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Primordial Germ Cell-Like Cells (PGCLCs) are an in vitro model for primordial germ cell development. Here, the authors couple a novel compound reporter with CRISPR screening to identify key genes for exit from pluripotency and acquisition of PGCLC fate; specifically identifying Nr5a2 and Zfp296.

ABSTRACT Globally, dental caries is the most prevalent chronic oral disease and affects roughly half of all children. The aim of this report was to use metagenomic analyses to investigate the relationship between the oral microbiome and caries in preschool children. A total of 25 preschoolers, aged 3 to 5 years old with severe early childhood caries (ECC), and 19 age-matched, caries-free children as controls were recruited. Saliva samples were collected from the participants and were subjected to metagenomic analyses, whereby the oral microbial communities were investigated. The metagenomic analyses revealed substantial microbiota differences between the two groups, indicating apparent shifts of the oral microbiome present in the ECC group. At the species level, the ECC-enriched microbes included Prevotella amnii, Shuttleworthia satelles, Olsenella uli, and Anaeroglobus geminatus. Interestingly, Actinomyces odontolyticus and Actinomyces graevenitzii exhibited apparent differences at the strain level but not the species level between the ECC and control groups. Functional examination showed that the ECC group displayed extensive alterations in metabolic genes/pathways/modules, including enriched functions in sugar metabolism. Finally, an SVM (support vector machine) classifier comprising seven species was developed and generated a moderately good performance in predicting caries onset (area under the receiver operating characteristic curve [AUC] = 78.33%). Together, these findings indicate that caries is associated with considerable changes in the oral microbiome, some of which can potentially be exploited as therapeutic targets or diagnostic markers. (This study has been registered at ClinicalTrials.gov under registration no. NCT02341352.) IMPORTANCE Dental caries is a highly prevalent oral disease that can lead to severe dental damage and may greatly compromise the quality of life of the affected individuals. Previous studies, including those based on 16S rRNA gene, have revealed that the oral microbiota plays a prominent role in development of the disease. But the approach of those studies was limited in analyzing several key microbiome traits, including species- or strain-level composition and functional profile. Here, we performed metagenomic analyses for a cohort of preschool children with or without caries. Our results showed that caries was associated with extensive microbiota differences at various taxonomic and functional levels. Some caries-associated species had not been previously reported, some of which may have significant clinical implications. A microbiome gene catalogue from children with caries was constructed for the first time. The results demonstrated that caries is associated with alterations of the oral microbiome, including changes in microbial composition and metabolic functional profile.

Background This study was designed to explore the profile and potential influencers of the vaginal microbiome (VMB) among asymptomatic pregnant Chinese women and its possible association with pregnancy outcomes. Methods A prospective study was conducted among pregnant Chinese women receiving regular prenatal care at a hospital in Shanghai, China from March 2017 to March 2018. Vaginal swabs were obtained from 113 asymptomatic pregnant women in mid-pregnancy and sequenced by the V3–V4 region of 16S rRNA on an Ion S5™ XL platform. Demographic characteristics and major pregnancy outcomes were collected through questionnaires and electronic medical records. Results The predominant vaginal community state types (CSTs) were CST I (45.1%) and CST III (31.9%). Participants were divided into a lactobacilli-dominant group (LD, CST I/II/III/I–III/V, n = 100, 88.5%) and a less lactobacilli-dominant group (LLD, CST IV-A/B, n = 13, 11.5%). Women in the LLD group showed an increased alpha diversity [median (interquartile range, IQR): 2.41 (1.67, 2.49) vs. 0.30 (0.17, 0.59), P

Summary: Cartilage pellets generated from ectomesenchymal progeny of human pluripotent stem cells (hPSCs) in vitro eventually show signs of commitment of chondrocytes to hypertrophic differentiation. When transplanted subcutaneously, most of the surviving pellets were fully mineralized by 8 weeks. In contrast, treatment with the adenylyl cyclase activator, forskolin, in vitro resulted in slightly enlarged cartilage pellets containing an increased proportion of proliferating immature chondrocytes that expressed very low levels of hypertrophic/terminally matured chondrocyte-specific genes. Forskolin treatment also enhanced hyaline cartilage formation by reducing type I collagen gene expression and increasing sulfated glycosaminoglycan accumulation in the developed cartilage. Chondrogenic mesoderm from hPSCs and dedifferentiated nasal chondrocytes responded similarly to forskolin. Furthermore, forskolin treatment in vitro increased the frequency at which the cartilage pellets maintained unmineralized chondrocytes after subcutaneous transplantation. Thus, the post-transplantational fate of chondrocytes originating from hPSC-derived chondroprogenitors can be controlled during their genesis in vitro. : In this article, Naoki Nakayama and colleagues show that suppression of hypertrophic differentiation and terminal maturation of chondrocytes by forskolin/cAMP treatment during in vitro chondrogenesis from human pluripotent stem cell-derived chondroprogenitors leads to the formation of cartilage consisting of an increased proportion of proliferating, immature chondrocytes with significantly improved ability to be maintained in an unmineralized state in vivo. Keywords: pluripotent stem cell, neural crest, paraxial mesoderm, chondrocyte, hypertrophy, cAMP, forskolin, cartilage, mineralization, endochondral ossification

Abstract Background DNA methylation is a heritable epigenetic mark, enabling stable but reversible gene repression. In mammalian cells, DNA methyltransferases (DNMTs) are responsible for modifying cytosine to 5-methylcytosine (5mC), which can be further oxidized by the TET dioxygenases to ultimately cause DNA demethylation. However, the genome-wide cooperation and functions of these two families of proteins, especially at large under-methylated regions, called canyons, remain largely unknown. Results Here we demonstrate that DNMT3A and TET1 function in a complementary and competitive manner in mouse embryonic stem cells to mediate proper epigenetic landscapes and gene expression. The longer isoform of DNMT3A, DNMT3A1, exhibits significant enrichment at distal promoters and canyon edges, but is excluded from proximal promoters and canyons where TET1 shows prominent binding. Deletion of Tet1 increases DNMT3A1 binding capacity at and around genes with wild-type TET1 binding. However, deletion of Dnmt3a has a minor effect on TET1 binding on chromatin, indicating that TET1 may limit DNA methylation partially by protecting its targets from DNMT3A and establishing boundaries for DNA methylation. Local CpG density may determine their complementary binding patterns and therefore that the methylation landscape is encoded in the DNA sequence. Furthermore, DNMT3A and TET1 impact histone modifications which in turn regulate gene expression. In particular, they regulate Polycomb Repressive Complex 2 (PRC2)-mediated H3K27me3 enrichment to constrain gene expression from bivalent promoters. Conclusions We conclude that DNMT3A and TET1 regulate the epigenome and gene expression at specific targets via their functional interplay.

