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Abstract The chloroplast (cp) genome sequence of Mussaenda pubescens, a promising resource that is used as a traditional medicine and drink, is important for understanding the phylogenetic relationships among the Mussaenda family and genetic improvement and reservation. This research represented the first comprehensive description of the morphological characteristics of M. pubescens, as well as an analysis of the complete cp genome and phylogenetic relationship. The results indicated a close relationship between M. pubescens and M. hirsutula based on the morphological characteristics of the flower and leaves. The cp was sequenced using the Illumina NovaSeq 6000 platform. The results indicated the cp genome of M. pubescens spanned a total length of 155,122 bp, including a pair of inverted repeats (IRA and IRB) with a length of 25,871 bp for each region, as well as a large single-copy (LSC) region and a small single-copy (SSC) region with lengths of 85,370 bp and 18,010 bp, respectively. The results of phylogenetic analyses demonstrated that species within the same genus displayed a tendency to group closely together. It was suggested that Antirhea, Cinchona, Mitragyna, Neolamarckia, and Uncaria might have experienced an early divergence. Furthermore, M. hirsutula showed a close genetic connection to M. pubescens, with the two species having partially overlapping distributions in China. This study presents crucial findings regarding the identification, evolution, and phylogenetic research on Mussaenda plants, specifically targeting M. pubescens.

Summary: The incidence of premature myocardial infarction (PMI) has been rising and acute kidney injury (AKI) occurring in PMI patients severely impacts prognosis. This study aimed to develop and validate a prediction model for AKI specific to PMI patients. The MIMIC-Ⅲ-CV and MIMIC-Ⅳ databases were utilized for model derivation of PMI patients. Single-center data served for external validation. There were 571 and 182 AKI patients in the training set (n = 937) and external validation set (n = 292) cohorts, respectively. Finally, a 7-variable model consisting of: Sequential Organ Failure Assessment (SOFA) score, coronary artery bypass grafting (CABG), ICU stay time, loop diuretics, estimated glomerular filtration rate (eGFR) HCO3- and Albumin was developed, achieving an AUC of 0.85 (95% CI: 0.83–0.88) in the training set. External validation also confirmed model robustness. This model may assist clinicians in the early identification of patients at elevated risk for PMI. Further validation is warranted before clinical application.

Eight emerging and six traditional organophosphate esters (OPEs) were investigated in office air from Hangzhou China with all the traditional OPEs and 5 out of 8 emerging OPEs detected. The median concentrations of ∑traditional OPEs and ∑emerging OPEs were 61,200 and 5.81 pg/m3, respectively. Butylphenyl diphenyl phosphate (BPDPP) and trisisopropyltrisphenyl phosphate (TIPPP) were observed for the first time in indoor air and Chinese office, respectively. The levels of ∑traditional OPEs decreased in the following order: summer > autumn > spring > winter. Conversely, no obvious trends were observed for emerging OPEs. ∑traditional OPEs (p 0.05). ∑traditional OPEs, tris(2-chloroethyl) phosphate (TCEP) and TCIPP levels were significantly positively correlated with relative humidity (p

Bitcoin is a cryptocurrency based on blockchain. All historical Bitcoin transactions are stored in the Bitcoin blockchain, but Bitcoin owners are generally unknown. This is the reason for Bitcoin's pseudo-anonymity, therefore it is often used for illegal transactions. Bitcoin addresses are related to Bitcoin users' identities. Some Bitcoin addresses have the potential to be analyzed due to the behavior patterns of Bitcoin transactions. However, existing Bitcoin analysis methods do not consider the fusion of new blocks' data, resulting in low efficiency of Bitcoin address analysis. In order to address this problem, this paper proposes an incremental Bitcoin address cluster method to avoid re-clustering when new block data is added. Besides, a heuristic Bitcoin address clustering algorithm is developed to improve clustering accuracy for the Bitcoin Blockchain. Experimental results show that the proposed method increases Bitcoin address cluster efficiency and accuracy.

