [Formula: see text] of 2.4 J/cm3 and [Formula: see text] of 85.6%. The excellent energy storage properties of textured ceramic originate from the co-effect of interfacial polarization and clamping effect. The results indicate that texture development is a potential candidate to optimize the energy storage properties of functional ceramics.

Anthracyclines, such as doxorubicin (Dox), are widely used chemotherapeutic agents for the treatment of solid tumors and hematologic malignancies. However, they frequently induce cardiotoxicity leading to dilated cardiomyopathy and heart failure. This study sought to investigate the role of the exchange protein directly activated by cAMP (EPAC) in Dox-induced cardiotoxicity and the potential cardioprotective effects of EPAC inhibition. We show that Dox induces DNA damage and cardiomyocyte cell death with apoptotic features. Dox also led to an increase in both cAMP concentration and EPAC1 activity. The pharmacological inhibition of EPAC1 (with CE3F4) but not EPAC2 alleviated the whole Dox-induced pattern of alterations. When administered in vivo, Dox-treated WT mice developed a dilated cardiomyopathy which was totally prevented in EPAC1 knock-out (KO) mice. Moreover, EPAC1 inhibition potentiated Dox-induced cell death in several human cancer cell lines. Thus, EPAC1 inhibition appears as a potential therapeutic strategy to limit Dox-induced cardiomyopathy without interfering with its antitumoral activity.

Abstract Background Cardiac myxofibrosarcoma is a rare cardiac malignant tumor, whose diagnosis is challenging due to its rare and non-specific manifestations. Ultrasound is the most important tool for detecting cardiac tumors. Yet, its diagnostic value in cardiac myxoidfibrosarcoma is rarely reported. Herein, we summarized the ultrasonic manifestations of myxofibrosarcoma in a 72-year-old Han woman. Case presentation The patient presented with crushing chest pain without obvious inducement, lasting 3–5 min each time, which would be relieved after rest, accompanied by palpitation, chest tightness, shortness of breath, dizziness, and syncope. The electrocardiogram (ECG) suggested atrial fibrillation. Ultrasound found two moderate echogenic masses in the left atrium; one was about 48 × 31 mm in size, and the other was about 25 × 24 mm in size. The clinical diagnosis was atrial mass and atrial fibrillation. The patient underwent the operation of left atrial tumor resection + mitral valvuloplasty + atrial fibrillation radiofrequency ablation + left atrial appendectomy. The tumor was completely removed, and the patient did not receive radiotherapy or chemotherapy after surgery. The patient was reexamined by ultrasound at 6, 42, and 91 days after surgery, and no obvious abnormalities were found. On day 115, moderate echoic mass was detected on the posterior wall of the left atrium, nearing the mitral valve ring, with a size of about 28 × 23 mm. Currently, the patient is under follow-up care. Conclusion As the most important examination method for cardiac tumors, cardiac ultrasound has good diagnostic and differential diagnosis value and can be used regularly due to its simplicity and safety. The diagnosis rate of cardiac myxofibrosarcoma can be greatly improved by summarizing the ultrasonographic manifestations of cardiac myxofibrosarcoma and differentiating them from other lesions.

How and whether histone modifications regulate distinct gene networks remains insufficiently understood. Here Cattaneo et al show that DOT1L catalyzed H3K79me2 regulates fetal chamber-specific gene expression and neonatal cardiomyocyte cell cycle withdrawal to coordinate heart development.

Abstract The winter South China Sea (SCS) western boundary current (WBC) has presented a significant weak–strong–weak phase shift in the last two decades, as revealed by observations and model products. Its phase shift has mainly been attributed to the joint effect of wind stress curl over the SCS and Luzon Strait transport (LST). Changes in the wind stress curl and planetary vorticity input through the Luzon Strait are the two major sources maintaining the winter SCS WBC state in different phases. The LST is closely related to the Kuroshio strength and thus the Pacific North Equatorial Current (NEC) bifurcation. Therefore, the winter SCS WBC phase shift is attributed to phase shifts of the winter monsoon and the NEC bifurcation, which are mostly related to those of the Indian Ocean Dipole Mode and Atlantic Multidecadal Oscillation, respectively. Understanding the SCS circulation low‐frequency variability is essential to forecast its thermodynamic state and regional climate effect.

FLNC, encoding filamin C, is one of the most mutated genes in dilated and hypertrophic cardiomyopathy. However, the precise role of filamin C in mammalian heart remains unclear. In this study, we demonstrated Flnc global (FlncgKO) and cardiomyocyte-specific knockout (FlnccKO) mice died in utero from severely ruptured ventricular myocardium, indicating filamin C is required to maintain the structural integrity of myocardium in the mammalian heart. Contrary to the common belief that filamin C acts as an integrin inactivator, we observed attenuated activation of β1 integrin specifically in the myocardium of FlncgKO mice. Although deleting β1 integrin from cardiomyocytes did not recapitulate the heart rupture phenotype in Flnc knockout mice, deleting both β1 integrin and filamin C from cardiomyocytes resulted in much more severe heart ruptures than deleting filamin C alone. Our results demonstrated that filamin C works in concert with β1 integrin to maintain the structural integrity of myocardium during mammalian heart development.

