Liver transcriptomic and proteomic analyses provide new insight into the pathogenesis of liver fibrosis in mice.

BACKGROUND: Liver fibrosis (LF) is a kind of progressive liver injury reaction. The goal of this study was to achieve a more detailed understanding of the molecular changes in response to CCl4-induced LF through the identification of a differentially expressed liver transcriptomic and proteomic. RESULTS: A total of 1224 differentially expressed genes (DEGs) and 302 differentially expressed proteins (DEPs) were significantly identified at the transcriptomic and proteomic level, respectively, and 69 genes (hereafter "cor-DEGs-DEPs" genes) were detected at both levels. Pathway enrichment analysis showed that these cor-DEGs-DEPs genes were significantly enriched in 133 pathways. Importantly, among the cor-DEGs-DEPs genes, Gstm1, Gstm3, Ephx1 and Gstp1 were shown to be associated with metabolic pathways, and confirmed by RT-qPCR and parallel reaction monitoring (PRM) verification. CONCLUSIONS: Through the combined analysis of transcriptomic and proteomic data, this study provides valuable insights into the potential mechanism of the pathogenesis of LF, and lays a theoretical foundation for the further development of targeted therapy for LF.

Deep sequencing of the Sanghuangporus vaninii transcriptome reveals dynamic landscapes of candidate genes involved in the biosynthesis of active compounds.

The medicinal fungus Sanghuang produces diverse bioactive compounds and is widely used in Asian countries. However, little is known about the genes and pathways involved in the biosynthesis of these active compounds. Based on our previous study providing Sanghuangporus vaninii genomic information, the transcriptomes of MY (mycelium), OY (1-year-old fruiting bodies), and TY (3-year-old fruiting bodies) were determined in this study. A significant number of genes (4774) were up- or downregulated between mycelium and fruiting bodies, but only 1422 differentially expressed genes were detected between OY and TY. 138 genes encoding P450s were identified in the fungal genome and grouped into 25 P450 families; more than 64% (88) of the genes were significantly differentially expressed between the mycelium and fruiting body, suggesting that these P450s are involved in fungal sexual development. Importantly, the expression of genes involved in bioactive compound (triterpenoids, polysaccharides, and flavonoids) biosynthesis in asexual (cultured with solid and liquid media) and sexual stages was explored and combined with transcriptome and quantitative PCR analyses. More genes involved in the biosynthesis of bioactive compounds were expressed more highly in mycelium than in fruiting bodies under liquid medium culture compared with solid medium culture, which was consistent with the yields of different bioactive compounds, suggesting that liquid fermentation of S. vaninii Kangneng can be used to obtain these bioactive compounds. A comprehensive understanding of the genomic information of S. vaninii will facilitate its potential use in pharmacological and industrial applications.

Genes involved in Beauveria bassiana infection to Galleria mellonella.

The ascomycete fungus Beauveria bassiana is a natural pathogen of hundreds of insect species and is commercially produced as an environmentally friendly mycoinsecticide. Many genes involved in fungal insecticide infection have been identified but few have been further explored. In this study, we constructed three transcriptomes of B. bassiana at 24, 48 and 72 h post infection of insect pests (BbI) or control (BbC). There were 3148, 3613 and 4922 genes differentially expressed at 24, 48 and 72 h post BbI/BbC infection, respectively. A large number of genes and pathways involved in infection were identified. To further analyze those genes, expression patterns across different infection stages (0, 12, 24, 36, 48, 60, 72 and 84 h) were studied using quantitative RT-PCR. This analysis showed that the infection-related genes could be divided into four patterns: highly expressed throughout the whole infection process (thioredoxin 1); highly expressed during early stages of infection but lowly expressed after the insect death (adhesin protein Mad1); lowly expressed during early infection but highly expressed after insect death (cation transporter, OpS13); or lowly expressed across the entire infection process (catalase protein). The data provide novel insights into the insect-pathogen interaction and help to uncover the molecular mechanisms involved in fungal infection of insect pests.