Summary: Although EZH2 enzymatic inhibitors have shown antitumor effects in EZH2-mutated lymphoma and ARID1A-mutated ovarian cancer, many cancers do not respond because EZH2 can promote cancer independently of its histone methyltransferase activity. Here we identify ZRANB1 as the EZH2 deubiquitinase. ZRANB1 binds, deubiquitinates, and stabilizes EZH2. Depletion of ZRANB1 in breast cancer cells results in EZH2 destabilization and growth inhibition. Systemic delivery of ZRANB1 small interfering RNA (siRNA) leads to marked antitumor and antimetastatic effects in preclinical models of triple-negative breast cancer (TNBC). Intriguingly, a small-molecule inhibitor of ZRANB1 destabilizes EZH2 and inhibits the viability of TNBC cells. In patients with breast cancer, ZRANB1 levels correlate with EZH2 levels and poor survival. These findings suggest the therapeutic potential for targeting the EZH2 deubiquitinase ZRANB1. : Many cancer cells are sensitive to depletion of EZH2 but resistant to EZH2 inhibitors, due to EZH2’s enzyme-independent cancer-promoting function. Zhang et al. identify ZRANB1 as an EZH2 deubiquitinase and a potential anticancer target. Keywords: ZRANB1, EZH2, deubiquitinase, breast cancer

Abstract Background Anopheles sinensis is an important malaria vector in Southeast Asia. The widespread emergence of insecticide resistance in this mosquito species poses a serious threat to the efficacy of malaria control measures, particularly in China. Recently, the whole-genome sequencing and de novo assembly of An. sinensis (China strain) has been finished. A series of insecticide-resistant studies in An. sinensis have also been reported. There is a growing need to integrate these valuable data to provide a comprehensive database for further studies on insecticide-resistant management of An. sinensis. Results A bioinformatics database named An. sinensis genome database (ASGDB) was built. In addition to being a searchable database of published An. sinensis genome sequences and annotation, ASGDB provides in-depth analytical platforms for further understanding of the genomic and genetic data, including visualization of genomic data, orthologous relationship analysis, GO analysis, pathway analysis, expression analysis and resistance-related gene analysis. Moreover, ASGDB provides a panoramic view of insecticide resistance studies in An. sinensis in China. In total, 551 insecticide-resistant phenotypic and genotypic reports on An. sinensis distributed in Chinese malaria-endemic areas since the mid-1980s have been collected, manually edited in the same format and integrated into OpenLayers map-based interface, which allows the international community to assess and exploit the high volume of scattered data much easier. The database has been given the URL: http://www.asgdb.org /. Conclusions ASGDB was built to help users mine data from the genome sequence of An. sinensis easily and effectively, especially with its advantages in insecticide resistance surveillance and control.

Abstract Background Chemical insecticides have hugely reduced the prevalence of vector-borne diseases around the world, but resistance threatens their continued effectiveness. Despite its importance, cuticle resistance is an under-studied area, and exploring the detailed molecular basis of resistance is critical for implementing suitable resistance management strategies. Methods We performed western blotting of cuticular protein CPLCG5 in deltamethrin-susceptible (DS) and laboratory-produced deltamethrin-resistant (DR) strains of Culex pipiens pallens. Immunofluorescence assays using a polyclonal antibody to locate cuticular CPLCG5 in mosquitoes. EM immunohistochemical analysis of the femur segment was used to compare the cuticle in control and CPLCG5-deficient siRNA experimental groups. Results The gene CPLCG5 encodes a cuticle protein that plays an important role in pyrethroid resistance. Based on a prior study, we found that expression of CPLCG5 was higher in the resistant (DR) strain than the susceptible (DS) strain. CPLCG5 transcripts were abundant in white pupae and 1-day-old adults, but expression was dramatically decreased in 3-day-old adults, then remained stable thereafter. Western blotting revealed that the CPLCG5 protein was ~2.2-fold higher in the legs of the DR strain than the DS strain. Immunofluorescence assays revealed CPLCG5 expression in the head, thorax, abdomen, wing, and leg, and expression most abundant in the leg and wing. EM immunohistochemical analysis suggested that the exocuticle thickness of the femur was significantly thinner in the CPLCG5-deficient siCPLCG5 strain (0.717 ± 0.110 μm) than the siNC strain (0.946 ± 0.126 μm). Depletion of CPLCG5 by RNA interference resulted in unorganised laminae and a thinner cuticle. Conclusions The results suggest CPLCG5 participates in pyrethroid resistance by forming a rigid matrix and increasing the thickness of the cuticle.

The grain orientation of Sn-based solder joints on copper pillars under the combined action of electron wind force and temperature gradient greatly affects their electromigration damage. The copper pillars with Sn-1.8Ag lead-free solder on the top was subjected to a current density of 1.5 × 104 A/cm2 at 125 °C to study the electromigration behaviors. The grain orientation was characterized by scanning electron microscopy (SEM) equipped with electron backscattered diffraction (EBSD) detector. Metal dissolution and voids formation in the cathode as well as massive intermetallic compounds(IMC) accumulation in the anode were observed after electromigration. Closer examination of solder joints revealed that the Sn grain whose c-axis perpendicular to electric current may have retarded Cu diffusion to anode and IMC accumulation. In addition, the newly formed Cu6Sn5 exhibited preferred orientation related to the electric current direction.

For linear accelerometers, calibration with a precision centrifuge is a key technology, and the input acceleration imposed on the accelerometer should be accurately obtained in the calibration. However, there are often errors in the installation of sample that make the calibration inaccurate. To solve installation errors and obtain the input acceleration in the calibration of the accelerometer, a calibration method based on the rotation principle using a double turntable centrifuge is proposed in this work. The key operation is that the sub-turntable is rotated to make the input axis of the accelerometer perpendicular to the direction of the centripetal acceleration vector. Models of installation errors of angle and radius were built. Based on these models, the static radius and input acceleration can be obtained accurately, and the calibration of the scale factor, nonlinearity and asymmetry can be implemented. Using this method, measurements of the MEMS accelerometer with a range of ±30 g were carried out. The results show that the discrepancy of performance obtained from different installation positions was smaller than 100 ppm after calibrating the input acceleration. Moreover, the results using this method were consistent with those using the back-calculation method. These results demonstrate that the effectiveness of our proposed method was confirmed. This method can measure the static radius directly eliminating the installation errors of angle and radius, and it simplifies the accelerometer calibration procedure.

The characteristics of shale micro-pore development and its main influencing factors have important theoretical guiding significance for shale gas exploration and resource evaluation. In order to clarify the micro-pore development characteristics of lower Cambrian shale and the main controlling factors of micro-pore development, we used the lower Cambrian Niutitang formation shale, in the Wenshuicun section of the Guizhou Province in southwest China. The micro-pore development characteristics of the shale in the region were studied by argon ion profile field emission scanning electron microscopy and a low-temperature liquid nitrogen adsorption and desorption experimental system. The relationship between micro-pore and kerogen maceral composition, total organic carbon (TOC) content and different mineral content was analyzed in combination with mineral and geochemical characteristics. Inorganic pores (clay mineral pores, dissolution pores and pyrite intergranular pores) and micro-fractures (clay mineral shrinkage crack, tectonic fractures and overpressure fractures) were the main type of pore developed in the shale of the Niutitang formation in the Wenshuicun section, and no organic pores had developed. The pore size of shale is usually 2–50 nm, accounting for 58.33% of shale pores, e.g. mesopores. Clay mineral content has an obvious positive correlation with macropore volume and average pore diameter, and an obvious negative correlation with micropore volume. In addition, the content of feldspar in brittle minerals has a strong negative correlation with macropore volume and average pore diameter, and a strong positive correlation with micropore volume and BET-specific surface area. TOC content and the content of different kerogen macerals have no obvious correlation with the development of shale micropores in this region. It is concluded that inorganic mineral composition is the main controlling factor of micro-pore development within lower Cambrian shale, and organic matter abundance and maceral content have little influence on the micro-pore development. This study provides a case study for the characteristics of micropores in lower Cambrian shale in China.

