New Cytotoxic Salinosporamides from the Marine Actinomycete Salinisporatropica

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北玄参根中的一个新环烯醚萜衍生物_英文_

A new iridoid derivative from the roots ofScrophularia buergerianaWU Xi-min 1†, ZHANG Liu-qiang 1†, CHEN Xiao-chong 1, FENG Li 1,XING Wang-xing 2*, LI Yi-ming 1*(1. School of Pharmacy, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China;2. School of Medicine, Hangzhou Normal University, Hangzhou 310036, China )Abstract : Phytochemical investigation of the roots of Scrophularia buergeriana Miq. (Scrophulariaceae), resulted in the isolation of a new iridoid derivative named as buergerinin (1). Its structure was elucidated as rel-(1R , 5R , 6R )-(2-oxa-bicyclo[3.3.0]oct-7-en-6, 7-diyl)dimethoxypropane based mainly on MS and 1D and 2D NMR spectroscopic analyses.Key words : Scrophularia buergeriana ; iridoid derivative; buergerinin CLC number : R284 Document code : A Article ID : 0513-4870 (2014) 07-1019-03北玄参根中的一个新环烯醚萜衍生物吴喜民1†, 张刘强1†, 陈小冲1, 冯丽1, 邢旺兴2*, 李医明1*(1. 上海中医药大学中药学院, 上海 201203; 2. 杭州师范大学医学院, 浙江杭州 310036)摘要: 从北玄参块根中发现了一个新的环烯醚萜衍生物, 命名为buergerinin (1)。

Anthocyanidin reductases from Medicago truncatula

Anthocyanidin reductases from Medicago truncatulaand Arabidopsis thalianaDe-Yu Xie,Shashi B.Sharma,and Richard A.Dixon *Plant Biology Division,Samuel Roberts Noble Foundation,2510Sam Noble Parkway,Ardmore,OK 73401,USAReceived 19November 2003,and in revised form 5December 2003AbstractAnthocyanidin reductase (ANR),encoded by the BANYULS gene,is a newly discovered enzyme of the ﬂavonoid pathway involved in the biosynthesis of condensed tannins.ANR functions immediately downstream of anthocyanidin synthase to convert anthocyanidins into the corresponding 2,3-cis -ﬂavan-3-ols.We report the biochemical properties of ANRs from the model legume Medicago truncatula (MtANR)and the model crucifer Arabidopsis thaliana (AtANR).Both enzymes have high temperature optima.MtANR uses both NADPH and NADH as reductant with slight preference for NADPH over NADH.In contrast,AtANR only uses NADPH and exhibits positive cooperativity for the co-substrate.MtANR shows preference for potential anthocyanidin substrates in the order cyanidin >pelargonidin >delphinidin,with typical Michaelis–Menten kinetics for each substrate.In contrast,AtANR exhibits the reverse preference,with substrate inhibition at high concentrations of cyanidin and pelargonidin.(+)-Catechin and (Æ)-dihydroquercetin inhibit AtANR but not MtANR,whereas quercetin inhibits both enzymes.Possible catalytic reaction sequences for ANRs are discussed.Ó2003Elsevier Inc.All rights reserved.Keywords:Condensed tannins;Flavonoids;Anthocyanidin;Kinetics;Model legumeCondensed tannins (CTs)1are oligomers or poly-mers of ﬂavonoid units (ﬂavan-3-ols,also known as catechins)linked by single C 4–C 8or C 4–C 6carbon–carbon bonds (Fig.1A).CTs occur widely throughout the plant kingdom,often in seed coats but also in other tissues such as leaves,ﬂowers,and stems.They play protective functions in the plant,particularly against herbivores [1,2],and have recently attracted consider-able attention in view of their potentially beneﬁcial eﬀects on human and animal health [1–3].CTs bind reversibly to proteins,and the tannins present in the leaves of some forage legumes such as Desmodium uncinatum and Lotus corniculatus can thereby slow the rate of protein degradation in the rumen and thus protect cattle and sheep from pasture bloat [1,3].CTsare also powerful antioxidants,and this property may explain their beneﬁcial eﬀects on cardiac health and immunity [4,5].The ﬂavan-3-ol building blocks of CTs are also antioxidants [6,7],and clinical studies have shown protective eﬀects of catechin,epicatechin,gal-locatechin,and epigallocatechin against stomach can-cer [8].It has recently been shown that epicatechin present in dark chocolate has potential