Background and Purpose: The aim of this study was to explore the effect of half a year of evolocumab plus moderate-intensity statin treatment on carotid intraplaque neovascularization (IPN) and blood lipid levels.Methods: A total of 31 patients with 33 carotid plaques who received evolocumab plus statin treatment were included. Blood lipid levels, B-mode ultrasound and contrast-enhanced ultrasonography (CEUS) at baseline and after half a year of evolocumab plus statin therapy were collected. The area under the curve (AUC) reflected the total amount of acoustic developer entering the plaque or lumen within the 180 s measurement period. The enhanced intensity reflected the peak blood flow intensity during the monitoring period, and the contrast agent area reflected the area of vessels in the plaques.Results: Except for high-density lipoprotein cholesterol (HDL-c), all other lipid indices decreased. Compared with baseline, low-density lipoprotein cholesterol (LDL-c) decreased by approximately 57% (p < 0.001); total cholesterol (TC) decreased by approximately 34% (p < 0.001); small dense low-density lipoprotein (sd-LDL) decreased by approximately 52% (p < 0.001); and HDL-c increased by approximately 20% (p < 0.001). B-mode ultrasonography showed that the length and thickness of the plaque and the hypoechoic area ratio were reduced (p < 0.05). The plaque area, calcified area ratio, and lumen cross-sectional area changed little (p > 0.05). CEUS revealed that the area under the curve of plaque/lumen [AUC (P/L)] decreased from 0.27 ± 0.13 to 0.19 ± 0.11 (p < 0.001). The enhanced intensity ratio of plaque/lumen [intensity ratio (P/L)] decreased from 0.37 ± 0.16 to 0.31 ± 0.14 (p = 0.009). The contrast agent area in plaque/area of plaque decreased from 19.20 ± 13.23 to 12.66 ± 9.59 (p = 0.003). The neovascularization score decreased from 2.64 ± 0.54 to 2.06 ± 0.86 (p < 0.001). Subgroup analysis based on statin duration (

BackgroundAs increasing experimental evidence suggests that iron metabolism play crucial roles in cancer and non-cancer conditions, there is a lack of data on serum soluble transferrin receptor (sTfR), a promising marker representing unmet cellular iron demands, between cancer risk from epidemiological studies. Here, we aimed to evaluate the predictive value of sTfR and cancer prevalence.Materials and methodsWe analyzed on 5,480 adult participants from 2015 to 2018 National Health and Nutrition Examination Survey (NHANES). Spearman correlation analysis was performed to investigate the correlations between sTfR and other characteristics. To identify the associations between sTfR and the prevalence of cancers, stratified multivariable logistic regression models, subgroup and sensitivity analyses were also performed.ResultsIn tertile analyses, participants in the highest level of sTfR were significantly associated with increased prevalence of total cancers [odds ratio (OR) = 1.53, 95% confidence interval (CI): 1.15-2.02] as compared with those at the lowest tertile. Each unit increment in ln-transformed sTfR concentration was shown to be associated with 39% increased risks of total cancers. Similar associations were found in males rather than females. Further subgroup and sensitivity analyses indicated that, in continuous and tertile analyses, sTfR was more closely associated with male- and female-specific cancers of prostate and testis (2.35: 1.03-5.40; 2.03: 1.00-4.09; respectively), and breast, cervix, ovary and uterus (1.92: 1.11-3.35; 1.66: 1.02-2.69; respectively).ConclusionsOur findings suggested that elevated level of sTfR was associated with the prevalence of cancers, especially in sex-specific cancers. In order to better determine them, further research in humans will be required.

In this paper, a series of glycyrrhetic acid derivatives 3a–3f were synthesized via the esterification reaction. The cytotoxicity of these compounds against five tumor cells (SGC-7901, BEL-7402, A549, HeLa and B16) and normal LO2 cells was investigated using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The results showed that compound 3a exhibited high antiproliferative activity against HeLa cells (IC50 = 11.4 ± 0.2 μM). The anticancer activity was studied through apoptosis, cloning, and scratching; the levels of the intracellular ROS, GSH, and Ca2+; and the change in the mitochondrial membrane potential, cell cycle arrest and RNA sequencing. Furthermore, the effects of compound 3a on gene expression levels and metabolic pathways in HeLa cells were investigated via transcriptomics. The experimental results showed that this compound can block the cell cycle in the S phase and inhibit cell migration by downregulating Focal adhesion kinase (FAK) expression. Moreover, the compound can reduce the intracellular glutathione (GSH) content, increase the Ca2+ level and the intracellular ROS content, and induce a decrease in the mitochondrial membrane potential, further leading to cell death. In addition, it was also found that the mechanism of compounds inducing apoptosis was related to the regulation of the expression of mitochondria-related proteins B-cell lymphoma-2 (Bcl-2), Bcl-2-Associated X (Bax), and the activation of the caspase proteins. Taken together, this work provides a help for the development of glycyrrhetinic acid compounds as potential anticancer molecules.