Our present study investigated the anti-diabetic activity and potential mechanism of β-ecdysterone (β-EC) derived from Cyanotis arachnoidea. In vitro, β-EC exhibited a promising effect on increasing GLUT4 translocation by 1.6 folds and glucose uptake by 1.75 folds in L6 cells. In vivo, KK-Ay mice’s body weight, blood glucose levels and other related blood–lipid indexes can be significantly reduced with β-EC treatment. For the mechanism study, we have found that β-EC exhibited a significant protective effect on insulin resistance (IR) in L6 and HepG2 cells through increasing expressions of IRβ, p-Akt, p-IRS-1 and increasing the expressions of GLUT4 and GLUT2. The phosphorylation of Akt, IRS-1 and the expression of IRβ, GLUT4 and GLUT2 in the liver and skeletal muscle in KK-Ay mice were also significantly ameliorated after 4-weeks treatment with β-EC. According to our present findings, we could conclude that β-EC possessed the potential anti-diabetic effects through activating IRS-1/AKT/GLUT4 and IRS-1/AKT/GLUT2 pathways.

Background/Aims: Evidence indicates a role for dyslipidemia in the development of chronic kidney disease (CKD). However, the association of lipid abnormalities and their ratios with kidney disease using the new CKD Epidemiology Collaboration (CKD-EPI) equation is not well understood. Methods: This cross-sectional study included 48,054 adult subjects. CKD was defined as an estimated glomerular filtration rate

Lead-free perovskites have received remarkable attention because of their nontoxicity, low-cost fabrication, and spectacular properties including controlled bandgap, long diffusion length of charge carrier, large absorption coefficient, and high photoluminescence quantum yield. Compared with the widely investigated polycrystals, single crystals have advantages of lower trap densities, longer diffusion length of carrier, and extended absorption spectrum due to the lack of grain boundaries, which facilitates their potential in different fields including photodetectors, solar cells, X-ray detectors, light-emitting diodes, and so on. Therefore, numerous research focusing on the novel properties, preparation methods, and remarkable progress in applications of lead-free perovskite single crystals (LFPSCs) has been extensively studied. In this review, the current advancements of LFPSCs are briefly summarized, including the synthesis approaches, compositional and interfacial engineering, and stability of several representative systems of LFPSCs as well as the reported practical applications. Finally, the critical challenges which limit the performance of LFPSCs, and their inspiring prospects for further developments are also discussed.

The metal tolerance protein (MTP) family is a very old family with evolutionary conservation and less specific amplification. It seems to retain the original functions of the ancestral genes and plays an important role in maintaining metal homeostasis in plant cells. We identified the potato MTP family members for the first time, the specific and conservative StMPTs were discovered by using systematic and comparative omics. To be surprised, members of the StMTP family seem to have mutated before the evolution of dicotyledon and monocotyledon, and even the loss of the entire subfamily (subfamily G6, G7). Interestingly, StMTP9 represents the conserved structure of the entire subfamily involved in toxic metal regulation. However, the gene structure and transmembrane domain of StMTP8 have undergone specific evolution, showing that the transmembrane domain (Motif13) located at the NH2 terminal has been replaced by the signal peptide domain, so it was selected as the control gene of StMTP9. Through real-time fluorescence quantitative analysis of StMTPs under Cd and Zn stress, a co-expression network was constructed, and it was found that StMTP9 responded significantly to Cd stress, while StMTP8 did the opposite. What excites us is that by introducing StMTPs 8/9 into the ∆ycf1 yeast cadmium-sensitive mutant strain, the functional complementation experiment proved that StMTPs 8/9 can restore Cd tolerance. In particular, StMTP9 can greatly reduce the cadmium content in yeast cells, while StMTP8 cannot. These findings provide a reference for further research on the molecular mechanism of potato toxic metal accumulation.

Classification is a fundamental task for airborne laser scanning (ALS) point cloud processing and applications. This task is challenging due to outdoor scenes with high complexity and point clouds with irregular distribution. Many existing methods based on deep learning techniques have drawbacks, such as complex pre/post-processing steps, an expensive sampling cost, and a limited receptive field size. In this paper, we propose a graph attention feature fusion network (GAFFNet) that can achieve a satisfactory classification performance by capturing wider contextual information of the ALS point cloud. Based on the graph attention mechanism, we first design a neighborhood feature fusion unit and an extended neighborhood feature fusion block, which effectively increases the receptive field for each point. On this basis, we further design a neural network based on encoder–decoder architecture to obtain the semantic features of point clouds at different levels, allowing us to achieve a more accurate classification. We evaluate the performance of our method on a publicly available ALS point cloud dataset provided by the International Society for Photogrammetry and Remote Sensing (ISPRS). The experimental results show that our method can effectively distinguish nine types of ground objects. We achieve more satisfactory results on different evaluation metrics when compared with the results obtained via other approaches.

In a MEMS capacitive accelerometer, there is an offset due to mechanical and electrical factors, and the offset would deteriorate the performance of the accelerometer. Reducing the offset from mechanism would benefit the improvement in performance. Yet, the compositions of the offset are complex and mix together, so it is difficult to decompose the offset to provide guidance for the reduction. In this work, a decomposition method of offset in a MEMS capacitive accelerometer was proposed. The compositions of the offset were first analyzed quantitatively, and methods of measuring key parameters were developed. Based on our proposed decomposition method, the experiment of offset decomposition with a closed-loop MEMS capacitive accelerometer was carried out. The results showed that the offset successfully decomposed, and the major source was from the fabricated gap mismatch in the MEMS sensor. This work provides a new way for analyzing the offset in a MEMS capacitive accelerometer, and it is helpful for purposefully taking steps to reduce the offset and improve accelerometer performance.

Neuroblastoma cell differentiation is regulated by Id proteins. Here, the authors show that CaMKII-mediated phosphorylation of Beclin 1 can activate K63-linked ubiquitination and autophagic degradation of Id proteins uncovering a role for autophagy in cell differentiation.

Aldosterone, with pro-oxidation and pro-autophagy capabilities, plays a key role in liver fibrosis. However, the mechanisms underlying aldosterone-promoted liver sinusoidal endothelial cells (LSECs) defenestration remain unknown. Caveolin 1 (Cav1) displays close links with autophagy and fenestration. Hence, we aim to investigate the role of Cav1-related autophagy in LSECs defenestration. We found the increase of aldosterone/MR (mineralocorticoid receptor) level, oxidation, autophagy, and defenestration in LSECs in the human fibrotic liver, BDL or hyperaldosteronism models; while antagonizing aldosterone or inhibiting autophagy relieved LSECs defenestration in BDL-induced fibrosis or hyperaldosteronism models. In vitro, fenestrae of primary LSECs gradually shrank, along with the down-regulation of the NO-dependent pathway and the augment of the AMPK-dependent autophagy; these effects were aggravated by rapamycin (an autophagy activator) or aldosterone treatment. Additionally, aldosterone increased oxidation mediated by Cav1, reduced ATP generation, and subsequently induced the AMPK-dependent autophagy, leading to the down-regulation of the NO-dependent pathway and LSECs defenestration. These effects were reversed by MR antagonist spironolactone, antioxidants or autophagy inhibitors. Besides, aldosterone enhanced the co-immunoprecipitation of Cav1 with p62 and ubiquitin, and induced Cav1 co-immunofluorescence staining with LC3, ubiquitin, and F-actin in the perinuclear area of LSECs. Furthermore, aldosterone treatment increased the membrane protein level of Cav1, whereas decrease the cytoplasmic protein level of Cav1, indicating that aldosterone induced Cav1-related selective autophagy and F-actin remodeling to promote defenestration. Consequently, Cav1-related selective autophagy initiated by aldosterone-induced oxidation promotes LSECs defenestration via activating the AMPK-ULK1 pathway and inhibiting the NO-dependent pathway.