Optimized Ultrasound Conditions for Enhanced Sensitivity of Molecular Beacons in the Detection of MDR1 mRNA in Living Cells.

P-glycoprotein (P-gp), aprognostic indicator for chemotherapy failure, is encoded by multidrug resistance gene (MDR1). MDR1 mRNA expression could serve as a guidance for personalized medicine. However, the traditional PCR process for mRNA measurement is complicated and cannot realize the real-time detection of mRNA in living single cells. In this work, optimized gold nanoparticle-based molecular beacons were employed to determine MDR1 mRNA levels in living cancer cells. To improve detection sensitivity, ultrasound (US) irradiation was applied to facilitate and enhance cellular uptake of hairpin DNA-coated gold nanoparticle (hDAuNP). The US conditions including irradiation power, exposure time, duty cycle, and incubation time were optimized. The slight difference in MDR1 expression manipulated by siRNA silence could be recognized by US assisted hDAuNP beacons; a 10-fold increase of detection sensitivity was achieved compared with the nonultrasound assistance. Meanwhile, the detection cycle could be shortened from 12 to 2 h. Furthermore, this hDAuNP beacon can serve as an antisense agent to down-regulate P-gp expression and to reverse drug resistance of MCF-7/Adr cells to doxorubicin. Our results demonstrated that the MDR1 hDAuNP beacon assisted by US irradiation had great potential to predict chemotherapy sensitivity and to overcome multidrug resistance in cancer cells and was thus a promising tool for individualized medicine.

Joint analysis of the metabolomics and transcriptomics uncovers the dysregulated network and develops the diagnostic model of high-risk neuroblastoma.

High-risk neuroblastoma (HR-NB) has a significantly lower survival rate compared to low- and intermediate-risk NB (LIR-NB) due to the lack of risk classification diagnostic models and effective therapeutic targets. The present study aims to characterize the differences between neuroblastomas with different risks through transcriptomic and metabolomic, and establish an early diagnostic model for risk classification of neuroblastoma.Plasma samples from 58 HR-NB and 38 LIR-NB patients were used for metabolomics analysis. Meanwhile, NB tissue samples from 32 HR-NB and 23 LIR-NB patients were used for transcriptomics analysis. In particular, integrative metabolomics and transcriptomic analysis was performed between HR-NB and LIR-NB. A total of 44 metabolites (P < 0.05 and fold change > 1.5) were altered, including 12 that increased and 32 that decreased in HR-NB. A total of 1,408 mRNAs (P < 0.05 and |log2(fold change)|> 1) showed significantly altered in HR-NB, of which 1,116 were upregulated and 292 were downregulated. Joint analysis of both omic data identified 4 aberrant pathways (P < 0.05 and impact ≥ 0.5) consisting of glycerolipid metabolism, retinol metabolism, arginine biosynthesis and linoleic acid metabolism. Importantly, a HR-NB risk classification diagnostic model was developed using plasma circulating-free S100A9, CDK2, and UNC5D, with an area under receiver operating characteristic curve of 0.837 where the sensitivity and specificity in the validation set were both 80.0%. This study presents a novel pioneering study demonstrating the metabolomics and transcriptomics profiles of HR-NB. The glycerolipid metabolism, retinol metabolism, arginine biosynthesis and linoleic acid metabolism were altered in HR-NB. The risk classification diagnostic model based on S100A9, CDK2, and UNC5D can be clinically used for HR-NB risk classification.