BackgroundNowadays, intelligent medicine is gaining widespread attention, and great progress has been made in Western medicine with the help of artificial intelligence to assist in decision making. Compared with Western medicine, traditional Chinese medicine (TCM) involves selecting the specific treatment method, prescription, and medication based on the dialectical results of each patient’s symptoms. For this reason, the development of a TCM-assisted decision-making system has lagged. Treatment based on syndrome differentiation is the core of TCM treatment; TCM doctors can dialectically classify diseases according to patients’ symptoms and optimize treatment in time. Therefore, the essence of a TCM-assisted decision-making system is a TCM intelligent, dialectical algorithm. Symptoms stored in electronic medical records are mostly associated with patients’ diseases; however, symptoms of TCM are mostly subjectively identified. In general electronic medical records, there are many missing values. TCM medical records, in which symptoms tend to cause high-dimensional sparse data, reduce algorithm accuracy. ObjectiveThis study aims to construct an algorithm model compatible for the multidimensional, highly sparse, and multiclassification task of TCM syndrome differentiation, so that it can be effectively applied to the intelligent dialectic of different diseases. MethodsThe relevant terms in electronic medical records were standardized with respect to symptoms and evidence-based criteria of TCM. We structuralized case data based on the classification of different symptoms and physical signs according to the 4 diagnostic examinations in TCM diagnosis. A novel cross-feature generation by convolution neural network model performed evidence-based recommendations based on the input embedded, structured medical record data. ResultsThe data set included 5273 real dysmenorrhea cases from the Sichuan TCM big data management platform and the Chinese literature database, which were embedded into 60 fields after being structured and standardized. The training set and test set were randomly constructed in a ratio of 3:1. For the classification of different syndrome types, compared with 6 traditional, intelligent dialectical models and 3 click-through-rate models, the new model showed a good generalization ability and good classification effect. The comprehensive accuracy rate reached 96.21%. ConclusionsThe main contribution of this study is the construction of a new intelligent dialectical model combining the characteristics of TCM by treating intelligent dialectics as a high-dimensional sparse vector classification task. Owing to the standardization of the input symptoms, all the common symptoms of TCM are covered, and the model can differentiate the symptoms with a variety of missing values. Therefore, with the continuous improvement of disease data sets, this model has the potential to be applied to the dialectical classification of different diseases in TCM.

Adopting proton exchange membrane fuel cells fuelled by hydrogen presents a promising solution for the shipping industry’s deep decarbonisation. However, the potential safety risks associated with hydrogen leakage pose a significant challenge to the development of hydrogen-powered ships. This study examines the safe design principles and leakage risks of the hydrogen gas supply system of China’s first newbuilt hydrogen-powered ship. This study utilises the computational fluid dynamics tool FLACS to analyse the hydrogen dispersion behaviour and concentration distributions in the hydrogen fuel cell room based on the ship’s parameters. This study predicts the flammable gas cloud and time points when gas monitoring points first reach the hydrogen volume concentrations of 0.8% and 1.6% in various leakage scenarios, including four different diameters (1, 3, 5, and 10 mm) and five different directions. This study’s findings indicate that smaller hydrogen pipeline diameters contribute to increased hydrogen safety. Specifically, in the hydrogen fuel cell room, a single-point leakage in a hydrogen pipeline with an inner diameter not exceeding 3 mm eliminates the possibility of flammable gas cloud explosions. Following a 10 mm leakage diameter, the hydrogen concentration in nearly all room positions reaches 4.0% within 6 s of leakage. While the leakage diameter does not impact the location of the monitoring point that first activates the hydrogen leak alarm and triggers an emergency hydrogen supply shutdown, the presence of obstructions near hydrogen detectors and the leakage direction can affect it. These insights provide guidance on the optimal locations for hydrogen detectors in the fuel cell room and the pipeline diameters on hydrogen gas supply systems, which can facilitate the safe design of hydrogen-powered ships.

In order to study the axial compression behavior of magnetically-driven concrete (MDC) filled double-skin GFRP tubes (CFDGT), the axial compression test was conducted on eight columns (the length of columns ranged from 1,000 mm to 3,000 mm). For the MDC, a magnetic vibration device was established to vibrate the MDC, and two series of MDC (concrete with and without vibration) were adopted to investigate the vibration effect, research results showed that the concrete between two circular GFRP tubes could be effectively vibrated by the magnetic force. For the CFDGT columns with MDC, the load-displacement curves, load-strain curves, failure modes and ultimate loads of tested columns were obtained. After the axial compression test, a finite element model was proposed to predict the axial compression behavior and carry out parametric analysis. The design equations for the ultimate load of CFDGTs were derived from the finite element results, comparison results showed that the design equations can predict the ultimate load of CFDGT columns accurately and conservatively, and the maximum calculation error was controlled within 10%.