to promote cardiovascular health in humans [9].CTs share the same upstream biosynthetic pathway as the anthocyanin ﬂower pigments (Fig.1A),and this pathway has been well deﬁned at both the biochemical and molecular genetic levels [10,11].It was previously believed that the branch point between the CT and an-thocyanin pathways occurred at the level of leucoanth-ocyanidin,which is converted to anthocyanidin by the action of leucoanthocyanidin dioxygenase (also known as anthocyanidin synthase,ANS)and,hypothetically,to ﬂavan-3-ol by a leucoanthocyanidin reductase (LAR).However,this model,which did not explain the origin of the 2,3-cis stereochemistry predominant in the building blocks of many CTs,has recently undergone revision as*Corresponding author.Fax:1-580-224-6692.E-mail address:radixon@ (R.A.Dixon).1Abbreviations used:CT,condensed tannin;Mt,Medicago trun-catula ;At,Arabidopsis thaliana ;ANR,anthocyanidin reductase;ANS,anthocyanidin synthase;LAR,leucoanthocyanidin reductase;DHQ,dihydroquercetin;MBP,maltose-binding protein;MDAR,monode-hydroascorbate reductase;DFR,dihydroﬂavonol reductase.0003-9861/$-see front matter Ó2003Elsevier Inc.All rights reserved.doi:10.1016/j.abb.2003.12.011Archives of Biochemistry and Biophysics 422(2004)91–102ABB/locate/yabbia result of the biochemical characterization of the BANYULS gene[12].The BANYULS gene was named after the color of a French red wine for a mutant of Arabidopsis thaliana that lacked CTs in the seed coat endothelium and pre-cociously accumulated anthocyanins[13,14].It was originally proposed that BANYULS might encode LAR [14].However,when expressed in Escherichia coli,the product of a BANYULS ortholog from the model le-gume Medicago truncatula eﬃciently convertedantho-cyanidins into their correspondingﬂavan-3-ols in the presence of NADPH[12].This resulted in the proposal of a new pathway to CTs in which anthocyanidins were key intermediates,as substrates for the BANYULS gene product anthocyanidin reductase(ANR).Chemical characterization of the reaction products of ANR identiﬁed two isomers ofﬂavan-3-ol,namely2R,3R-2, 3-cis-and2S,3R-2,3-trans[12].With cyanidin as sub-strate,these products are())-epicatechin and())-cate-chin,respectively.The2R,3R-2,3-cis-ﬂavan-3-ol,which was the major reaction product,is the common building block of CTs in many plants including Arabidopsis[15] and alfalfa(Medicago sativa)[16].Recently,a LAR gene has been cloned following puriﬁcation of the corresponding enzyme from D.un-cinatum leaf tissue and shown to be a member of the isoﬂavone reductase(IFR)-like gene family.The enzyme converted radiolabeled leucocyanidin to(+)-catechin (2R,3S-2,3-trans-ﬂavan-3-ol)[17].(+)-Catechin is often the starter unit for CT polymerization.Together,ANR and LAR would therefore appear to be suﬃcient for formation of most monomeric building blocks of CTs. However,still very little is understood concerning the biological processes resulting in CT polymerization,or the reaction mechanisms whereby the building blocks are produced via ANR and LAR.The basic anthocyanidin structure is generally con-sidered to be the coloredﬂavylium cation or salts thereof (Fig.1B)[18–22].However,anthocyanidins are not sta-ble in aqueous solution and undergo rapid,pH-depen-dent,reversible transformations(Fig.1B)[18,22,23].The colors fade rapidly in weakly acidic solutions(pH3–5), presumably due to formation of an equilibrium mixture of theﬂavylium cation and the colorless chromen-2-ol pseudobase(Fig.1B)[22].At pH5–8,anthocyanidins can also rapidly transform to the anhydro base or quinoidal base(Fig.1B)[22,23].These transformations complicate the assignment of a reaction mechanism to ANR.We here present a comparative study of the bio-chemical properties of ANRs from M.truncatula and A. thaliana.A full understanding of the mode of catalysis by ANR will facilitate future attempts to