Optimization of Hydrolysis Conditions for the Isolation of Angiotensin-I Converting Enzyme(ACE) Inhibitory Peptides from Rhopilema hispidum

To obtain the maximum angiotensin-I converting enzyme(ACE) inhibitory activity, the protein hydrolysis conditions of the jellyfish Rhopilema hispidum were optimized using response surface methodology(RSM). Trypsin was selected to produce R. hispidum protein hydrolysates(RPH) with ACE inhibitory activity. The optimal parameters for producing protein hydrolysates with the highest ACE inhibitory activity were as follows: hydrolysis time 5 h, hydrolysis temperature 50℃, and the enzyme-to-substrate ratio 6%. Under these conditions, the ACE inhibitory rate of RPH could reach 64.28% ± 5.72%. In addition, RPH contained high levels of Gly, Glu, Pro, Ala, Asp and Arg, with a molecular weight distribution range of 0.32–6.84 kDa. The following three novel ACE inhibitory peptides were isolated and identified: Ile-Gly-Glu-Thr-Gly-Pro, Gly-Ala-Thr-Gly-Pro-Ala-Gly-Tyr-Val and Gly-AlaPhe-Gly-Pro-Gly-Gly-Leu-Val-Gly-Arg-Pro. The IC_(50) values of the ACE inhibitory activity of these three purified peptides were 19.07, 27.42 and 31.26 μmol L^(-1), respectively. These results suggested that proteins and peptides isolated from R. hispidum could be utilized as antihypertensive functional food sources.

Identifi cation and characterization of the bZIP transcription factor family in yellowhorn

The basic leucine zipper(bZIP)transcription factor family is one of the largest and most diverse families in plants,regulating plant growth and development and playing an essential role in response to abiotic and biotic stresses.However,little is known about the biological functions of bZIP proteins in yellowhorn(Xanthoceras sorbifolium).Recently,64 XsbZIP genes were identifi ed in the yellowhorn genome and found to be disproportionately distributed in linkage groups.The XsbZIP proteins clustered into 11 groups based on their phylogenetic relationships with AtbZIP,ZmbZIP and GmbZIP proteins.Five intron patterns in the basic and hinge regions and additional conserved motifs were defi ned,both supporting the group classifi cation and possibly contributing to their functional diversity.Compared to tandem duplication,the segment duplication greatly contributed to the expansion of yellowhorn bZIP genes.In addition,most XsbZIP genes harbor several stress responsive cis-elements in their promoter regions.Moreover,the RNA-seq and qRT-PCR data indicated XsbZIP genes were extensively involved in response to various stresses,including salt(NaCl),cold and abscisic acid,with possibly diff erent molecular mechanisms.These results provide a new understanding of the biological functions of bZIP transcription factors in yellowhorn.

A culture-free method for detection of Vibrio vulnif icus from coastal seawater based on loop-mediated isothermal amplification targeting vcgC gene

Vibrio vulnificus is an opportunistic pathogen widely distributed in estuarine and coastal seawaters.In this study,a culture-free method was developed to rapid detection of V.vulnificus in all seasons,based on loop-mediated isothermal amplification(LAMP) targeting virulent-correlated gene(vcg).The new assay method allows differentiation between the virulent and non-virulent strain of V.vulnificus accurately.This method also allows effective detection of the pathogeny in winter when the bacterium lives in the viable but non-culturable(VBNC) state.A total of 30 costal seawater samples collected in all seasons were used for the evaluation of this method.The results show that the method is sensitive,accurate and convenient.

The structure of integument and wax glands of Phenacoccus fraxinus(Hemiptera:Coccoidea:Pseudococcidae)

Using scanning electron microscopy and optical microscopy,we studied the structure of the integument and wax glands of the mealybug,Phenacoccus fraxinus Tang(Hemiptera:Coccoidea:Pseudococcidae).We observed the ultrastructure of four wax pores including trilocular,quinquelocular,and multilocular pores as well as tubular ducts,recording characteristics of their structure,size and distribution.We found that that the integument of the mealybug consists of three main layers-the procuticle,epidermis and basement membrane-and four sub-layers of the procuticle-the epicuticle,exocuticle,endocuticle and formation zone.The waxsecreting gland cells were closely arranged in epidermis.All of them were complex and composed of one central cell and two or more lateral cells.These complex cells possess a large common reservoir for collection and storage.Synthesized by the glandular cells,the wax is excreted outside integument through canals.