BackgroundGlucose variability (GV) is a common complication of dysglycemia in critically ill patients. However, there are few studies on the role of GV in the prognosis of pediatric patients, and there is no consensus on the appropriate method for GV measurement. The objective of this study was to determine the “optimal” index of GV in non-diabetic critically ill children in a prospective multicenter cohort observational study. Also, we aimed to confirm the potential association between GV and unfavorable outcomes and whether this association persists after controlling for hypoglycemia or hyperglycemia.Materials and MethodsBlood glucose values were recorded for the first 72 h and were used to calculate the GV for each participant. Four different metrics [SD, glycemic lability index (GLI), mean absolute glucose (MAG), and absolute change of percentage (ACACP)] were considered and compared to identify the “best” GV index associated with poor prognosis in non-diabetic critically ill children. Among the four metrics, the SD was most commonly used in previous studies, while GLI- and MAG-integrated temporal information, that is the rate and magnitude of change and the time interval between glucose measurements. The fourth metric, the average consecutive ACACP, was introduced in our study, which can be used in real-time clinical decisions. The primary outcome of this study was the 28-day mortality. The receiver operating characteristic (ROC) curve analysis was conducted to compare the predictive power of different metrics of GV for the primary outcome. The GV index with the largest area under ROC curve (AUC) was chosen for subsequent multivariate analyses. Multivariate Cox regression analysis was performed to identify the potential predictors of the outcome. To compare the contribution in 28-day mortality prognosis between glycemic variability and hyper- or hypoglycemia, performance metrics were calculated, which included AUC, net reclassification improvement (NRI), and integrated discrimination improvement (IDI).ResultsAmong 780 participants, 12.4% (n = 97) died within 28 days after admission to the pediatric intensive care unit (PICU). Statistically significant differences were found between survivors and non-survivors in terms of four GV metrics (SD, GLI, MAG, and ACACP), in which MAG (AUC: 0.762, 95% CI: 0.705–0.819, p < 0.001) achieved the largest AUC and showed a strong independent association with ICU mortality. Subsequent addition of MAG to the multivariate Cox model for hyperglycemia resulted in further quantitative evolution of the model statistics (AUC = 0.651–0.681, p = 0.001; IDI: 0.017, p = 0.044; NRI: 0.224, p = 0.186). The impact of hyperglycemia (adjusted hazard ratio [aHR]: 1.419, 95% CI: 0.815–2.471, p = 0.216) on outcome was attenuated and no longer statistically relevant after adjustment for MAG (aHR: 2.455, 95% CI: 1.411–4.270, p = 0.001).ConclusionsGV is strongly associated with poor prognosis independent of mean glucose level, demonstrating more predictive power compared with hypoglycemia and hyperglycemia after adjusting for confounding factors. GV metrics that contain information, such as time and rate of change, are the focus of future research; thus, the MAG may be a good choice. The findings of this study emphasize the crucial role of GVs in children in the PICU. Clinicians should pay more attention to GV for clinical glucose management.

In view of problems in existing rolling bearing fault diagnosis methods for coal mine rotating machinery, such as incomplete signal feature extraction, low fault diagnosis accuracy and low efficiency, a rolling bearing fault diagnosis method based on wavelet packet decomposition and particle swarm optimization BP neural network was proposed. The method includes signal feature extraction and fault type recognition. In the signal feature extraction part, the collected vibration signals of rolling bearing are decomposed by wavelet packet to obtain energy of each sub-frequency band and total energy of the signal. After normalization processing, feature vector representing state of rolling bearing is obtained. In the fault type recognition part, initial weight and threshold of BP neural network are optimized by particle swarm optimization to accelerate convergence speed of the network and avoid falling into local minimum. The experimental results show that the method improves fault diagnosis efficiency and accuracy of rolling bearing.

Abstract QPOs are seen as peak features in the X-ray power spectral density of stellar mass black holes and neutron stars, of which frequencies can be measured with high precision. These QPO frequencies are believed to be related to fundamental frequencies of test particles, which are mainly determined by the background metric. We consider the metric introduced in Ghasemi-Nodehi and Bambi (Eur Phys J C 76:290, 2016). The fundamental frequencies in this metric are sensitive to some of the metric parameters but insensitive to other parameters, which means, the differences in fundamental frequencies in this metric and those in the Kerr ones can be significant for small changes of some but not all parameters around the Kerr value. By comparing with the QPO observations of GRO J1655-40, we find that only one parameter of the Ghasemi-Nodehi–Bambi metric can be strongly constrained, but other parameters cannot. We also use nested algorithm to investigate whether better constraints on the Ghasemi-Nodehi–Bambi metric parameters can be obtained from QPO observations of multiple objects by simulations. We find that four parameters can be strongly constrained while other parameters cannot . Our results suggest that QPOs may be important tools for testing the Kerr metric.

Abstract Compelling evidence has demonstrated the potential functions of circular RNAs (circRNAs) in breast cancer (BC) tumorigenesis. Nevertheless, the underlying mechanism by which circRNAs regulate BC progression is still unclear. The purpose of present research was to investigate the novel circRNA circRNF20 (hsa_circ_0087784) and its role in BC. CircRNA microarray sequencing revealed that circRNF20 was one of the upregulated transcripts in BC samples. Increased circRNF20 level predicted the poor clinical outcome in BC specimens. Functionally, circRNF20 promoted the proliferation and Warburg effect (aerobic glycolysis) of BC cells. Mechanistically, circRNF20 harbor miR-487a, acting as miRNA sponge, and then miR-487a targeted the 3’-UTR of hypoxia-inducible factor-1α (HIF-1α). Moreover, HIF-1α could bind with the promoter of hexokinase II (HK2) and promoted its transcription. In conclusion, this finding illustrates the vital roles of circRNF20 via the circRNF20/ miR-487a/HIF-1α/HK2 axis in breast cancer progress and Warburg effect, providing an interesting insight for the BC tumorigenesis.