Summary: Genome-wide mapping of transcription factors (TFs) is critical to understand their functions. In chromatin immunoprecipitation (ChIP)-seq assay, it’s challenging to study recruitment of low-abundant TFs transiently boud to the genome. Here, we present an optimized protocol using ChIP Next-Gen Seq Sepharose (Staph-seq) to efficiently pull down chromatin complexes. The double size selection promotes sensitive capture of genome-wide protein-DNA associations while eliminating potential Staph A contamination, which is a common problem in protocols using Staph A cells.For complete details on the use and execution of this protocol, please refer to Tao et al. (2020).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Background: Heart failure (HF) patients in intensive care units (ICUs) are rather poorly studied based on varying left ventricular ejection fraction (LVEF) classification. Characteristics and prognosis of patients in ICUs with HF with mildly reduced ejection fraction (HFmrEF), HF with reduced ejection fraction (HFrEF) and HF with preserved ejection fraction (HFpEF) require further clarification. Methods: Data involving clinical information and 4-year follow-up records of HF patients were extracted and integrated from the Medical Information Mart for Intensive Care III (MIMIC-III) database. Tests were carried out to identity differences among these three HF subtypes. Prognostic analyses were performed using Kaplan-Meier survival analysis and Cox proportional-hazards regression modeling. To develop a novel prediction nomogram, forward selection was used as the best-fit model. Prognostic heterogeneity of the subgroups prespecified by stratification factors in pairwise comparisons was presented using forest plots. Results: A total of 4150 patients were enrolled in this study. HFmrEF had the lowest all-cause mortality rate during the 4-year follow-up, which was significantly different from HFrEF and HFpEF (Log-Rank p < 0.001). The Cox proportional-hazards regression model also showed that a comparison of HFrEF versus HFmrEF indicated a hazard ratio (HR) of 0.76 (95% CI 0.61–0.94, p = 0.011) and HFrEF versus HFpEF indicated a HR 0.93 (95% CI 0.82–1.07, p = 0.307). Following a multivariable analysis, 13 factors were confirmed as independent. A new nomogram was established and quantified with a concordance index (C-index) of 0.70 (95% CI 0.67–0.73), and the internal validation indicated the accuracy of the model. Stratification factors such as a history of coronary artery bypass grafting (CABG) and comorbidity of chronic obstructive pulmonary disease (COPD) induced prognostic heterogeneity among the three subtypes. Conclusions: Clinical characteristics and prognosis significantly varied among the three subtypes of HF patients in ICUs, with HFmrEF patients achieving the best prognosis. The novel prediction model, tailored for this population, showed a satisfying prediction ability.

The aim of this paper is to investigate the water pressure effects on hydraulic fracture behavior of concrete with wedge-splitting testing under dynamic loading. Five waterproof strain gauges are stuck along the crack path to observe the fracture process during the experiments. Four silicon water pressure sensors successfully measured the water pressure value on concrete face. The results show that the water pressure on crack faces accelerates the crack propagation of the concrete. The critical values of the splitting force decrease 26.7 and 25.6%, respectively, with the external applied water pressure of 0.2 and 0.4 MPa. Moreover, the hydraulic crack propagation speed increases at the beginning and tends to reach a peak value finally. The peak value of cracking speed is 11.08 m/s, which is high. Under dynamic loading, the water fails to fill the crack and only the trapped water interacts with the crack face. The water pressure is mainly a parabolic curve distribution along the crack path and the peak value decreases with the increase in the crack length.

Objective To investigate the effect of PLGA-PEG scaffold incorporating angiopoietin 1(Ang1) loaded nanoparticle that was injected into the infarcted area of rat heart to protect the heart from infarction and explore potential mechanisms. Methods Rat models of myocardial infarction were replicated,and one week later SD rats were randomly divided into 3 groups:Ang1-PLGA-PEG injection group [injected with 100 μL nanoparticle suspension, about (1.61±0.05)μg Ang1](n=9),Ang1 injection group[injected with PBS solution of angiopoietin1(100 μL, 25 ng/μL)](n=8),the model control group(injected with 100 μL PBS solution)(n=8). Four weeks after injection,cardiac function was examined by echocardiography,scar tissue hyperplasia was measured by Masson staining, cardiomyocyte apoptosis and angiogenesis were detected by immunofluorescence. Results In comparison with other two groups,Ang1-PLGA-PEG injection group showed an increased blood vessel density in the infarcted area and maturity(P＜0.05),which prevented cardiomyocyte apoptosis (P＜0.05) The area of myocardial infarction was significantly decreased(P＜0.05), the remodeling of cardiomyopathy was reduced and these changes improved cardiac function (P＜0.05). Compared with the model control group, pure Ang1 injected into the surrounding area did not show curative effect (P＜0.05). Conclusions Injection of Ang1-PLGA-PEG nanoparticles into the infarcted area of left ventricle may promote vascular proliferation and facilitate the survival of cardiomyocytes,It provides an experimental basis for clinical application.