Abstract Background The assessment and characterization of the gut microbiome has become a focus of research in the area of human autoimmune diseases. Ankylosing spondylitis is an inflammatory autoimmune disease and evidence showed that ankylosing spondylitis may be a microbiome-driven disease. Results To investigate the relationship between the gut microbiome and ankylosing spondylitis, a quantitative metagenomics study based on deep shotgun sequencing was performed, using gut microbial DNA from 211 Chinese individuals. A total of 23,709 genes and 12 metagenomic species were shown to be differentially abundant between ankylosing spondylitis patients and healthy controls. Patients were characterized by a form of gut microbial dysbiosis that is more prominent than previously reported cases with inflammatory bowel disease. Specifically, the ankylosing spondylitis patients demonstrated increases in the abundance of Prevotella melaninogenica, Prevotella copri, and Prevotella sp. C561 and decreases in Bacteroides spp. It is noteworthy that the Bifidobacterium genus, which is commonly used in probiotics, accumulated in the ankylosing spondylitis patients. Diagnostic algorithms were established using a subset of these gut microbial biomarkers. Conclusions Alterations of the gut microbiome are associated with development of ankylosing spondylitis. Our data suggest biomarkers identified in this study might participate in the pathogenesis or development process of ankylosing spondylitis, providing new leads for the development of new diagnostic tools and potential treatments.

Abstract Background The biomass yield of Chlorella PY-ZU1 drastically increased when cultivated under high CO2 condition compared with that cultivated under air condition. However, less attention has been given to the microalgae photosynthetic mechanisms response to different CO2 concentrations. The genetic reasons for the higher growth rate, CO2 fixation rate, and photosynthetic efficiency of microalgal cells under higher CO2 concentration have not been clearly defined yet. Results In this study, the Illumina sequencing and de novo transcriptome assembly of Chlorella PY-ZU1 cells cultivated under 15% CO2 were performed and compared with those of cells grown under air. It was found that carbonic anhydrase (CAs, enzyme for interconversion of bicarbonate to CO2) dramatically decreased to near 0 in 15% CO2-grown cells, which indicated that CO2 molecules directly permeated into cells under high CO2 stress without CO2-concentrating mechanism. Extrapolating from the growth conditions and quantitative Real-Time PCR of CCM-related genes, the K m (CO2) (the minimum intracellular CO2 concentration that rubisco required) of Chlorella PY-ZU1 might be in the range of 80–192 μM. More adenosine triphosphates was saved for carbon fixation-related pathways. The transcript abundance of rubisco (the most important enzyme of CO2 fixation reaction) was 16.3 times higher in 15% CO2-grown cells than that under air. Besides, the transcript abundances of most key genes involved in carbon fixation pathways were also enhanced in 15% CO2-grown cells. Conclusions Carbon fixation and nitrogen metabolism are the two most important metabolisms in the photosynthetic cells. These genes related to the two most metabolisms with significantly differential expressions were beneficial for microalgal growth (2.85 g L−1) under 15% CO2 concentration. Considering the micro and macro growth phenomena of Chlorella PY-ZU1 under different concentrations of CO2 (0.04–60%), CO2 transport pathways responses to different CO2 (0.04–60%) concentrations was reconstructed.

Abstract Background 6-Chloro-2-methoxy-N-(phenylmethyl)-9-acridinamine (BA), a novel sponge-derived compound, has been reported to elicit a cytotoxic effect by inhibiting cell proliferation. Methods In this study, we investigated the anti-tumor effect of BA in human hepatocellular carcinoma (HCC) in vitro and in vivo using SMMC-7721 cells. The impact of BA on SMMC-7721 cells was determined by proliferation (clonogenicity and MTT), apoptosis (flow cytometry with annexin V-FITC labeling) and tumor cell migration (Transwell). Apoptosis-related molecules in the PI3K/AKT signaling pathway were examined via Western blotting. We also evaluated the effects of BA on tumor growth using a xenograft nude mouse model. Results The data showed that BA induced dose-dependent cytotoxicity, anti-proliferation, anti-migration and apoptosis in SMMC-7721 cells, accompanied by activation of caspase-3 and a decreased level of caspase-9. Moreover, BA decreased PI3K and p-AKT levels, which indicated the cytotoxicity of BA through the PI3K/Akt pathway. Finally, we confirmed that BA inhibited tumor growth in an HCC xenograft mouse model. Conclusions We concluded that BA induced apoptosis and decreased PI3K and p-AKT expression in human HCC with no effect on the liver, kidney, spleen or lungs. These findings suggest that BA could provide a novel strategy for the treatment of HCC.

Abstract Background Guangdong Province in the Pearl River Delta of Southeast China is among the areas in the country with the highest rates of avian flu cases. In order to control the outbreak of human-infected H7N9 cases, Guangdong launched a new policy on the central slaughtering of live poultry in 2015. This study aims to evaluate attitudes of consumers and live-poultry workers toward the policy. The live-poultry workers consisted of two sub-groups: live-poultry traders and poultry farm workers. Methods Consumers and live-poultry workers from Guangdong were enrolled by stratified multi-stage random sampling. Online and field surveys were conducted to investigate participants’ attitudes on policy implementation. Questionnaires were developed to quantify participant demographics, to collect information about attitudes toward the policy, and to identify influential factors of policy acceptability. Proportional odds logistics regression was used in the univariate and multivariate analyses. A total of 1449 consumers, 181 live-poultry traders, and 114 poultry farm workers completed the study. Results Policy acceptability percentages among consumers, live-poultry traders, and poultry farm workers were 57.1, 37.9, and 62.6%, respectively. Logistics regression shows that consumers tended not to support the policy if they were males, if they were concerned with the food safety of chilled products, and if they preferred purchasing live poultry. Live-poultry traders tended not to support if they were subsidized by the government, if they were males, if they experienced a drop in trading volume, and if they were unclear whether avian flu was a preventable disease. Finally, poultry farm workers tended not to support if they experienced a drop in trading volume, if they operated a poultry farm on a small to medium scale, and if they experienced inconvenience in their work due to the policy. Conclusions The study reveals a substantial refusal or slowness to accept the policy. Failure to accept the policy results from varying reasons. Among consumers, concern about food safety and dietary preference are two major causes of disapproval. Policy acceptability among live-poultry workers diverges within the two sub-groups. While a large percentage of poultry farm workers accept the policy, the drop in trading and an insufficient subsidy hamper acceptance by live-poultry traders. We recommend that policy-makers promote health education and alleviate the policy impact on trading with a reformed subsidy policy to increase acceptability. These findings are crucial for the prevention of human-infected H7N9 cases in Guangdong.

Abstract Background P73 antisense RNA 1 T (non-protein coding), also known as TP73-AS1, is a long non-coding RNA (lncRNA) which is involved in cell proliferation and the development of tumors. However, the exact effects and molecular mechanisms of TP73-AS1 in hepatocellular carcinoma (HCC) progression are still unknown. The present study is aimed to investigate the detailed functions and the mechanism of TP73-AS1 in regulation of HCC cell proliferation. Methods TP73-AS1 expression in HCC tissues and cell lines was determined using real-time PCR assays; the correlation of TP73-AS1 expression with clinicopathological features of HCC was analyzed. The functions of TP73-AS1 in regulation of HCC cell proliferation was evaluated using MTT and BrdU assays. The candidate upstream miRNAs of HMGB1 were screened using miRcode, miRWalk, miRanda and Target scan, verified using real-time PCR assays. The interaction between TP73-AS1 and miR-200a was confirmed using Luciferase report gene assays. The proten levels of HMGB1 signaling-related factors in response to co-processing TP73-AS1 knockdown and miR-200a inhibition were determined using Western blot assays and ELISA. Further, miR-200a, HMGB1 mRNA and RAGE mRNA and their correlations in HCC tissues were determined. Results TP73-AS1 was upregulated in HCC tissues and cell lines. High TP73-AS1 expression was correlated with worse clinicopathological features, poorer prognosis and shorter survival. Knockdown of TP73-AS1 inhibited the HCC proliferation and the expression levels of HMGB1, RAGE and NF-κB in HCC cells. By using online tools, we screened out several candidate upstream miRNAs of HMGB1, among which miR-200a overexpression inhibited HMGB1 mRNA expression the most significantly. By using luciferase assays, we confirmed that miR-200a could directly bind to TP73-AS1 and the 3’UTR of HMGB1; TP73-AS1 competed with HMGB1 for miR-200a binding. MiR-200a inhibition could up-regulate HMGB1, RAGE, NF-κB expression as well as NF-κB regulated cytokines levels, which could be partially restored by si-TP73-AS1. In HCC tissues, miR-200a was down-regulated while HMGB1 and RAGE were up-regulated; TP73-AS1 was inversely correlated with miR-200a, while positively correlated with HMGB1 and RAGE, respectively. Conclusion Our data indicated that TP73-AS1 might be an oncogenic lncRNA that promoted proliferation of HCC and could be regarded as a therapeutic target in human HCC.