Abstract Background The IL-17 family cytokines are potent drivers of colorectal cancer (CRC) development. We and others have shown that IL-17 mainly signals to tumor cells to promote CRC, but the underlying mechanism remains unclear. IL-17 also dampens Th1-armed anti-tumor immunity, in part by attracting myeloid cells to tumor. Whether IL-17 controls the activity of adaptive immune cells in a more direct manner, however, is unknown. Methods Using mouse models of sporadic or inducible colorectal cancers, we ablated IL-17RA in the whole body or specifically in colorectal tumor cells. We also performed adoptive bone marrow reconstitution to knockout CXCR3 in hematopoietic cells. Histological and immunological experimental methods were used to reveal the link among IL-17, chemokine production, and CRC development. Results Loss of IL-17 signaling in mouse CRC resulted in marked increase in the recruitment of CD8+ cytotoxic T lymphocytes (CTLs) and regulatory T cells (Tregs), starting from early stage CRC lesions. This is accompanied by the increased expression of anti-inflammatory cytokines IL-10 and TGF-β. IL-17 signaling also inhibits the production of T cell attracting chemokines CXCL9 and CXCL10 by tumor cells. Conversely, the inability of hematopoietic cells to respond to CXCL9/10 resulted in decreased tumor infiltration by CTLs and Tregs, decreased levels of IL-10 and TGF-β, and increased numbers of tumor lesions. Blockade of IL-17 signaling resulted in increased expression of immune checkpoint markers. On the other hand, treatment of mouse CRC with anti-CTLA-4 antibody led to increased expression of pro-tumor IL-17. Conclusion IL-17 signals to colorectal tumor cells and inhibits their production of CXCL9/10 chemokines. By doing so, IL-17 inhibits the infiltration of CD8+ CTLs and Tregs to CRC, thus promoting CRC development. Cancer immunotherapy may be benefited by the use of anti-IL-17 agents as adjuvant therapies, which serve to block both IL-17-mediated tumor promotion and T cell exclusion.

This study reveals the features of the strong intraseasonal variability (ISV) of the upper-layer current in the northern South China Sea (NSCS) based on four long-time mooring observations and altimeter data. The ISV of the upper-layer current in the NSCS consists of two dominant periods of 10–65 days and 65–110 days. The ISV with period of 10–65 days is much strong in the Luzon Strait and decays rapidly westward along the slope. The ISV with the period of 65–110 days is relatively strong along the slope with two high cores at 115 and 119°E, whereas it is weak in the Luzon Strait. The 10–65-day ISV can propagate directly from the western Pacific into the NSCS for most of the time. However, due to its long wavelength, the 65–110-day ISV propagates into the NSCS indirectly, possibly similar to the wave diffraction phenomenon. The spatial differences between the two main frequency bands are primarily due to the baroclinic and barotropic instabilities. The spatial distribution of the upper-layer ISV is closely associated with the mesoscale eddy radius of the NSCS. The eddy radius is directly proportional to the strength of 65–110-day ISV, but it is inversely proportional to the strength of 10–65-day ISV.

To examine the spatial pattern and controlling factors of the primary productivity (PP) of phytoplankton in the eastern Indian Ocean (EIO), deck-incubation carbon fixation (a 14C tracer technique) and the related hydrographic properties were measured at 15 locations during the pre-summer monsoon season (February–April 2017). There are knowledge gaps in the field observations of PP in the EIO. The estimated daily carbon production rates integrated over the photic zone ranged from 113 to 817 mgC m–2 d–1, with a mean of 522 mgC m–2 d–1. The mixed-layer integrated primary production (MLD-PP) ranged from 29.0 to 303.7 mgC m–2 d–1 (mean: 177.2 mgC m–2 d–1). The contribution of MLD-PP to the photic zone-integrated PP (PZI-PP) varied between 19 and 51% (mean: 36%). Strong spatial variability in the carbon fixation rates was found in the study region. Specifically, the surface primary production rates were relatively higher in the Bay of Bengal domain affected by riverine flux and lower in the equatorial domain owing to the presence of intermonsoonal Wyrtki jets, which were characterized by a depression of thermocline and nitracline. The PZI-PP exhibited a linear (positive) relationship with nutrient values, but with no significance, indicating a partial control of macronutrients and a light limitation of carbon fixation. As evident from the vertical profiles, the primary production process mainly occurred above the nitracline depth and at high photosynthetic efficiency. Phytoplankton (>5 μm), including dinoflagellates, Trichodesmium, coccolithophores, and dissolved nutrients, are thought to have been correlated with primary production during the study period. The measured on-deck biological data of our study allow for a general understanding of the trends in PP in the survey area of the EIO and can be incorporated into global primary production models.

In this study, the dose-dependent beneficial effects of tuna oil (TO) on cigarette smoke exposure induced lung inflammation were investigated. Treatments with low- (LD), moderate- (MD) and high-dosage (HD) of TO reduce the C-reactive protein concentration in serum, significantly down-regulated and up-regulated the mRNA levels of proinflammatory and anti-inflammatory cytokines in the lung, respectively. TO treatment inhibited the activation of the nuclear factor-kappaB (NF-κB) signaling pathway and the NLR family pyrin domain-containing 3 (NLRP3) inflammasome. In addition, TO treatments reduced the MDA concentration and enhanced the activities of SOD and GSH-Px in serum. What’s more, TO treatment modulated the gut microbiota by increasing the abundance of Lactobacillus species and short-chain fatty acid producers and decreasing the abundance of opportunistic pathogens, as well as affecting the amino acid metabolism in serum. This research clarified the anti-inflammatory mechanism of TO on cigarette smoking exposure induced lung inflammation in mice.

The sine and cosine algorithm is a new simple and effective population optimization method proposed in recent years that has been studied in many works of literature. Based on the basic principle of the sine and cosine algorithm, this paper fully studies the main parameters affecting the performance of the sine and cosine algorithm, integrates the reverse learning algorithm, adds an elite opposition solution and forms the hybrid sine and cosine algorithm (hybrid SCA). Combined with the fuzzy k-nearest neighbor method and the hybrid SCA, this paper numerically simulates two-class datasets and multi-class datasets, obtains a large number of numerical results and analyzes the results. The hybrid SCA FKNN proposed in this paper has achieved good accuracy in classification and prediction results under 10 different types of data sets. Compared with SCA FKNN, LSCA FKNN, BA FKNN, PSO FKNN and SSA FKNN, the prediction accuracy is significantly improved. In the Wilcoxon signed rank test with SCA FKNN and LSCA FKNN, the zero hypothesis (significance level 0.05) is rejected and the two classifiers have a significantly different accuracy.