MicroRNA-like RNA Functions Are Required for the Biosynthesis of Active Compounds in the Medicinal Fungus Sanghuangporus vaninii.

miRNA-like RNAs (milRNAs) have been recognized as sequence-specific regulators of posttranscriptional regulation of gene expression in eukaryotes. However, the functions of hundreds of fungal milRNAs in the biosynthesis of metabolic components are obscure. Sanghuangporus produces diverse bioactive compounds and is widely used in Asian countries. Here, genes encoding two Dicers, four Argonautes, and four RdRPs were identified and characterized in Sanghuangporus vanini. Due to the lack of an efficient gene manipulation system, the efficacy of spray-induced gene silencing (SIGS) was determined in S. vanini, which showed efficient double-stranded RNA (dsRNA) uptake and gene silencing efficiency. SIGS-mediated gene knockdown showed that SVRDRP-3, SVRDRP-4, SVDICER-1, and SVDICER-2 were critical for mycelial biomass, flavonoid, triterpenoid, and polysaccharide production. Illumina deep sequencing was performed to characterize the milRNAs from S. vanini mycelium and fruiting body. A total of 31 milRNAs were identified, out of which, SvmilR10, SvmilR17, and SvmilR33 were Svrdrp-4- and Svdicer-1-dependent milRNAs. Importantly, SIGS-mediated overexpression of SvmilR10 and SvmilR33 resulted in significant changes in the yields of flavonoids, triterpenoids, and polysaccharides. Further analysis showed that these milRNA target genes encoding the retrotransposon-derived protein PEG1 and histone-lysine N-methyltransferase were potentially downregulated in the milRNA overexpressing strain. Our results revealed that S. vanini has high external dsRNA and small RNA uptake efficiency and that milRNAs may play crucial regulatory roles in the biosynthesis of bioactive compounds. IMPORTANCE Fungi can take up environmental RNA that can silence fungal genes with RNA interference, which prompts the development of SIGS. Efficient dsRNA and milRNA uptake in S. vanini, successful dsRNA-targeted gene block, and the increase in intracellular miRNA abundance showed that SIGS technology is an effective and powerful tool for the functional dissection of fungal genes and millRNAs. We found that the RdRP, Dicer, and Argonaute genes are critical for mycelial biomass and bioactive compound production. Our study also demonstrated that overexpressed SVRDRP-4- and SVDICER-1-dependent milRNAs (SvmilR10 and SvmilR33) led to significant changes in the yields of the three active compounds. This study not only provides the first report on SIGS-based gene and milRNA function exploration, but also provides a theoretical platform for exploration of the functions of milRNAs involved in biosynthesis of metabolic compounds in fungi.

Impact of Minerals and Sealing Systems on the Pore Characteristics of the Qiongzhusi Formation Shale in the Southern Sichuan Basin.

The characteristics, distribution, and preservation of pores are vital in controlling the storage and distribution of shale gas. The Qiongzhusi Formation shales taken from different members with similar tectonic and thermal evolutions were used to evaluate the response of pore characteristics to minerals and sealing systems using field-emission scanning electron microscopy and gas adsorption. Because of differences in mineral structure and arrangement, feldspar, organic matter (OM)-clay, OM-rutile, and OM-apatite aggregates facilitate multiple types of pores in the shale and influence the relative proportions of surface porosity for different types of pores owing to differences in mineral structure and arrangement. Rigid frameworks and pressure shadows formed by rigid minerals and OM-mineral aggregates preserved OM and pores to some extent. The sealing capacity of the floor controls the sealing system and hydrocarbon expulsion efficiency of the Qiongzhusi Formation in different members. During thermal evolution, the amount of hydrocarbons generated and expelled affected the stress equilibrium state between the pore pressure and external stress, influencing the compaction intensity of shales. The OM pore development characteristics were evolved with variation in the stress equilibrium state in different sealing systems. Once the stress equilibrium state was disrupted, the OM pores deformed, narrowed, or even closed under the influence of compaction owing to the loss of overpressure support. The pore characteristics of the Qiongzhusi Formation shales responded significantly to different sealing systems. A few OM pores are flat and slitlike in the open system, whereas numerous OM pores are round and elliptical in the semiopen system. Meanwhile, the average diameter of the OM pores in the open system was reduced by approximately 40.2% compared with that of the semiopen system. Furthermore, the pore volume and specific surface area of the mesopores for open system shales were reduced by 38.4% and 37.7%, respectively, compared to the semiopen system. These findings will improve the understanding of the distribution and preservation of pore in shale and help assess the sweet-spot members for the Qiongzhusi Formation shale gas.