Effects of Selenium on Fusarium Growth and Associated Fermentation Products and the Relationship with Chondrocyte Viability

这研究在镰刀霉紧张的生长和在 chondrocytes 损害的相关指示物上从真菌的文化提取的产品的效果上决定了硒的效果。结果证明硒补充在紧张的菌丝生长上导致了微分效果。chondrocyte 损害指示物铺平，包括房间生存能力， proteoglycan 和类型 II 骨胶原内容和他们的 mRNA 表情，当 chondrocytes 与发酵摘录被孵化时，都被归结为改变度，禁止的效果变化了取决于补充到真菌的文化媒介的硒内容。结果显示某些链关系在环境，由真菌的一些代谢物的生产，和 chondrocyte 损坏的出现在硒的内容之间存在。它进一步在 Kaschin 山涧疾病致病优点玩的这种关系和角色的程度学习。

The structure of integument and wax glands of Phenacoccus fraxinus(Hemiptera:Coccoidea:Pseudococcidae)

Using scanning electron microscopy and optical microscopy,we studied the structure of the integument and wax glands of the mealybug,Phenacoccus fraxinus Tang(Hemiptera:Coccoidea:Pseudococcidae).We observed the ultrastructure of four wax pores including trilocular,quinquelocular,and multilocular pores as well as tubular ducts,recording characteristics of their structure,size and distribution.We found that that the integument of the mealybug consists of three main layers-the procuticle,epidermis and basement membrane-and four sub-layers of the procuticle-the epicuticle,exocuticle,endocuticle and formation zone.The wax-secreting gland cells were closely arranged in epidermis.All of them were complex and composed of one central cell and two or more lateral cells.These complex cells possess a large common reservoir for collection and storage.Synthesized by the glandular cells,the wax is excreted outside integument through canals.

Annotation of miRNAs in the COVID-19 Novel Coronavirus

The coronavirus disease 2019(COVID-19)coronavirus is a new strain of coronavirus that had not been previously detected in humans.As its severe pathogenicity is concerned,it is important to study it thoroughly to aid in the discovery of a cure.In this study,the microRNAs(miRNAs)of COVID-19 were annotated to provide a powerful tool for the study of this novel coronavirus.We obtained 16 novel coronavirus genome sequences and the mature sequences of all viruses in the microRNA database(miRbase),and then used the miRNA matures sequences of the virus to perform the Basic Local Alignment Search Tool(BLAST)analysis in the coronavirus genome,extending the matched regions of approximately 20 bp to two segments by 200 bp.Six sequences were obtained after deleting redundant sequences.Then,the hairpin structures of the mature miRNAs were determined using RNAfold.The mature sequence on one hairpin arm was selected into a total of 4 sequences,and finally the relevant miRNA precursor prediction tools were used to verify whether the selected sequences are miRNA precursor sequences of the novel coronavirus.The miRNAs of the novel coronavirus were annotated by our newly developed method,which will lay the foundation for further study of this virus.

The Pathogen and Wound Induces Expression of Genes Related to Proanthocyanidins (PAs) Synthesis in Cotton Leaves

Responses to biotic and abiotic stress have been extensively studied in plants. In the current proteomic study, the cotton (Gossypium hirsutum L.) seedlings were infected with Verticillium dahliae by root-dip inoculation using suspension of fungal conidia. The different proteins were analyzed by two-dimensional gel elactrophoresis (2-DE), and flavanone 3-hydroxylase (F3H) showed a significantly up-regulation in cotton leaf after V. dahliae infection. Further research revealed F3H and the downstream genes of F3H in proanthocyanidins (PAs) biosynthesis were also significantly induced and showed coordinate expression patterns during wounding. The results indicate that PAs in cotton act an important role in response to infection V. dahliae and wounding.

Chemical constituents from Schisandra sphenanthera

五味子 sphenanthera 的茎的化学成分第一次被描述。这调查导致了新酉分的 glycoside (1 ) 的隔离，与七已知的混合物一起。1 的结构被使用分光镜的技术分配，包括 2D NMR 系列。

Effect of oridonin-mediated hallmark changes on inflammatory pathways in human pancreatic cancer(BxPC-3) cells