Mammotome-an ultrasound guided vacuum-assisted breast biopsy (VABB) device, has proved beneficial to the treatment of benign breast lesions. The aim of this study is to analyze the characteristics of ultrasound images of residual cavity and the changes in ultrasound images at follow-up at different intervals after the excision of benign breast lesions by Mammotome® biopsy system. A series of 247 consecutive 8-gauge Mammotome® procedures were performed under ultrasound guidance and multivariate analysis was conducted. We found fibroadenoma and adenomatosis are appeared to be the most common pathological manifestations. Follow-up by ultrasonography at an interval of one month after excision of benign breast lesions by 8-gauge vacuum-assisted Mammotome® biopsy system, is not reliable due to the residual cavity formation. A follow-up schedule starting from at least 3 months after resection is highly recommended. Keywords: Mammotome, Benign breast lesion, Ultrasound image, Residual cavity, Echogenicity

One of the main problems in representing information in the form of nonsystematic logic is the lack of flexibility, which leads to potential overfitting. Although nonsystematic logic improves the representation of the conventional k Satisfiability, the formulations of the first, second, and third-order logical structures are very predictable. This paper proposed a novel higher-order logical structure, named G-Type Random k Satisfiability, by capitalizing the new random feature of the first, second, and third-order clauses. The proposed logic was implemented into the Discrete Hopfield Neural Network as a symbolic logical rule. The proposed logic in Discrete Hopfield Neural Networks was evaluated using different parameter settings, such as different orders of clauses, different proportions between positive and negative literals, relaxation, and differing numbers of learning trials. Each evaluation utilized various performance metrics, such as learning error, testing error, weight error, energy analysis, and similarity analysis. In addition, the flexibility of the proposed logic was compared with current state-of-the-art logic rules. Based on the simulation, the proposed logic was reported to be more flexible, and produced higher solution diversity.

The behavior of two series of corroded thin-walled concrete-filled steel tubes (CFSTs) was investigated in this paper. One series, called the CT series, consists of fourteen CFST specimens corroded by NaCl solution, while the other, called the AT series, consists of six test pieces corroded in the atmosphere. Outer surfaces of corroded steel tubes were scanned using a 3D scanner machine, and a complete surface profile of each specimen was obtained for corrosion evaluation using different methods in this study. Entire test pieces were compressed to failure, and their behaviors under loading were recorded. It is shown that corrosion has a fractal feature, and it has a significant effect on the ductility of specimens. In addition, FEA models of corroded CFST specimens were established based on the 3D scanning profile results using the ABAQUS program and verified against column test results.

Atmospheric deposition is the dominant pathway for the loading of exogenous nitrogen (N) to open ocean. Here, rainwater samples were collected from 31 stations in the equatorial East Indian Ocean (EIO) and West Pacific Ocean (WPO) to explore the spatial variability of N species, potential sources, and related ecological influences. Among two oceans, nitrate (NO3–) and ammonium (NH4+) were the main components in the rainwater N inventory. NO3– concentrations varied from 0.19 to 100.5 μM, whereas NH4+ concentrations ranged from 0.54 to 110.6 μM. Among all stations, low concentrations of NO3– and NH4+ appeared in the remote ocean, whereas high concentrations were observed at the stations near the Malacca Strait and New Guinea, coupled with an enhancement of non-sea salt major ions, e.g., calcium ions (Ca2+) and sulfate (SO42–), revealing the influence from coastal human activities, such as coal and gasoline combustion. In the remote ocean, δ15N–NH4+ ranged from −5.7 to −9.3‰, whereas it dropped to -15.5‰ near coasts. A logarithmic decay between δ15N–NH4+ and NH4+ concentrations in rainwater samples was obtained, suggesting a shift from natural source (seawater emission) in oceanic precipitation events to anthropogenic source (chemical fertilizer volatilization and vehicle exhaust) in coastal rainwaters. δ15N–NO3– in the remote ocean varied between −1.7 and 0.4‰ with low levels found in the WPO, likely related to the ascending air flow driven by the Walker Circulation. In coastal oceans, δ15N–NO3– ranged from 1.5 to 3.5‰. The linkage between δ15N–NO3– and NO3– concentrations varied in two oceans, resulting from difference in biological and fossil fuel combustion contributions. Compared with ocean surface water, N in the rainwater was markedly enriched, suggesting that N from atmospheric wet depositions could rapidly enhance the dissolved N availability in ocean surface water. However, the N redundancy according to the Redfield–Brzezinski ratio (N:Si:P = 16:16:1) in the rainwater might benefit from the growth of N-preference phytoplankton species and microbes. As the first study on N concentrations, sources, and stoichiometry balance in rainwater over the equatorial WPO and EIO, the results could be a support to the global N budget estimation and oceanic primary production modeling.