Danggui Buxue Tang (DBT) is an ancient herbal mixture containing Astragali Radix and Angelicae Sinensis Radix, and which are commonly consumed for “qi-invigorating” (i.e., stimulating vital energy/energy metabolism) as traditional Chinese medicine (TCM). The pharmacological activities of DBT in anti-oxidation, estrogenic, hematopoietic, and immunogenic have been reported; however, the role of DBT in cellular energy metabolism has not been determined. Here, we employed an extracellular flux analyzer to evaluate the mitochondrial respiration of cultured H9C2 cardiomyoblasts in present of DBT. The herbal extract of DBT was qualified chemically for the major ingredients, i.e. astragaloside, calycosin, formononetin, Z-ligustilide, and ferulic acid. The anti-oxidant activities of DBT, as well as its major ingredients, were determined by Folin-Ciocalteu assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, and protective effect in tert-butyl hydroperoxide (tBHP)-treated cultured cardiomyoblasts. In addition, a real-time oxygen consumption rate (OCR) in herbal extract-treated cultured cardiomyoblasts was revealed by using a Seahorse extracellular flux analyzer. In addition, the transcript expressions of peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PCG-1α) and other genes relating to mitochondria biogenesis were determined in cardiomyoblasts under different herbal treatments. DBT possessed the strongest anti-oxidant activity and protective effects on the oxidatively stressed cardiomyoblasts. By revealing the OCR in mitochondria, the health state of cultured cardiomyoblasts under DBT was improved via increase of basal respiration, proton leak, non-mitochondria, and adenosine triphosphate (ATP) production. Furthermore, the transcriptional activities of genes responsible for mitochondrial biogenesis and DNA replication were stimulated by application of DBT in cultures.

Objectives: To determine whether pre-existing nephropathy impacts urinary KIM-1 levels, urinary KIM-1 were measured in patients with normal kidney filtration function but either with or without proteinuria. The reference intervals of urinary KIM-1 in adults with normal kidney filtration function but without urine proteinuria were established. Design and methods: 188 urine samples were obtained from adults with normal kidney filtration. 83 of the 188 showed negative urine protein, erythrocytes and leucocytes were used as normal controls. The remaining 105 samples showed at least one abnormal result suggesting possible pre-existing nephropathy. Urinary KIM-1 concentrations were measured using an enzyme-linked immunosorbent assay. Urinary KIM-1 was normalized with urine creatinine concentration. The reference interval for urinary KIM-1 was determined by non-parametric methodology on 147 individuals. Results: The results showed significantly increased urinary KIM-1 concentration in protein positive (protein +, erythrocyte +/−, leucocyte+/-) samples compared to controls (protein-, erythrocyte -, leucocyte -). Urinary KIM-1 concentrations were significantly higher when proteinuria was at trace concentration (0.25 g/L) and correlated with the severity of proteinuria. The creatinine normalized urinary KIM-1 was significantly higher when urine protein was 1 + to 3+ (0.75–5 g/L). The reference interval for urinary KIM-1 was 0.00 (90%CI: 0-0) to 4.19 (90%CI: 3.11–5.62) μg/L, and for creatinine normalized urinary KIM-1 0.00 (90%CI: 0-0) to 0.58 (90%CI: 0.44–0.74) μg/mmol. Conclusions: Baseline urinary KIM-1 concentrations were increased when there was detectable urine protein and correlated with its severity. The urinary KIM-1 concentrations should be interpreted with consideration of urine protein levels in individual patients. Keywords: Urinary KIM-1, Kidney injury, Nephropathy, Reference interval

[This corrects the article DOI: 10.1371/journal.pgen.1005833.].

Objective: Dysbiosis of gastric microbiota such as Helicobacter pylori plays a significant role in pathogenesis and progression of gastric cancer. Our aim was to evaluate the composition and functional effects of gastric microbiota in superficial gastritis (SG) and advanced gastric adenocarcinoma (GC).Methods: We carried out shotgun metagenomic sequencing on gastric wash samples from 6 patients with GC and 5 patients with SG. The taxonomic composition was profiled using MetaPhlAn2 and functional gene pathway was profiled using HUMAnN2. Differences in microbial composition and pathways between the two patient groups were assessed via LEfSe.Results: The gastric microbiota in GC patients was characterized by reduced species richness, enrichment of 13 bacterial taxa and depletion of 31 taxa (q < 0.05). The most representative taxa which were abundant in GC corresponded to the commensals or opportunistic pathogens that usually colonize the oral cavity, including genera Neisseria, Alloprevotella, and Aggregatibacter, species Streptococcus_mitis_oralis_pneumoniae and strain Porphyromonas_endodontalis.t_GCF_000174815. Each of the three GC-associated genera could separate GC from SG completely. In particular, Sphingobium yanoikuyae, a bacterium capable of degrading carcinogenic compounds, was depleted in GC. Functionally, pathways associated with the biosynthesis of lipopolysaccharide (LPS) and L-arginine were enriched in GC, whereas pathways involved in the fermentation of short chain fatty acids (SCFAs) and branched amino acid metabolism were more abundant in SG.Conclusions: Our results present new alterations in the gastric microbiome in patients with GC from a whole-genome perspective, suggesting that microbiome composition and function can be used for prognosis and diagnosis of GC.

The endoplasmic reticulum (ER) Ca2+ sensors stromal interaction molecule 1 (STIM1) and STIM2, which connect ER Ca2+ depletion with extracellular Ca2+ influx, are crucial for the maintenance of Ca2+ homeostasis in mammalian cells. Despite the recent progress in unraveling the role of STIM2 in Ca2+ signaling, the mechanistic underpinnings of its activation remain underexplored. We use an engineering approach to direct ER-resident STIMs to the plasma membrane (PM) while maintaining their correct membrane topology, as well as Förster resonance energy transfer (FRET) sensors that enabled in cellulo real-time monitoring of STIM activities. This allowed us to determine the calcium affinities of STIM1 and STIM2 both in cellulo and in situ, explaining the current discrepancies in the literature. We also identified the key structural determinants, especially the corresponding G residue in STIM1, which define the distinct activation dynamics of STIM2. The chimeric E470G mutation could switch STIM2 from a slow and weak Orai channel activator into a fast and potent one like STIM1 and vice versa. The systemic dissection of STIM2 activation by protein engineering sets the stage for the elucidation of the regulation and function of STIM2-mediated signaling in mammals.