An increase in bile acid (BA) levels after metabolic surgery is an important mechanism for improving glucose metabolism. However, the mechanisms underlying elevated BA levels and the regulatory mechanism of glucose metabolism remain unclear. In this study, we used the Goto-Kakizaki rat model to investigate the mechanism of BA elevation by comparing side-to-side jejunoileal bypass plus proximal loop ligation (SSJIBL) and bile ileum diversion (BID) as well as to explore the mechanism of BA metabolism in regulating blood glucose. The results showed that the fed blood glucose of rats in both the SSJIBL and BID groups was significantly lower than that of the SHAM group on days 2 and 14 after the operation. The oral glucose tolerance test (OGTT) improved in the SSJIBL and BID groups at day 14 postoperatively. The expression of CYP27A1 in the livers of the SSJIBL and BID groups was significantly increased. In addition, total serum BA levels in the SSJIBL and BID groups were significantly increased. Moreover, serum levels of lithocholic acid (LCA) and deoxycholic acid (DCA) were significantly higher in the SSJIBL group than in the SHAM group and negatively correlated with the area under the glucose tolerance curve (AUC-OGTT). In conclusion, increased BA synthesis may be an important cause of elevated total serum BA levels, and LCA and DCA are closely associated with improved glucose metabolism.

Carbon paper is one of the most commonly used materials for the gas diffusion layer of proton exchange membrane fuel cell, and its performance is the premise to ensure the stable operation of the cell. In order to improve the structure and properties of carbon paper and make it better meet the performance requirements of fuel cell, this study was conducted to press different amounts of graphite plates in the carbonization process to control the applied pressure and analyze the microstructure evolution and mechanism of carbon paper under different pressure. In addition, the effects of applied pressure on the in-plane resistivity, pore size distribution, surface roughness and mechanical properties of carbon paper were also studied. The results show that applying pressure helps to restrain the cracking of resin carbon, reduce the generation of cracks and improve graphitization degree of carbon paper to a certain extent. Furthermore, with the increase of applied pressure, the surface roughness of carbon paper is reduced gradually, while the average and maximum apertures also decrease. When the applied pressure reaches 3600 Pa, compared with the samples without pressure, the in-plane resistivity of carbon paper decreases from 17.15 mΩ·cm to 12.79 mΩ·cm, the tensile strength and flexural strength are improved by 150% and 82.9% respectively.

Abstract Background Fungus infection in staple grains affects the food storage and threatens food security. The Aspergillus flavus is known to infect multiple grains and produce mycotoxin Aflatoxin B1, which is mutagenic, teratogenic and causes immunosuppression in animals. However, the molecular mechanism of maize resistance to A. flavus is largely unknown. Results Here we used corn kernels to investigate resistance genes to A. flavus using genome-wide association study (GWAS) of 313 inbred lines. We characterized the resistance levels of kernels after inoculating with A. flavus. The GWAS with 558,529 SNPs identified four associated loci involving 29 candidate genes that were linked to seed development, resistance or infection, and involved in signal pathways, seed development, germination, dormancy, epigenetic modification, and antimicrobial activity. In addition, a few candidate genes were also associated with several G-protein signaling and phytohormones that might involve in synergistic work conferring different resistance during seed development. Expression of 16 genes out of 29 during kernel development was also associated with resistance levels. Conclusions We characterized the resistance levels of 313 maize kernels after inoculating with A. flavus, and found four associated loci and 16 candidate maize genes. The expressed 16 genes involved in kernel structure and kernel composition most likely contribute to mature maize kernels’ resistance to A. flavus, and in particular, in the development of pericarp. The linked candidate genes could be experimentally transformed to validate and manipulate fungal resistance. Thus this result adds value to maize kernels in breeding programs.

Aspergillus flavus is a fungus that produces aflatoxin B1, one of the most carcinogenic secondary metabolites. Understanding the regulation mechanism of aflatoxin biosynthesis in this fungus requires precise methods for genomic integration of mutant alleles. To avoid the disadvantage of DNA integration into the genome by non-homologous or ectopic recombination, we developed a novel strategy for site-specific integration of foreign DNA by using a carboxin-resistant sdh2R allele (His 249 Leu). Our results demonstrated that the transformants were generated with a high efficiency (>96%) of correct integration into the sdh2-lcus of the genome of A. flavus NRRL 3357. The advantage of this method is that introduction of the eGFP expression cassette into the sdh2-locus had little effect on fungal growth and virulence while also being rapid and efficient. This system will be a valuable tool for genetic manipulation in A. flavus. To the best of our knowledge, this is the first report on the efficient site-specific integration at the sdh2-locus in the genome of Aspergillus.