AIM:To investigate the effect of oridonin on nuclear transcription factors and to study the relationship between biological behavior and inflammatory factors in human pancreatic cancer(BxPC-3)cells.METHODS:BxPC-3 cells were treated with various concentrations of oridonin,and viability curves were generated to test for inhibitory effects of the drug on cells.The expression of cytokines such as interleukin-1β(IL-1β),IL-6,or IL-33 was detected in BxPC-3 cell supernatants using an enzyme-linked immunosorbent assay(ELISA),and the protein expression of nuclear transcription factors including nuclear factorκB,activating protein-1,signal transducer and activator of transcription 3,bone morphogenetic protein 2,trans-forming growth factorβ1 and sma and mad homologues in BxPC-3 cells was detected using Western blot.Carcinoma hallmark-related proteins such as survivin,vascular endothelial growth factor,and matrix metallopeptidase 2 were also detected using immunoblotting,and intra-nuclear IL-33 expression was detected using immunofluorescent staining.RESULTS:Treatment with oridonin reduced the viability of BxPC-3 cells in a dose dependent manner.The cells exhibited reduced growth following treatment with 8μg/mL oridonin(13.05%±3.21%,P<0.01),and the highest inhibitory ratio was 90.64%±0.70%,which was achieved with oridonin at a dose of32μg/mL.The IC50 value of oridonin in BxPC-3 cells was 19.32μg/mL.ELISA analysis revealed that oridonin down-regulated the inflammatory factors IL-1β,IL-6,and IL-33 in a dose-dependent manner.IL-1βexpression was significantly reduced in the 16 and 32μg/mL treatment groups compared to the control group(12.97±0.45 pg/mL,11.17±0.63 pg/mL vs 14.40±0.38pg/mL,P<0.01).Similar trends were observed for IL-6expression,which was significantly reduced in the 16and 32μg/mL treatment groups compared to the control group(4.05±0.14 pg/mL vs 4.45±0.43 pg/mL,P<0.05;3.95±0.13 pg/mL vs 4.45±0.43 pg/mL,P<0.01).IL-33 expression was significantly reduced in the8,16,and 32μg/mL treatment groups compared to the control group(911.05±14.18 pg/mL vs 945.25±12.09 pg/mL,P<0.05;802.70±11.88 pg/mL,768.54±10.98 pg/mL vs 945.25±12.09 pg/mL,P<0.01).Western blot and immunofluorescent staining analyses suggested that oridonin changed the hallmarks and regulated the expression of various nuclear transcription factors.CONCLUSION:The results obtained suggest that oridonin alters the hallmarks of pancreatic cancer cells through the regulation of nuclear transcription factors.

Cloning and molecular characterization of△^(12)-fatty acid desaturase gene from Mortierella isabellina

瞄准：克隆三角洲(12 ) Mortierella isabellina 并且到的丰满的酸 desaturase 基因机能上地描绘这基因在试管内和在活体内。方法：反向的 transcriptional 聚合酶链反应(RT-PCR ) 被用来克隆三角洲(12 ) 的开的读物框架 Mortierella isabellina 的丰满的酸 desaturase 基因(D12D ) 。Plasmids pEMICL12 和 pYMICL12 与它被构造。pEMICL12 被转变成埃希氏杆菌属关口 i (E.coli ) 为在有 IPTG 的正式就职以后的表示的紧张 BL21 使用 CaCl (2 ) 方法。pTMICL12 在半乳糖的正式就职下面为表示用锂醋酸盐方法被转变成酿酒酵母紧张 INVSc1。北弄污方法被用来在 S.cerevisiae 紧张 INVSc1 在这基因的 transcriptional 水平上调查温度的效果。结果：Recombinant 原生质标志 pEMICL12 和 pTMICL12 成功地被构造并且与适当方法独立转变了成 E.coli 和 S.cerevisiae。在有 IPTG 和半乳糖的正式就职以后，它被发现三角洲(12 ) 的那表情在 E.coli 和 S 的丰满的酸 desaturase 基因。在导致的适当条件下面的啤酒活跃三角洲(12 ) 的生产丰满的酸 desaturase，它能由 GCMS 察觉在试管内和在活体内分别地把 17.876% 油酸和 17.604% 变换成亚油酸。结论：克隆并且在 E.coli 和 S.cerevisiae 的 M.isabellina D12D 基因的表示成功地被完成。

Schisanlactone G,a new 3,4-seco-lanostane triterpenoid from Schisandra sphenanthera

新 3,4-seco-lanostane triterpenoid， schisanlactone G (1 ) ，从五味子 sphenanthera 的水果被孤立。它的结构根据广泛的分光镜的分析被建立。

Identification of yellowhorn(Xanthoceras sorbifolium)WRKY transcription factor family and analysis of abiotic stress response model

WRKY transcription factors are widely distributed in higher plants and play important roles in many biological processes,including stress resistance.The recently published genome sequence of yellowhorn,an oil tree with robust resistance to cold,drought,heat,salt and alkali,provides an excellent opportunity to identify and characterize the entire yellowhorn WRKY protein family and a basis for the study of abiotic stress resistance of WRKY gene family in forest species.In the present comprehensive analysis of WRKY transcription factors in yellowhorn,65 WRKY genes were identified and defined based on their location on the chromosome.According to their structure and phylogenetic relationships,XsWRKY genes clustered into WRKY groupsⅠ-Ⅲ.Segmental duplication events played a significant role in the expansion of WRKY gene family.Furthermore,transcriptomic data and real-time quantitative PCR analysis showed that expression of XsWRKY genes responding to salt and drought stresses and a hormone treatment.We also determined structures of the encoded proteins,c is-elements of the promoter region,and expression patterns.These results provide a foundation for the study of the biological function of WRKY transcription factors in yellowhorn.