“Yin-Yang” and “Five Elements” theories are the basis theories of Traditional Chinese Medicine (TCM). To probe and clarify the theoretical basis of these ancient Chinese wisdoms, extensive efforts have been taken, however, without a full success. In the classification of TCM herbs, hot, cold and neutral herbs are believed to possess distinct profile of chemical compositions of which the compounds should have different polarity and mass: this view provides a new perspective for further illustration. To understand the chemical properties of TCMs in the classification of “Yin-Yang” and “Five Elements,” 15 commonly used herbs attributed to spleen-meridian were selected for analyses. Chemically standardized water extracts, 50% ethanol extracts and 90% ethanol extracts were prepared and subjected to different analytic measurements. Principle component analysis (PCA) of full spectrum of HPLC, NMR and LC-MS of the extracts were established. The results revealed that the LC-MS profile showed a strong correlation with the “Yin-Yang” classification criterion. The Yang-stimulating herbs generally contain more compounds with lower molecular weight and less polar property. Additionally, a comprehensive anti-oxidative profiles of selected herbs were developed, and the results showed that its correlation with cold and hot properties of TCM, however, was rather low. Taken together, the “Yin-Yang” nature of TCM is closely related to the physical properties of the ingredients, such as polarity and molecular mass; while such classification has little correlation with anti-oxidative property. Therefore, the present results provide a new direction in probing the basic principle of TCM classification.

The in-situ observation of Sn-3.0Ag-0.5Cu solder joints under electromigration was conducted to investigate the microstructure and grain orientation evolution. It was observed that there was a grain rotation phenomenon during current stressing by in-situ electron backscattered diffraction (EBSD). The rotation angle was calculated, which indicated that the grain reorientation led to the decrease of the resistance of solder joints. On the other hand, the orientation of β-Sn played a critical role in determining the migration of Cu atoms in solder joints under current stressing migration. When the angle between the electron flow direction and the c-axis of Sn (defined as α) was close to 0°, massive Cu6Sn5 intermetallic compounds were observed in the solder bulk; however, when α was close to 90°, the migration of the intermetallic compound (IMC) was blocked but many Sn hillocks grew in the anode. Moreover, the low angle boundaries were the fast diffusion channel of Cu atoms while the high grain boundaries in the range of 55°–65° were not favorable to the fast diffusion of Cu atoms.

The latest research has indicated that anti-tumor agents with COX-2 inhibitory activity may benefit their anti-tumor efficiency. A series of sclerotiorin derivatives have been synthesized and screened for their cytotoxic activity against human lung cancer cells A549, breast cancer cells MDA-MB-435 using the MTT method. Among them, compounds 3, 7, 12, 13, 15, 17 showed good cytotoxic activity with IC50 values of 6.39, 9.20, 9.76, 7.75, 9.08, and 8.18 μM, respectively. In addition, all compounds were tested in vitro the COX-2 inhibitory activity. The results disclosed compounds 7, 13, 25 and sclerotiorin showed moderate to good COX-2 inhibition with the inhibitory ratios of 58.7%, 51.1%, 66.1% and 56.1%, respectively. Notably, compound 3 displayed a comparable inhibition ratio (70.6%) to the positive control indomethacin (78.9%). Furthermore, molecular docking was used to rationalize the potential of the sclerotiorin derivatives as COX2 inhibitory agents by predicting their binding energy, binding modes and optimal orientation at the active site of the COX-2. Additionally, the structure-activity relationships (SARS) have been addressed.

Abstract Purpose betatrophin has been reported to boost β cell expansion in insulin resistant states. Pregnancy is a well-recognized physiological state of insulin resistance. Betatrophin levels in pregnant women and their relationships with metabolic variables remain to be elucidated. Methods A total of 49 pregnant women and 31 age-matched unpregnant women with normal glucose regulation (UP-NGR) were included. Among these subjects, according to results from 75 g oral glucose tolerance test (OGTT), 22 women were diagnosed as having gestational diabetes mellitus ( GDM ). Results Our study found that pregnant women, regardless of their glucose regulation status, had remarkably higher triglycerides (TG), total cholesterol (TC), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR) and homeostasis model assessment of β-cell function (HOMA-β). However, GDM patients had much lower HOMA-β compared with those of pregnant women with normal glucose regulation (P-NGR). Participants of the P-NGR group had almost 4 times higher levels of betatrophin than those of the UP-NGR group. Although betatrophin levels were lower in the GDM group than those of the P-NGR group, the difference did not reach statistical significance. Spearman correlation analysis showed that betatrophin levels were positively and significantly associated with total cholesterol, triglycerides, highdensity lipoprotein cholesterol (HDL-c), FINS and HOMA-β. However, adjustments of TC, TG and HDL-c eliminated the association between HOMA-β and betatrophin. Conclusions Pregnant women have significantly higher betatrophin levels in comparison to unpregnant women. Betatrophin levels are positively and significantly associated with β cell function and lipid levels. Furthermore, lipids may contribute to the association between betatrophin and β cell function.

Ascomylactam A was reported for the first time as a new 13-membered-ring macrocyclic alkaloid in 2019 from the mangrove endophytic fungus Didymella sp. CYSK-4 from the South China Sea. The aim of our study was to delineate the effects of ascomylactam A (AsA) on lung cancer cells and explore the antitumor molecular mechanisms underlying of AsA. In vitro, AsA markedly inhibited the cell proliferation with half-maximal inhibitory concentration (IC50) values from 4 to 8 μM on six lung cancer cell lines, respectively. In vivo, AsA suppressed the tumor growth of A549, NCI-H460 and NCI-H1975 xenografts significantly in mice. Furthermore, by analyses of the soft agar colony formation, 5-ethynyl-20-deoxyuridine (EdU) assay, reactive oxygen species (ROS) imaging, flow cytometry and Western blotting, AsA demonstrated the ability to induce cell cycle arrest in G1 and G1/S phases by increasing ROS generation and decreasing of Akt activity. Conversely, ROS inhibitors and overexpression of Akt could decrease cell growth inhibition and cell cycle arrest induced by AsA. Therefore, we believe that AsA blocks the cell cycle via an ROS-dependent Akt/Cyclin D1/Rb signaling pathway, which consequently leads to the observed antitumor effect both in vitro and in vivo. Our results suggest a novel leading compound for antitumor drug development.

Electron beam welding (EBW) is widely used to weld titanium alloy parts such as aero-engine casing and blades. The surface quality after EBW has a significant influence on the aero-engine performance of those parts. We propose a surface treatment method with grinding on a titanium alloy electron beam weld. We analyze the influence of grinding parameters on the characteristics of the grinding surface. The experiment shows the applicability of ground surface by belt grinding on EBW and its impact on aero-engine performance. After belt grinding, both the welded surface and the surface connected with the substrate are smooth. The extra height of the seam was less than 0.2 mm, and the surface roughness (Ra) of the weld after grinding can be less than 0.98 μm. The microstructure of the weld after grinding was analyzed. Two types of bionic shapes were obtained, a sawtooth shape with a width of 40 μm and a height of 10 μm and a wavy shape with a width of 20 μm and a height of 3 μm. From the analysis above, the bionic surface can be obtained by grinding on the weld with an abrasive belt.

In this work, magnetic In _2 S _3 /ZnFe _2 O _4 nanocomposites were fabricated via a two-step solvothermal process, which In _2 S _3 loading amount on ZnFe _2 O _4 support medium could be controlled by changing reaction mass. Investigation revealed that the photocatalytic activity of In _2 S _3 /ZnFe _2 O _4 nanocomposites was uperior to pure In _2 S _3 and ZnFe _2 O _4 for the degradation of MB driven by visible light. The improved photocatalytic activity may be stemming from the synergy effect of In _2 S _3 and ZnFe _2 O _4 , resulting in the enhanced visible light absorbance and higher separation efficiency of the photo-generated charge carriers. Significantly, this sort of nanocomposites could rapidly be separated through an external magnetic field, and five successive cycle experiments have proved that it was stable and recyclable. Finally, a possible mechanism related to the separation of charge and photocatalytic decomposition of MB was also provided.