Immunotranscriptome analysis of Plutella xylostella reveals differences in innate immune responses to low- and high-virulence Beauveria bassiana strain challenges.

BACKGROUND: Entomopathogenic fungi have developed multiple strategies to overcome the immune defenses of their target insects, whereas insect pests have devised various defense mechanisms to combat fungal infection. However, differences in the molecular mechanisms of the innate immune defense strategies of insects upon infection with different fungal strains from the same species have not been reported. RESULTS: Two Beauveria bassiana strains were obtained that significantly varied in their pathogenicity but were comparable in terms of growth, conidial yield, and cuticle penetration. To investigate the molecular mechanisms underlying the immune response of Plutella xylostella infected with these two strains, RNA-Seq was performed 48 h after infection. A total of 1027 differentially expressed genes (DEGs) were identified, and more than 200 DEGs were enriched in Kyoto Encyclopedia of Genes and Genome (KEGG) pathways involved in disease response, revealing differences in the immune response of P. xylostella to different B. bassiana infections at 48 h. Twenty-eight of the DEGs were related to innate immune functions, such as pathogen recognition, immune system activation and antimicrobial reactions. RNA interference (RNAi)-mediated gene silencing assays showed that PxApoLIII and PxCSP played critical roles in the P. xylostella immune response. PxApoLIII was expressed at higher levels during infection with the high-virulence strain, whereas PxCSP showed the opposite expression pattern during infection with the low-virulence strain, indicating that PxApoLIII and PxCSP might participate in P. xylostella innate immune defense against high- and low-virulence B. bassiana strains. CONCLUSION: The present findings demonstrate that strains of a single species of pathogenic fungi that differ in virulence can induce the expression of different genes in P. xylostella. These results advance our knowledge of the molecular mechanisms underlying fungi-pest interactions.

Comprehensive Analysis of the Transcriptome-Wide m6A Methylation Modification Difference in Liver Fibrosis Mice by High-Throughput m6A Sequencing.

N6-Methyladenosine (m6A), a unique and common mRNA modification method in eukaryotes, is involved in the occurrence and development of many diseases. Liver fibrosis (LF) is a common response to chronic liver injury and may lead to cirrhosis and even liver cancer. However, the involvement of m6A methylation in the development of LF is still unknown. In this study, we performed a systematic evaluation of hepatic genome-wide m6A modification and mRNA expression by m6A-seq and RNA-seq using LF mice. There were 3,315 genes with significant differential m6A levels, of which 2,498 were hypermethylated and 817 hypomethylated. GO and KEGG analyses illustrated that differentially expressed m6A genes were closely correlated with processes such as the endoplasmic reticulum stress response, PPAR signaling pathway and TGF-ß signaling pathway. Moreover, a total of 90 genes had both a significant change in the m6A level and mRNA expression shown by joint analysis of m6A-seq and RNA-seq. Hence, the critical elements of m6A modification, including methyltransferase WTAP, demethylases ALKBH5 and binding proteins YTHDF1 were confirmed by RT-qPCR and Western blot. In an additional cell experiment, we also observed that the decreased expression of WTAP induced the development of LF as a result of promoting hepatic stellate cell (HSC) activation. Therefore, this study revealed unique differential m6A methylation patterns in LF mice and suggested that m6A methylation was associated with the occurrence and course of LF to some extent.

Prevalence and risk factor analysis for the nonalcoholic fatty liver disease in patients with type 2 diabetes mellitus.