The chemical constituents from Schisandra propinqua var. intermedia

新苯乙烯的酸 monoterpenoid 酉旨， intermedin C (1 ) ，从五味子 propinqua var 的天线部分被孤立。intermedia。1 的结构被包括广泛的 1D 和 2D NMR 技术的分光镜的方法阐明。

Study on the mechanism of SND in the treatment of anxiety disorders based on network pharmacology, molecular docking and experiment validation

Background:Sini decoction(SND)is a classic traditional Chinese medicine(TCM)formulation that can be used to treat anxiety-related disorders,but the active substance and underlying molecular mechanism of its anxiolytic effects are unknown.In this study,network pharmacology,molecular docking research and experimental verification methods were used to preliminarily explore the bioactive compounds and potential target mechanisms of SND anxiolytic.Methods:The active components and corresponding targets of SND were collected by TCMSP.GeneCards,OMIM,PharmGkb,TTD and Drugbank were used to search for the targets of anxiety disorders.The core target of SND in the treatment of anxiety was screened by PPI.R language was used to analyze the intersection targets of SND in the treatment of anxiety disorders by GO and KEGG enrichment analysis.AutoDock Vina was used for molecular docking,and Discovery Studio was used for visual conformation analysis after docking.The anti-anxiety effect and molecular mechanism of SND were studied by in vivo experiment.Results:Based on network pharmacological analysis,we obtained 112 active ingredients and 350 effective targets related to anxiety from SND.In PPI analysis,26 targets such as STAT3,MAPK3,MAPK1,MAPK14,SRC,HSP90AA1,TP53 and PIK3CA were identified as core targets.GO and KEGG analysis showed that the anxiolytic mechanism of SND may be related to the neuroactive ligand-receptor interaction pathway and inflammatory pathway.Molecular docking showed that quercetin,naringenin,licochalcone A had high affinity with JAK2,MAPK14 and MAPK3.Animal experiments have shown that SND reverses the upregulation of GluN2B(NMDAR)and GluA1(AMPAR)proteins,and SND improves anxiety disorders by regulating glutamate transmitter levels,which may be related to neuroactive ligand-receptor interaction pathways,particularly glutamate receptors.Conclusion:This study shows that SND can improve FS-induced behavioral changes in mice and can modulate hippocampal synapse-associated protein defects,partially reversing glutamate receptor expression through the neuroactive ligand-receptor interaction pathway,and further improved anxiety disorders.At the same time,combined with network pharmacology and molecular docking,the key components,core targets and related pathways of SND are discussed,which shows that the active components of SND play an effective role in anxiety through multi-targets and multi-pathways,which provides a reference for the material basis and mechanism of SND.

Super-assembly of integrated gold magnetic assay with loopmediated isothermal amplification for point-of-care testing

With the increasing global threat of various diseases and infections,it is essential to develop a fast,low-cost,and easy-to-use point-of-care testing(POCT)system for inspections at all levels of medical institutions and self-examination at home.In this work,gold magnetic nanoparticles(GMNPs)are used as the key material,and a rapid visual detection method is designed through integrating loop-mediated isothermal amplification(LAMP)and lateral flow assay(LFA)biosensor for detecting a variety of analytes which includes whole blood,buccal swabs,and DNA.It is worth to note that the proposed method does not need DNA extraction.Furthermore,uracil DNA glycosylase(UDG)is employed to eliminate carrier contamination for preventing false positive results.The whole detection process can be finished within 25 min.The accuracy of detection is measured by assessing the polymorphisms of the methylenetetrahydrofolate reductase(MTHFR)C677T.The detection limit of the newly developed extraction-free detection system for MTHFR C677T is 0.16 ng/μL.A preliminary clinical study of the proposed method is carried out by analyzing 600 clinical samples(including 200 whole blood samples,100 buccal swabs,and 300 genomic DNA samples).The results indicate that the proposed method is 100%consistent with the sequencing results which provides a new choice for POCT and shows a broad application prospect in all levels of medical clinics and at home.

Synthesis and anti-HIV-1 activity of the conjugates of gossypol with oligopeptides and D-glucosamine

A series of novel gossypol derivatives were synthesized and screened for their in vitro anti-HIV-1 activity.The results showed that replacing the aldehyde groups of gossypol with certain oligopeptides and Dglucosamine not only reduced the cytotoxicity of gossypol derivatives but also enhanced their antiviral activity against HIV-1. Interestingly, D-glucosamine derivative of gossypol that lacked the COONa group also exhibited the same potent anti-HIV-1 activity as oligopeptide derivatives with the COONa group.These compounds blocked the entry of HIV-1IIIBinto target cell, which was similar to T20. Furthermore,the molecular docking analysis rationalized their anti-HIV-1 activity. The results also implied that certain oligopeptides and D-glucosamine were important moities to prepare gossypol derivatives as HIV-1 entry inhibitors besides certain amino acids.