Due to their unique electromagnetic properties, meta-atom arrays have always been a hotspot to realize all kinds of particular functions, and the research on meta-atom structure has extended from two-dimensions (2D) to three-dimensions (3D) in recent years. With the continuous pursuit of complex 3D meta-atom arrays, the increasing demand for more efficient and more precise nanofabrication methods has encountered challenges. To explore better fabrication methods, we presented a programmable nanofabrication method for a complex 3D meta-atom array based on focused-ion-beam stress-induced deformation (FIB-SID) effect and designed a distinctive nanostructure array composed of periodic 3D meta-atoms to demonstrate the presented method. After successful fabrication of the designed 3D meta-atom arrays, measurements were conducted to investigate the electric/magnetic field properties and infrared spectral characteristics using scanning cathodoluminescence (CL) microscopic imaging and Fourier transform infrared (FTIR) spectroscopy, which revealed a certain excitation mode induced by polarized incident IR light near 8 μm. Besides the programmability for complex 3D meta-atoms and wide applicability of materials, a more significant advantage of the method is that a large-scale array composed of complex 3D meta-atoms can be processed in a quasi-parallel way, which improves the processing efficiency and the consistency of unit cells dramatically.

Abstract The shift of cytokine profile from anti‐ to pro‐inflammatory is the most recognizable sign of labor, although the underlying mechanism remains elusive. Here, we report that the epithelial sodium channel (ENaC) is upregulated and activated in the uterus at labor in mice. Mechanical activation of ENaC results in phosphorylation of CREB and upregulation of pro‐inflammatory cytokines as well as COX‐2/PGE2 in uterine epithelial cells. ENaC expression is also upregulated in mice with RU486‐induced preterm labor as well as in women with preterm labor. Interference with ENaC attenuates mechanically stimulated uterine contractions and significantly delays the RU486‐induced preterm labor in mice. Analysis of a human transcriptome database for maternal–fetus tissue/blood collected at onset of human term and preterm births reveals significant and positive correlation of ENaC with labor‐associated pro‐inflammatory factors in labored birth groups (both term and preterm), but not in non‐labored birth groups. Taken together, the present finding reveals a pro‐inflammatory role of ENaC in labor at term and preterm, suggesting it as a potential target for the prevention and treatment of preterm labor.

Background Burn-out among doctors threatens their own health, and that of their patients. To identify risk factors of the doctor burn-out is vital to improving their health and increasing the quality of healthcare services. This study aims to explore the relationship between work-family conflict (WFC) and burn-out among Chinese doctors and the mediating role of coping styles in this relationship.Methods A cross-sectional survey was conducted in China, with a questionnaire packet which consisted of the Chinese Maslach Burnout Inventory (CMBI), WFC Scale and the Simplified Coping Style Questionnaire (SCSQ). A total of 2530 doctors participated in the survey. Correlation analysis was performed to explore the relationship between CMBI, WFC and SCSQ scores. A linear regression model was set to determine the mediating role of coping styles on the relationship between WFC and burn-out.Results Doctors who had higher scores on work interfering with family conflict, reported more emotional exhaustion (r=0.514, P

The present study was undertaken to investigate the hypoglycemic activity and potential mechanisms of action of a flavonoid-rich extract from Sophora davidii (Franch.) Skeels (SD-FRE) through in vitro and in vivo studies. Four main flavonoids of SD-FRE namely apigenin, maackiain, leachianone A and leachianone B were purified and identified. In vitro, SD-FRE significantly promoted the translocation and expression of glucose transporter 4 (GLUT4) in L6 cells, which was significantly inhibited by Compound C (AMPK inhibitor), but not by Wortmannin (PI3K inhibitor) or Gö6983 (PKC inhibitor). These results indicated that SD-FRE enhanced GLUT4 expression and translocation to the plasma membrane via the AMPK pathway and finally resulted in an increase of glucose uptake. In vivo, using a spontaneously type 2 diabetic model, KK-Ay mice received intragastric administration of SD-FRE for 4 weeks. As a consequence, SD-FRE significantly alleviated the hyperglycemia, glucose intolerance, insulin resistance and hyperlipidemia in these mice. Hepatic steatosis, islet hypertrophy and larger adipocyte size were observed in KK-Ay mice. However, these pathological changes were effectively relieved by SD-FRE treatment. SD-FRE promoted GLUT4 expression and activated AMPK phosphorylation in insulin target tissues (muscle, adipose tissue and liver) of KK-Ay mice, thus facilitating glucose utilization to ameliorate insulin resistance. Regulation of ACC phosphorylation and PPARγ were also involved in the antidiabetic effects of SD-FRE. Taken together, these findings indicated that SD-FRE has the potential to alleviate type 2 diabetes.

Eupatorium adenophorum Spreng. (EA) is a well-known noxious invasive species. Gas chromatography-mass spectrometry (GC-MS) analysis revealed that the essential oil derived from EA (EAEO) is mainly composed of sesquiterpenes. However, the pharmacological value of EAEO in hepatocellular carcinoma (HCC) remains largely unexplored. Herein, we investigated the anti-HCC activities of EAEO, and explored the potential mechanisms of EAEO-induced apoptosis. An MTT assay showed that EAEO inhibited HCC cell proliferation with little toxicity on normal liver cells. Wound healing and FACS assays revealed that EAEO suppressed HCC cell migration and arrested cell cycle, respectively. Moreover, EAEO promoted in vitro HCC cell apoptosis, and EAEO treatment inhibited HepG2 xenografts growth and enhanced apoptotic nucleus of xenografts in HepG2-bearing nude mice. Mechanistically, EAEO significantly decreased the ratio of Bcl-2/Bax and resulted in the activation of caspase-9 and -3. EAEO also reduced the expression of Grp78, which in turn relieved the inhibition of caspase-12 and -7. Meanwhile, EAEO suppressed the phosphorylation of STAT3 and AKT, indicative of its anti-HCC potential. In summary, we determined that EAEO treatment promoted HCC apoptosis via activation of the apoptotic signaling pathway in mitochondria and endoplasmic reticulum, as well as repressed the activity of STAT3 and AKT in HCC cells.

Early mouse development is regulated and accompanied by dynamic changes in chromatin modifications, including G9a-mediated histone H3 lysine 9 dimethylation (H3K9me2). Previously, we provided insights into its role in post-implantation development (Zylicz et al., 2015). Here we explore the impact of depleting the maternally inherited G9a in oocytes on development shortly after fertilisation. We show that G9a accumulates typically at 4 to 8 cell stage to promote timely repression of a subset of 4 cell stage-specific genes. Loss of maternal inheritance of G9a disrupts the gene regulatory network resulting in developmental delay and destabilisation of inner cell mass lineages by the late blastocyst stage. Our results indicate a vital role of this maternally inherited epigenetic regulator in creating conducive conditions for developmental progression and on cell fate choices.