engineer con-densed tannins or catechins in a variety of plant species for improving nutritional and health-related traits.In-terestingly,the ANR enzymes from these two sources have quite diﬀerent substrate speciﬁcities and kinetic properties.We present hypothetical mechanisms for the ANR reaction.Materials and methodsChemicalsCyanidin chloride,pelargonidin chloride,and delphinidin chloride were purchased from Indoﬁne (Hillsborough,NJ,USA).Ten mM methanol stock so-lutions were stored at)20°C.NADPH and NADH were from Sigma(St.Louis,MO)and solutions were freshly prepared in cold reaction buﬀer when used for enzymatic assay.(+)-Catechin,(Æ)-catechin,())-epi-catechin,())-gallocatechin,())-epigallocatechin,(Æ)-di-hydroquercetin(DHQ),and quercetin were from Sigma, and methanolic stock solutions were stored at)20°C.Expression of MtANR and AtANR in E.coliThe open reading frame of the MtANR gene was ampliﬁed by polymerase chain reaction(PCR)using pSE380-BAN plasmid as template[12].The amino terminal primer,50-GCGGCGGAATTCATGGCTAG TATCAAACAAATAG-30containing the ATG start codon,introduced an Eco RI restriction site(under-lined),and the carboxyl terminal primer50-GCGGCG TCTAGATCACTTGA TCCCCTGAGTCTTC-30,in-cluding the stop codon,introduced an Xba I restriction site.PCR was carried out using Taq DNA polymerase (Promega,Madison,WI)at94°C for2min,35cycles of 94°C for45s,60°C for45s,and72°C for1.5min; followed by a10min extension at72°C.The PCR fragment was gel puriﬁed using a Sephaglas Bandprep kit(New England Biolabs,Beverly,MA)and ligated into pGEM-T-easy vector(Promega).The pGEM-T-MtANR plasmid was transformed into E.coli DH5a competent cells for sequence analysis.The open reading frame was then excised by digestion with Eco RI and Xba I and re-ligated into Eco RI and Xba I digested pMAL-c2X vector as an N-terminal fusion to the maltose-binding protein(MBP)gene to generate the protein expression construct pMMtANR.To clone AtANR for protein expression,total RNA from young siliques of A.thaliana cv Columbia was isolated using the Tri-Reagent kit(Molecular Re-search Center,Cincinnati,OH).First strand cDNA was synthesized from4l g total RNA from siliques by Su-perscriptII reverse transcriptase(Invitrogen,Carlsbad, CA)according to the manufacturerÕs instructions.The ANR gene was ampliﬁed from10l l of theﬁrst strand cDNA by highﬁdelity Pfu DNA polymerase-mediated PCR using forward primer50-GGGCCCATGGAC CAGA CTCTTACACACACCGGA-30and reverse primer50-CCCAGATCTAGAATGAGACCAAAG ACTCATAT ACT-30at94°C for5min,30cycles of 94°C for1min,55°C for1min,and72°C for1.5min, followed by a further10min extension at72°C.The PCR fragment was digested with Nco I and Xba I,and cloned into Nco I–Xba I digested pRTL2to give the construct pSBP77,which was then sequenced.For ex-pression of AtANR in E.coli,pSBP77was digested with Nco I and then blunted with mung bean nuclease(New England BioLabs).The ANR coding region fragment was puriﬁed after digestion with Xba I,and ligated intoD.-Y.Xie et al./Archives of Biochemistry and Biophysics422(2004)91–10293Xmn I and Xba I digested pMAL-c2X as an N-terminal fusion to the MBP gene to generate the protein expression construct pMAtANR.pMMtANR,pMAtANR,and pMAL-c2X(as control vector)were transformed into NovaBlue(DE3) competent cells(Novagen,Madison,WI).A single col-ony harboring either construct or control vector was inoculated into1liter of LB broth containing100mg/ liter ampicillin and cells were grown to an OD600of0.3 at37°C and250rpm.The cells were then transferred to 16°C and grown to an OD600of0.6–0.7,at which point isopropyl-1-thio-b-D-galactopyranoside(IPTG)was added to aﬁnal concentration of1mM.The cells were harvested24h later by centrifugation at3000rpm at 4°C for15min.Pellets were either used directly for enzyme extraction or stored at)80°C.Enzyme extraction and puriﬁcationAll procedures were performed