Abstract Background Micronuclei (MNi) are extensively used to evaluate genotoxic effects and chromosome instability. However, the roles of kinetochore of MN in mitosis have not been completely addressed. Methods The HeLa CENP B-GFP H2B-mCherry cells are applied to address these questions via the long-term live-cell imaging. In the cells, the kinetochore-positive micronucleus (K+MN) contained CENP B-GFP, while the kinetochore-negative micronucleus (K−MN) did not. Results K−MN-bearing cells produced much more chromosome fragments than did MN-free cells. Most of the chromosome fragments eventually merged into K−MNi. K+MN-bearing cells yielded more kinetochore-positive lagging chromosomes (K+LCs) and K+MNi than MN-free cells did. The results suggested the differences in the fates of K+MNi and K−MNi in mitosis. The cycle of K−MN → Chromosome fragment → K−MN may occur in generations of K−MN-bearing cells, while part of K+MNi might reincorporate into the main nucleus. The K+MN-bearing cells prolonged significantly duration of mitosis compared with MN-free cells. The presence of micronuclei, regardless of K−MN and K+MN, enhanced apoptosis cell death. And K+MN-bearing cells were inclined to apoptosis more than K−MN-bearing cells. The results suggested differences in fates between K−MN-bearing and K+MN-bearing cells. Conclusions Kinetochore determined the fates of micronuclei. Kinetochore in micronuclei indirectly prolonged the duration of mitosis. Kinetochore enhanced cytotoxicity of micronuclei. Our data are direct evidences showing the roles of kinetochore of micronucleus in mitosis of HeLa cells.

Radiation-induced loss of the hematopoietic stem cell progenitor population compromises bone marrow regeneration and development of mature blood cells. Failure to rescue bone marrow functions results in fatal consequences from hematopoietic injury, systemic infections, and sepsis. So far, bone marrow transplant is the only effective option, which partially minimizes radiation-induced hematopoietic toxicities. However, a bone marrow transplant will require HLA matching, which will not be feasible in large casualty settings such as a nuclear accident or an act of terrorism. In this study we demonstrated that human peripheral blood mononuclear cell-derived myeloid committed progenitor cells can mitigate radiation-induced bone marrow toxicity and improve survival in mice. These cells can rescue the recipient’s hematopoietic stem cells from radiation toxicity even when administered up to 24 h after radiation exposure and can be subjected to allogenic transplant without GVHD development. Transplanted cells deliver sEVs enriched with regenerative and immune-modulatory paracrine signals to mitigate radiation-induced hematopoietic toxicity. This provides a natural polypharmacy solution against a complex injury process. In summary, myeloid committed progenitor cells can be prepared from blood cells as an off-the-shelf alternative to invasive bone marrow harvesting and can be administered in an allogenic setting to mitigate hematopoietic acute radiation syndrome.

The basic leucine zipper (bZIP) is an important transcription factor required for fungal development, nutrient utilization, biosynthesis of secondary metabolites, and defense against various stresses. Aspergillus flavus is a major producer of aflatoxin and an opportunistic fungus on a wide range of hosts. However, little is known about the role of most bZIP genes in A. flavus. In this study, we developed a high-throughput gene knockout method based on an Agrobacterium-mediated transformation system. Gene knockout construction by yeast recombinational cloning and screening of the null mutants by double fluorescence provides an efficient way to construct gene-deleted mutants for this multinucleate fungus. We deleted 15 bZIP genes in A. flavus. Twelve of these genes were identified and characterized in this strain for the first time. The phenotypic analysis of these mutants showed that the 15 bZIP genes play a diverse role in mycelial growth (eight genes), conidiation (13 genes), aflatoxin biosynthesis (10 genes), oxidative stress response (11 genes), cell wall stress (five genes), osmotic stress (three genes), acid and alkali stress (four genes), and virulence to kernels (nine genes). Impressively, all 15 genes were involved in the development of sclerotia, and the respective deletion mutants of five of them did not produce sclerotia. Moreover, MetR was involved in this biological process. In addition, HapX and MetR play important roles in the adaptation to excessive iron and sulfur metabolism, respectively. These studies provide comprehensive insights into the role of bZIP transcription factors in this aflatoxigenic fungus of global significance.