Kurarinone, a flavonoid isolated from Sophora flavescens Aiton, has been reported to have significant antitumor activity. However, the cytotoxic activity of kurarinone against non-small cell lung cancer (NSCLC) cells is still under explored. In our study, we have evaluated the inhibitory effects of kurarinone on the growth of NSCLC both in vivo and in vitro as well as the molecular mechanisms underlying kurarinone-induced A549 cell apoptosis. The results showed that kurarinone effectively inhibited the proliferation of A549 cells with little toxic effects on human bronchial epithelial cell line BEAS-2B. FASC examination and Hoechst 33258 staining assay showed that kurarinone dose-dependently provoked A549 cells apoptosis. Mechanistically, kurarinone significantly decreased the ratio of Bcl-2/Bax, thereby causing the activation of caspase 9 and caspase 3, and reduced the expression of Grp78, which led to relieve the inhibition of caspase-12 and caspase-7, as well as suppressing the activity of AKT. Meanwhile, modeling results from the Surflex-Dock program suggested that residue Ser473 of Akt is a potential binding site for kurarinone. In vivo, kurarinone inhibited the growth of A549 xenograft mouse models without apparent signs of toxicity. Our study indicated that kurarinone has the potential effects of anti-NSCLC, implemented through activating mitochondria apoptosis signaling pathway, as well as repressing the activity of endoplasmic reticulum pathway and AKT in A549 cells.

We propose a data-driven network-based approach to understand the interactions among technologies, products, and customers. Specifically, the approach enables both a qualitative understanding and a quantitative assessment of the impact of technological changes on customers’ co-consideration behaviors (decision of cross-shopping) and as a consequence the product competitions. The uniqueness of the proposed approach is its capability of predicting complex co-consideration relations of products as a network where both descriptive analyses (e.g., network statistics and joint correspondence analysis) and predictive models (e.g., multiple regressions quadratic assignment procedure) are employed. The integrated network analysis approach features three advantages: (1) It provides an effective visual representation of the underlying market structures; (2) It facilitates the evaluation of the correlation between customers’ consideration preferences and product attributes as well as customer demographics; (3) It enables the prediction of market competitions in response to potential technological changes. This paper demonstrates the proposed network-based approach in a vehicle design context. We investigate the impacts of the fuel economy-boosting technologies and the turbocharged engine technology on individual automakers as well as the entire auto industry. The case study provides vehicle engineers with insights into the change of market competitions brought by technological developments and thereby supports attribute decision-making in vehicle design.

Understanding customer preferences in consideration decisions is critical to choice modeling in engineering design. While existing literature has shown that the exogenous effects (e.g., product and customer attributes) are deciding factors in customers’ consideration decisions, it is not clear how the endogenous effects (e.g., the intercompetition among products) would influence such decisions. This paper presents a network-based approach based on Exponential Random Graph Models to study customers’ consideration behaviors according to engineering design. Our proposed approach is capable of modeling the endogenous effects among products through various network structures (e.g., stars and triangles) besides the exogenous effects and predicting whether two products would be conisdered together. To assess the proposed model, we compare it against the dyadic network model that only considers exogenous effects. Using buyer survey data from the China automarket in 2013 and 2014, we evaluate the goodness of fit and the predictive power of the two models. The results show that our model has a better fit and predictive accuracy than the dyadic network model. This underscores the importance of the endogenous effects on customers’ consideration decisions. The insights gained from this research help explain how endogenous effects interact with exogeous effects in affecting customers’ decision-making.

A new prenylated indole diketopiperazine alkaloid, cristatumin F (1), and four known metabolites, echinulin (2), dehydroechinulin (3), neoechinulin A (4) and variecolorin O (5), were isolated from the crude extract of the fungus Eurotium cristatum. The structure of 1 was elucidated primarily by NMR and MS methods. The absolute configuration of 1 was assigned using Marfey’s method applied to its acid hydrolyzate. Cristatumin F (1) showed modest radical scavenging activity against DPPH radicals, and exhibited marginal attenuation of 3T3L1 pre-adipocytes.

Blisk is an important component for the aero-engine, whose surface quality plays a significant role in its performance. So it is necessary to develop a novel and viable method to improve the blisk leading and trailing edge profile shape precision and surface quality. In order to achieve these aims, the method of adaptive belt precision grinding is used to the precision manufacture for weak rigidity deformation of blisk leading and trailing edge in this article. First, the methodology of adaptive belt precision grinding for blade deformation and material removal precision is analyzed by the characteristics of blisk leading and trailing edge, the blade deformation errors are compensated by the detection and control of the contact pressure, and the precision grinding is achieved by controlling rotating speed of the contact wheel according to the contact between the belt and blisk leading and trailing edge. And then, the experiment of the belt grinding technology for the blisk leading and trailing edge is applied to verify this method, and at the same time, the device for adaptive belt precision grinding is illustrated. The experimental results show that for the leading and trailing edge after adaptive belt precision grinding, the surface roughness is less than 0.4 μm, the profile shape errors are less than 20%, and the surface glossy transition between the blade surface and the leading and trailing edge. It is proved that the method of adaptive belt precision grinding might be considered as a viable process for blisk precision manufacture.

As a methodology of resource location and allocation, reverse auctions are one of the most important activities in supply chains. There are four main auction mechanisms in auction theory, ascending-bid auctions, descending-bid auctions, first-price and sealed-bid auctions, and second-price and sealed-bid auctions. Recently, procurement bidding auctions have been widely studied in the aspects of bidding strategy, auction mechanism with different characteristics, behavior and psychology, collusion and its detection method, and the risk management. However, studies addressing the issue of which is the better reverse auction mechanism in cost-down performance are rarely documented. In this work, simulations are performed to study the integrated process of a dynamic online reverse auction and a static sealed-bid reverse auction using timed colored Petri nets based on the contribution of event management and workflow in Petri net theory. In the Petri net models, colored tokens represent bidders’ related data instantly, and transition nodes are in charge of executing bidding process rules whenever they are enabled. In addition, three programming methods including bidder’s bid and auctioneer’s winner set decision-making methods are embedded in the bidding process rules. Then, a hierarchical Petri net model is employed to compare cost-down range performance of dynamic online reverse auction and a static sealed-bid reverse auction, respectively. By modeling a comparison rule through a transition node, a reverse auction mechanism with better cost-down performance can be revealed based on the convergent simulation results.

As modern electronics are developed towards miniaturisation, high-degree integration and intelligentisation, a large amount of heat will be generated during the operation of devices. How to efficiently remove needless heat is becoming more and more crucial for the lifetime and performance of electronic devices. Many efforts have been made to improve the thermal conductivity of polymer composites, which is an important component of electronics. Herein, the authors report on preparation of boron nitride micosphere/epoxy composites. The cross-plane thermal conductivity of the resultant composites is up to 1.03 Wm^‒1K^‒1. This is attributed to the thermally conductive network formed by the peeled hexagonal boron nitride flakes. Thanks to the superior thermal stability of boron nitride micosphere, the boron nitride micosphere/epoxy composite shows a decreased coefficient of thermal expansion (53.47 ppm/K) and an increased glass transition temperature (147.2°C) compared with the pure epoxy resin. In addition, the boron nitride micosphere/epoxy composite exhibits a lower dielectric constant compared with that of the hexagonal boron nitride/epoxy composite. This strategy can potentially pave the way for the design and fabrication of materials with high cross-plane thermal conductivity and lower dielectric properties.

CRISPR-Cas9 based knockout strategies are increasingly used to analyze gene function. However, redundancies and overlapping functions in biological signaling pathways can call for generating multi-gene knockout cells, which remains a relatively laborious process. Here we detail the application of multi-color LentiCRISPR vectors to simultaneously generate single and multiple knockouts in human cells. We provide a complete protocol, including guide RNA design, LentiCRISPR cloning, viral production and transduction, as well as strategies for sorting and screening knockout cells. The validity of the process is demonstrated by the simultaneous deletion of up to four programmed cell death mediators in leukemic cell lines and patient-derived acute lymphoblastic leukemia xenografts, in which single cell cloning is not feasible. This protocol enables any lab with access to basic cellular biology equipment, a biosafety level 2 facility and fluorescence-activated cell sorting capabilities to generate single and multi-gene knockout cell lines or primary cells efficiently within one month.