ABSTRACT: Although non-alcoholic fatty liver disease (NAFLD) is strongly associated with type 2 diabetes mellitus (T2DM), the diagnosis of NAFLD for T2DM patients remains a challenge.This study aimed to investigate the prevalence and risk factors for the NAFLD in T2DM outpatients.This is a retrospective, cross-sectional study that included 2405 T2DM patients treated and admitted for glucose control into the Endocrinology Department of our hospital from April 2017 to March 2019. Using strict exclusion criteria, the target patients were screened and divided into two groups: NAFLD patients (study group) and non-NAFLD patients (control group). Subsequently, 34 factors were compared between the two groups. Furthermore, multivariate analysis of the NAFLD risk factors was performed using logistic regression. Finally, the diagnostic significance of individual biochemical predictors, as well as the combined predictive indicator (CPI), for NAFLD was estimated using receiver operating characteristic (ROC) curve analysis.In this study, the overall prevalence of NAFLD in T2DM patients was 58.67%. Of the target patients, 17 factors were identified by univariate analysis to be associated with NAFLD, and 8 factors were found to be significant predictors for NAFLD using binary logistic regression modeling. Furthermore, the CPI and C-Peptide represent high diagnostic value for NAFLD in T2DM patients.This study provides a more comprehensive risk factor analysis for NAFLD in T2DM patients. These data can be used to provide timely diagnosis and effective management of NAFLD.

Real-time and rapid quantification of microRNAs in cells and tissues using target-recycled enzyme-free amplification strategy.

In this study, a real-time target-recycled enzyme-free amplification strategy-based test (Trefas test) was developed for rapid, simple, isothermal, and highly sensitive microRNA (miRNA) detection. The Trefas relies on rationally designed sequence-specific hairpins (HPs, HP1 and HP2) and the strand displacement process completely free of environment-susceptible enzymes, enhancing the stability and reproducibility of the test. In the absence of target miRNA, the HP2, modified with a fluorophore and a quencher, maintains stem-loop structure so that the fluorescent signal is quenched. However, in the presence of target miRNA, the target miRNA is repeatedly used to trigger continuous HP1-HP2 hybridizations, restoring fluorescence due to the opening of HP2. The developed miR-21 real-time Trefas test exhibited a broad linear dynamic range of 1 pM to 1 µM and a detection limit of 0.58 pM for miR-21 detection in vitro. In particular, the high specificity of the developed miR-21 real-time Trefas test was prominently exhibited by discriminating single base differences in miRNA sequences. Finally, the expression level of miR-21 in the cell lines and clinical tissues was evaluated by the developed miR-21 real-time Trefas test, and the detection results were highly consistent with the results obtained by stem-loop RT-PCR. In summary, our developed test exhibited great potential for further application in biomedical research and early clinical diagnosis.

A CD44-Targeting Programmable Drug Delivery System for Enhancing and Sensitizing Chemotherapy to Drug-Resistant Cancer.

Programmable drug delivery systems hold great promise to enhance cancer treatment. Herein, a programmable drug delivery system using a chondroitin sulfate (CS)-based composite nanoparticle was developed for enhancing and sensitizing chemotherapy to drug-resistant cancer. The nanoparticle was composed of a cross-linked CS hydrogel shell and hydrophobic cores containing both free drugs and CS-linked prodrugs. Interestingly, the nanoparticle could mediate tumor-specific CD44 targeting. After specific cellular uptake, the payloads were suddenly released because of the decomposition of the CS shell, and the free drug molecules with synergistic effects induced tumor-specific cytotoxicity rapidly. Subsequently, the inner cores of the nanoparticles sustainedly release their cargos in drug-resistant tumor cells to keep the effective drug concentration against the drug efflux mediated by P-glycoprotein. CS dissociated from the outer shell and sensitized cancer cells to the antitumor drugs through downregulation of Bcl-XL, an antiapoptosis protein. Such a programmable drug delivery system with specific tumor-targeting and sensitized therapy is promising for rational drug delivery and provides more versatility for controlled release in biomedical applications.

In vivo transcriptomic analysis of Beauveria bassiana reveals differences in infection strategies in Galleria mellonella and Plutella xylostella.