Abstract Although several regulatory pathways have been reported for Aspergillus flavus, the regulation of aflatoxin production and mycelial growth under different temperatures remains unclear. In this study, A. flavus differentially expressed genes (DEGs) and regulatory pathways were analyzed under three temperatures, by strand‐specific RNA‐Seq. Results show that a total of 2,428 and 1,474 DEGs were identified in fungal mycelia cultured at 20°C and 37°C, respectively, as compared with the control (28°C). Approximately ~ 79% of DEGs in the 37°C samples were up‐regulated genes, while ~ 63% of DEGs in the 20°C samples were down‐regulated genes. Most of the DEG pathways enriched by lower temperatures differed from those enriched by higher temperatures, while only a small portion of the pathways were shared by A. flavus grown under different temperatures. Aflatoxin biosynthesis, Butanoate metabolism, oxidation–reduction process, and benzene‐containing compound metabolic process were the shared down‐regulated pathways, while steroid biosynthesis, oxidoreductase activity, cellular protein modification process, DNA binding, protein complex were the shared up‐regulated pathways between lower and higher temperatures. The shared genes and pathways are the key regulatory candidates for aflatoxin biosynthesis with changes of temperature. In addition, the identification of both up‐regulated and down‐regulated genes provides a useful gene set for further investigation of the aflatoxin biosynthesis among Aspergillus.

Objective: To confirm the association between baseline blood pressure (BP) levels and the methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism in patients with essential hypertension. Methods: A total of 347 patients were enrolled from the Dongzhi community in Anhui Province, China. The C677T polymorphism of the MTHFR gene was detected using high-throughput TaqMan allelic discrimination assay. Baseline BP was measured using a standardized mercury-gravity monometer. Results: In the whole sample, the frequency of the MTHFR C677T genotypes CC, CT, and TT were 38.6%, 48.1%, and 13.3%, respectively. In a recessive model (CC+CT versus TT genotypes), baseline diastolic blood pressure (DBP) was significantly higher in patients with the TT genotype compared to those with the CT or CC genotypes (P= 0.013). We also divided all patients into three groups based on the tertiles of the baseline BP distribution. Compared to subjects in the lowest tertile of DBP, the adjusted odds of having the TT genotype among subjects in the highest tertile was 2.6 (95% CI: 1.1 to 6.2). However, no significant associations were observed between baseline systolic blood pressure (SBP) and the MTHFR C677T polymorphism. Conclusions: The MTHFR gene polymorphism could be an important genetic determinant of baseline DBP levels in Chinese essential hypertensive patients.

This study was a retrospective analysis of the efficacy of thrombectomy plus local catheter-directed thrombolysis (CDT) for the treatment of lower extremity deep venous thrombosis (LDVT).

Abstract A recent algicidal mode indicates that fungal mycelia can wrap and eliminate almost all co-cultivated algal cells within a short time span. However, the underlying molecular mechanism is rarely understood. We applied proteomic analysis to investigate the algicidal process of Trametes versicolor F21a and identified 3,754 fungal proteins. Of these, 30 fungal enzymes with endo- or exoglycosidase activities such as β-1,3-glucanase, α-galactosidase, α-glucosidase, alginate lyase and chondroitin lyase were significantly up-regulated. These proteins belong to Glycoside Hydrolases, Auxiliary Activities, Carbohydrate Esterases and Polysaccharide Lyases, suggesting that these enzymes may degrade lipopolysaccharides, peptidoglycans and alginic acid of algal cells. Additionally, peptidase, exonuclease, manganese peroxidase and cytochrome c peroxidase, which decompose proteins and DNA or convert other small molecules of algal cells, could be other major decomposition enzymes. Gene Ontology and KEGG pathway enrichment analysis demonstrated that pyruvate metabolism and tricarboxylic acid cycle pathways play a critical role in response to adverse environment via increasing energy production to synthesize lytic enzymes or uptake molecules. Carbon metabolism, selenocompound metabolism, sulfur assimilation and metabolism, as well as several amino acid biosynthesis pathways could play vital roles in the synthesis of nutrients required by fungal mycelia.

The efficacy of eleven essential oils (EOs) against Aspergillus flavus NRRL 3357 was investigated. The highest antifungal activity against this aflatoxigenic fungus was exhibited by cinnamon, oregano and lemongrass, which showed low minimum inhibitory concentration (MIC) values under vapor conditions. Interactions of the three EOs were evaluated by the fractional inhibition concentration index (FICI), and the composite essential oils (CEO) showed synergistic inhibitory activities. Chemical analysis of the composite essential oils of cinnamon, oregano, and lemongrass (COL-CEO) revealed that (Z)-citral (33.44%), (E)-citral (32.88%) and carvacrol (19.84%) were the dominant components, followed by limonene (4.29%) and cinnamaldehyde (3.76%). COL-CEO not only inhibited fungal growth but also decreased aflatoxin B1 production by A. flavus. Downregulation of the relative expression of aflatoxin genes in the aflatoxin biosynthetic pathway by COL-CEO revealed its anti-aflatoxigenic mechanism. COL-CEO could also affect the colonization of A. flavus on maize grains. Therefore, COL-CEO may be considered as a potential natural antifungal agent, which could be used for the storage of maize and other grains.