Three benzoxanthone derivatives were synthesized through a new photochemical strategy. The in vitro cytotoxic activity of these compounds was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and their partition coefficients (logP) were measured by shake flask method. The pKa values of the compounds were detected by potentionmetric titration. The interaction of the compounds with calf thymus DNA (CT-DNA) was investigated by electronic absorption, luminescence spectra and viscosity. A molecular docking analysis was performed. The antitumor efficacy of the compounds was evaluated by cell apoptosis, cell cycle arrest, intracellular Ca2+ concentrations and reactive oxygen species (ROS) levels. The mitochondrial membrane potential was assayed using JC-1 (5,5′,6,6′-tetrachloro-1,1,3′,3′-tetraethyl-imidacarbocyanine iodide) as the fluorescence probe. The expression of Bcl-2 family protein, caspase 3 and poly ADP-ribose polymerase (PARP) was explored by western blot. The results showed that the compounds induced apoptosis through a ROS-mediated mitochondrial dysfunction pathway. This work provides an efficient approach to synthesize benzoxanthone derivatives, and is helpful for understanding the apoptotic mechanism.

Dilated cardiomyopathy (DCM) is a life-threatening form of heart disease that is typically characterized by progressive thinning of the ventricular walls, chamber dilation, and systolic dysfunction. Multiple mutations in the gene encoding filamin C (FLNC), an actin-binding cytoskeletal protein in cardiomyocytes, have been found in patients with DCM. However, the mechanisms that lead to contractile impairment and DCM in patients with FLNC variants are poorly understood. To determine how FLNC regulates systolic force transmission and DCM remodeling, we used an inducible, cardiac-specific FLNC-knockout (icKO) model to produce a rapid onset of DCM in adult mice. Loss of FLNC reduced systolic force development in single cardiomyocytes and isolated papillary muscles but did not affect twitch kinetics or calcium transients. Electron and immunofluorescence microscopy showed significant defects in Z-disk alignment in icKO mice and altered myofilament lattice geometry. Moreover, a loss of FLNC induces a softening myocyte cortex and structural adaptations at the subcellular level that contribute to disrupted longitudinal force production during contraction. Spatially explicit computational models showed that these structural defects could be explained by a loss of inter-myofibril elastic coupling at the Z-disk. Our work identifies FLNC as a key regulator of the multiscale ultrastructure of cardiomyocytes and therefore plays an important role in maintaining systolic mechanotransmission pathways, the dysfunction of which may be key in driving progressive DCM.

Nexilin (NEXN) was recently identified as a component of the junctional membrane complex required for development and maintenance of cardiac T-tubules. Loss of Nexn in mice leads to a rapidly progressive dilated cardiomyopathy (DCM) and premature death. A 3 bp deletion (1948–1950del) leading to loss of the glycine in position 650 (G650del) is classified as a variant of uncertain significance in humans and may function as an intermediate risk allele. To determine the effect of the G650del variant on cardiac structure and function, we generated a G645del-knockin (G645del is equivalent to human G650del) mouse model. Homozygous G645del mice express about 30% of the Nexn expressed by WT controls and exhibited a progressive DCM characterized by reduced T-tubule formation, with disorganization of the transverse-axial tubular system. On the other hand, heterozygous Nexn global KO mice and genetically engineered mice encoding a truncated Nexn missing the first N-terminal actin-binding domain exhibited normal cardiac function, despite expressing only 50% and 20% of the Nexn, respectively, expressed by WT controls, suggesting that not only quantity but also quality of Nexn is necessary for a proper function. These findings demonstrated that Nexn G645 is crucial for Nexn’s function in tubular system organization and normal cardiac function.

O-linked N-acetylglucosamine (GlcNAc) transferase (OGT) is the only enzyme catalyzing O-GlcNAcylation. Although it has been shown that OGT plays an essential role in maintaining postnatal heart function, its role in heart development remains unknown. Here we showed that loss of OGT in early fetal cardiomyocytes led to multiple heart developmental defects including hypertrabeculation, biventricular dilation, atrial septal defects, ventricular septal defects, and defects in coronary vessel development. In addition, RNA sequencing revealed that Angiopoietin-1, required within cardiomyocytes for both myocardial and coronary vessel development, was dramatically downregulated in cardiomyocyte-specific OGT knockout mouse hearts. In conclusion, our data demonstrated that OGT plays an essential role in regulating heart development through activating expression of cardiomyocyte Angiopoietin-1.

Inositol 1,4,5-trisphosphate receptors (IP3Rs) are a family of intracellular Ca2+ release channels located on the ER membrane, which in mammals consist of 3 different subtypes (IP3R1, IP3R2, and IP3R3) encoded by 3 genes, Itpr1, Itpr2, and Itpr3, respectively. Studies utilizing genetic knockout mouse models have demonstrated that IP3Rs are essential for embryonic survival in a redundant manner. Deletion of both IP3R1 and IP3R2 has been shown to cause cardiovascular defects and embryonic lethality. However, it remains unknown which cell types account for the cardiovascular defects in IP3R1 and IP3R2 double knockout (DKO) mice. In this study, we generated conditional IP3R1 and IP3R2 knockout mouse models with both genes deleted in specific cardiovascular cell lineages. Our results revealed that deletion of IP3R1 and IP3R2 in cardiomyocytes by TnT-Cre, in endothelial / hematopoietic cells by Tie2-Cre and Flk1-Cre, or in early precursors of the cardiovascular lineages by Mesp1-Cre, resulted in no phenotypes. This demonstrated that deletion of both IP3R genes in cardiovascular cell lineages cannot account for the cardiovascular defects and embryonic lethality observed in DKO mice. We then revisited and performed more detailed phenotypic analysis in DKO embryos, and found that DKO embryos developed cardiovascular defects including reduced size of aortas, enlarged cardiac chambers, as well as growth retardation at embryonic day (E) 9.5, but in varied degrees of severity. Interestingly, we also observed allantoic-placental defects including reduced sizes of umbilical vessels and reduced depth of placental labyrinth in DKO embryos, which could occur independently from other phenotypes in DKO embryos even without obvious growth retardation. Furthermore, deletion of both IP3R1 and IP3R2 by the epiblast-specific Meox2-Cre, which targets all the fetal tissues and extraembryonic mesoderm but not extraembryonic trophoblast cells, also resulted in embryonic lethality and similar allantoic-placental defects. Taken together, our results demonstrated that IP3R1 and IP3R2 play an essential and redundant role in maintaining the integrity of fetal-maternal connection and embryonic viability.