BACKGROUND: Insect pests have evolved various defense mechanisms to combat fungal infection, and fungi have developed multiple strategies to overcome the immune defense responses of insects. However, transcriptomic analysis of fungal strategies for infecting different pests has not been reported. RESULTS: Transcriptomic profiling of Beauveria bassiana was performed at 12, 24 and 48 h after infecting Galleria mellonella and Plutella xylostella, and 540, 847 and 932 differentially expressed genes were detected, respectively. Functional categorization showed that most of these genes are involved in the ribosome, nitrogen metabolism and oxidative phosphorylation pathways. Thirty-one differentially expressed virulence genes (including genes involved in adhesion, degradation, host colonization and killing, and secondary metabolism) were found, suggesting that different molecular mechanisms were used by the fungus during the infection of different pests, which was further confirmed by disrupting creA and fkh2. Virulence assay results showed that ΔcreA and Δfkh2 strains of B. bassiana had distinct fold changes in their 50% lethal time (LT50 ) values (compared with the control stains) during infection of G. mellonella (ΔcreA: 1.38-fold > Δfkh2: 1.18-fold) and P. xylostella (ΔcreA: 1.44-fold < Δfkh2: 2.25-fold). creA was expressed at higher levels during the infection of G. mellonella compared with P. xylostella, whereas fkh2 showed the opposite expression pattern, demonstrating that creA and Fkh2 have different roles in B. bassiana during the infection of G. mellonella and P. xylostella. CONCLUSION: These findings demonstrate that B. bassiana regulates different genes to infect different insects, advancing knowledge of the molecular mechanisms of Beauveria-pest interactions. © 2018 Society of Chemical Industry.

milR4 and milR16 Mediated Fruiting Body Development in the Medicinal Fungus Cordyceps militaris.

Cordyceps militaris readily performs sexual reproduction, thus providing a remarkably rich model for understanding the processes involved in sexual development. It could regulate expression of human genes by diet-derived miRNA-like RNAs (milRNAs). However, the study of miRNAs in C. militaris has been limited. In the present study, genes encoding Dicers, Argonautes, and RNA-dependent RNA polymerases were identified. Illumina deep sequencing was performed to characterize the milRNAs in C. militaris at asexual and sexual development stages. Total 38 milRNAs were identified and five milRNAs were validated by northern blot and qRT-PCR, out of which, 19 were specific for sexual development. Importantly, the fungi could not form fruiting bodies after disruption of milR4, while the perithecium was formed in advance after over-expression of milR4. Abnormal pale yellow fruiting body primordium, covered with abnormal primordium, was formed in the strain with miR16 disruption. Although no milR4 or milR16 target genes were identified, differential expression of many different genes involved in mycelium growth and sexual development (mating process, mating signaling, and fruiting body development) among these mutants were found. Overall, milRNAs play vital roles in sexual development in C. militaris.

XRN1-associated long non-coding RNAs may contribute to fungal virulence and sexual development in entomopathogenic fungus Cordyceps militaris.

BACKGROUND: Numerous long non-coding RNAs (lncRNAs) identified and characterized in mammals, plants, and fungi have been found to play critical regulatory roles in biological processes. However, little is known about the role of lncRNAs in insect pathogenic fungi. RESULTS: By profiling the transcriptomes of sexual and asexual development in the insect-pathogenic fungus Cordyceps militaris, 4140 lncRNAs were identified and found to be dynamically expressed during fungal development. The lncRNAs had shorter transcript lengths and lower numbers of exons compared to protein-coding genes. The expressed target genes (neighboring and cis-regulated) of various expressed lncRNAs were predicted, and these genes showed significant enrichment in energy metabolism and signaling pathways, such as 'Glycolysis/Gluconeogenesis' and "MAPK signaling pathway". To better understand how lncRNAs function in the fungus, xrn1, the final gene of the NMD pathway, which determines the fate of lncRNAs, was disrupted. The Δxrn1 deletion mutant displayed significant (P < 0.05) attenuation of virulence and a lower growth rate in C. militaris. Quantitative RT-PCR results revealed 10 lncRNAs with significantly higher expression, while 8 of these 10 lncRNA target genes (virulence- and sexual development-related) showed significantly lower expression in Δxrn1 compared to in the wild-type, suggesting that lncRNA expression regulates fungal virulence and sexual development by affecting gene expression. CONCLUSION: These findings suggest that lncRNAs in C. militaris play important roles in the fungal infection progress and fruiting body production, providing a broad repertoire and resource for further studies of lncRNAs. © 2019 Society of Chemical Industry.