The migration and invasion of esophageal squamous cell carcinoma are associated with clinical outcomes, however, the mechanisms remain poorly understood. Here, we found that miR-21 is significantly overexpressed in ESCC, lung cancer, and bladder cancer compared with the adjacent normal tissue. MiR-21 and TPM1 expressions were analyzed by RT-qPCR and WB in 30 ESCC, 10 lung cancer, and 10 bladder cancer clinical specimens, each with matched adjacent normal tissue. Knockdown and overexpression of miR-21 as well as knockdown of TPM1 in ESCC cell lines were performed using synthetic oligonucleotides. TPM1 3′UTR luciferase reporter constructs were used to investigate targeting of TPM1 by miR-21. ESCC migration and invasion were assessed using transwell migration and invasion assays. Inhibition of miR-21 reduced migration and invasion in two ESCC cell lines, and overexpression of miR-21 promoted migration and invasion in vitro. Interestingly, TPM1 exhibited inverse patterns of expression compared with miR-21 in tissues and cell lines. Luciferase reporter assays demonstrated that TPM1 was directly regulated by miR-21. Moreover, the forced overexpression of miR-21 repressed the TPM1 expression, while silencing of miR-21 restored the TPM1 expression in ESCC cell lines. What is more, simultaneous silencing of miR-21 and TPM1 expressions did not alter the migratory and invasive characteristics demonstrating that the effects of miR-21 were mediated through TPM1. In conclusion, the aberrant overexpression of miR-21 is common in cancer and promotes the migration and invasion of ESCC through inhibiting the TPM1 expression. These results suggest that miR-21 may be a novel predictive marker and therapeutic target for treatment of ESCC.

Summary: Despite decades of studies suggesting that the in vivo adipocyte progenitor resides within the vascular niche, the exact nature of this progenitor remains controversial because distinct studies have attributed adipogenic properties to multiple vascular cell types. Using Cre recombinases labeling distinct vascular lineages, we conduct parallel lineage tracing experiments to assess their degree of contribution to de novo adipogenesis. Although we detect occasional adipocytes that were lineage traced by endothelial or mural recombinases, these are rare events. On the other hand, platelet-derived growth factor receptor alpha (PDGFRα)-expressing adventitial or capsular fibroblasts make a significant contribution to adipocytes in all depots and experimental settings tested. Our data also suggest that fibroblasts transition to an intermediate beige adipocyte phenotype prior to differentiating to a mature white adipocyte. These observations, together with histological analyses revealing that adipose tissue fibroblasts express the mural cell marker PDGFRβ, harmonize a highly controversial field with implications for multiple human diseases, including the pandemic of obesity. : Cattaneo et al. used genetic fate mapping in murine models to test the adipogenic potential of distinct cell types of the vascular wall. These parallel lineage-tracing experiments reveal that fibroblasts are the sole vascular cell type with significant adipocyte progenitor activity, giving rise to brown, beige, and white adipocytes. Keywords: adipogenesis, obesity, vascular wall, lineage tracing, endothelium, mural cells, fibroblasts

Summary: To facilitate understanding of human cardiomyocyte (CM) subtype specification, and the study of ventricular CM biology in particular, we developed a broadly applicable strategy for enrichment of ventricular cardiomyocytes (VCMs) derived from human embryonic stem cells (hESCs). A bacterial artificial chromosome transgenic H9 hESC line in which GFP expression was driven by the human ventricular-specific myosin light chain 2 (MYL2) promoter was generated, and screened to identify cell-surface markers specific for MYL2-GFP-expressing VCMs. A CD77+/CD200− cell-surface signature facilitated isolation of >97% cardiac troponin I-positive cells from H9 hESC differentiation cultures, with 65% expressing MYL2-GFP. This study provides a tool for VCM enrichment when using some, but not all, human pluripotent stem cell lines. Tools generated in this study can be utilized toward understanding CM subtype specification, and enriching for VCMs for therapeutic applications. : In this article, Evans and colleagues generated an H9 BAC transgenic reporter cell line and performed a flow cytometry screen to identify a cell-surface signature specific for MYL2-GFP-expressing VCMs. The cell-surface signature, CD77+/CD200−, facilitated isolation of a nearly pure hESC-derived CM population, enriched for VCMs. VCM enrichment was achieved when using some, but not all, human pluripotent stem cell lines. Tools generated in this study serve to advance our understanding of CM subtype specification, commitment, and maturation. Keywords: cardiac differentiation, human embryonic stem cells, ventricular cardiomyocytes, cell-surface marker signature