The Genome of the Medicinal Macrofungus Sanghuang Provides Insights Into the Synthesis of Diverse Secondary Metabolites.

The mushroom, Sanghuang is widely used in Asian countries. This medicinal fungus produces diverse bioactive compounds and possesses a potent ability to degrade the wood of the mulberry tree. However, the genes, pathways, and mechanisms that are involved in the biosynthesis of the active compounds and wood degradation by Sanghuang mushroom are still unknown. Here, we report a 34.5 Mb genome-encoding 11,310 predicted genes-of this mushroom. About 16.88% (1909) of the predicted genes have been successfully classified as EuKaryotic Orthologous Groups, and approximately 27.23% (665) of these genes are involved in metabolism. Additionally, a total of 334 genes encoding CAZymes-and their characteristics-were compared with those of the other fungi. Homologous genes involved in triterpenoid, polysaccharide, and flavonoid biosynthesis were identified, and their expression was examined during four developmental stages, 10 and 20 days old mycelia, 1 year old and 3 years old fruiting bodies. Importantly, the lack of chalcone isomerase 1 in the flavonoid biosynthesis pathway suggested that different mechanisms were used in this mushroom to synthesize flavonoids than those used in plants. In addition, 343 transporters and 4 velvet family proteins, involved in regulation, uptake, and redistribution of secondary metabolites, were identified. Genomic analysis of this fungus provides insights into its diverse secondary metabolites, which would be beneficial for the investigation of the medical applications of these pharmacological compounds in the future.

MnO2 Nano-Urchin/Graphene Hybrid Electrodes: Facile Synthesis and Enhanced Supercapacitance Performance.

MnO2 with urchin-like nano/micro hierarchical architecture was synthesized through an easy hydrothermal method at low temperature and used to prepare MnO2/graphene hybrid composite as electrode materials through an easy and efficient solution-based method. The MnO2 particles in the composite with 66.7% mass ratio could achieve specific capacitance as high as 451.5 F g(-1) at a scan rate of 10 mV s(-1) and exhibit good cycle stability with 93.8% capacitance retention over 2000 cycles. These properties result from the unique urchin-like nano/micro hierarchical structure of MnO2 as well as the function of graphene in enhancing the conductivity and utilization of MnO2.

A novel peptide inhibitor of platelet aggregation from stiff silkworm, Bombyx batryticatus.

A novel platelet aggregation inhibitory peptide, named BB octapeptide, was isolated from stiff silkworm (Bombyx batryticatus) by gel filtration, anion-exchange, and reverse-phase high performance liquid chromatography. The molecular mass of the peptide was determined to be 885 Da using electrospray ionization mass spectrometry, and the sequence was identified as Asp-Pro-Asp-Ala-Asp-IIe-Leu-Gln using the Edman degradation method. To test its biological activity, the peptide was chemically synthesized using Fmoc solid-phase synthesis method. BB octapeptide inhibited rabbit platelet aggregation that was induced by collagen and epinephrine, with the IC50 values of 91.14 µM and 104.50 µM, respectively. After intravenous administrated in mice (30 mg/kg, 4 days), BB octapeptide showed similar ex vivo efficacy of inhibiting platelet aggregation as aspirin (10 mg/kg). In addition, this peptide prevented paralysis and death in pulmonary thromboembolism model and significantly reduced ferric chloride-induced thrombus formation in rats. Moreover, it exhibited low cytotoxicity in a cellular model. In conclusion, this is the first report that a novel platelet aggregation inhibitory peptide was isolated from stiff silkworm (B. batryticatus). Due to the excellent efficacy in reducing platelet aggregation and low toxicity, it can be a valuable lead compound for new drug design and development.