at4°C.Crude en-zyme was extracted as described previously[12]and further puriﬁed on an amylose column.Three to4ml amylose homogenate(New England Biolabs)was loa-ded into a2.5Â16cm column,and then washed and equilibrated with at least10volumes of washing buﬀer (20mM Tris–HCl,pH7.0,and10mM b-mercap-toethanol).The crude extract was dilutedﬁve times with washing buﬀer and then loaded onto the column at aﬂow rate of1ml/min.The column was washed with at least15column volumes of washing buﬀer and the MBP-ANR fusion protein eluted with washing buﬀer supplemented with10mM maltose.The fusion protein was concentrated to 1.5–10l g/l l by passing through a10,000molecular weight cut-oﬀAmicon Ultra(Millipore)membrane and then analyzed by electrophoresis on a12%SDS–polyacrylamide gel stained with0.25%Coomassie brilliant blue(R250). ANR protein was separated from MBP by protease cleavage with Factor Xa(New England Biolabs)fol-lowing the manufacturerÕs protocol.Enzyme assays and determination of kinetic parameters All enzyme assays were performed by measuring ap-pearance ofﬂavan3-ol products by HPLC after stop-ping reactions after the required time periods.For determination of temperature optimum,enzyme assays were carried out for30min in a total volume of200l l containing100mM Tris–HCl,pH7.0,100l M cyanidin chloride,2mM NADPH,and50–100l g MBP-ANR or 30l g puriﬁed cleaved ANR.Determination of pH de-pendence and reaction kinetics was performed with MBP-ANR fusion proteins.pH proﬁles for MtANR and AtANR were determined at their respective temperature optima of45°C(MtANR) and55°C(AtANR)with30min incubations in aﬁnal volume of200l l consisting of50mM citrate/phosphate buﬀer,pH4.0–7.0,50mM Mes buﬀer,pH5.0–7.0or 100mM Tris–HCl buﬀer,pH7.0–8.8,100l M cyanidin, 2mM NADPH,and50–100l g puriﬁed fusion protein.Kinetic constants for NADPH or NADH were de-termined at the respective temperature optima of MtANR and AtANR in a total volume of200l l con-taining50mM Mes buﬀer,pH6.0(for MtANR)or 100mM Tris–HCl,pH8.0(for AtANR),100l M cyanidin,0–12mM NADPH or NADH,and100l g MBP-fusion protein,with30min incubation.Kinetic constants for anthocyanidin substrates were likewise determined in a total volume of200l l containing100l g MBP-fusion protein,0–300l M cyanidin chloride or delphinidin chloride,or0–500l M pelargonidin chlo-ride,and2mM NADPH.To determine the eﬀects ofﬂavonoids and Naþions, enzyme assays were carried out at the respective tem-perature and pH optima in a total volume of200l l containing100l g MBP-fusion protein,100l M cyanidin chloride,2mM NADPH,and either(Æ)-DHQ(0–1mM),(+)-catechin(0–1mM),quercetin(0–0.1mM)or NaCl(0–400mM).To determine whether MtANR or AtANR can epi-merize or racemize theﬂavan-3-ol products,assays were carried out at the respective temperature and pH optima in a total volume of200l l containing100l g fusion protein,2mM NADPH,and35–70l M())-epicatechin or50–100l M(Æ)-catechin in the presence or absence of 100l M cyanidin.Control experiments were performed under the same conditions with buﬀer or boiled enzyme instead of active enzyme.All enzyme reactions were initiated by adding enzyme and stopped by adding1ml ethyl acetate with vigorous vortexing for1min,and then centrifuged for1min at 10,000rpm.The ethyl acetate supernatant phase(0.9ml) was transferred to a new1.5ml microcentrifuge tube and dried under nitrogen gas at room temperature. Residues were dissolved in50l l HPLC grade methanol for HPLC analysis.HPLC analysis and data integrationHPLC analysis used a HP1100machine and C18 reversed phase column with UV detection at280nm for ())-epicatechin,(Æ)-catechin,(+)-catechin,())-epiafz-elechin,and())-afzelechin,and at270nm for())-gal-locatechin and())-epigallocatechin.The solvent system consisted of water(solvent A)and acetonitrile(solvent B).The gradient program consisted of ratios of solvent A to solvent B of95:5(0–5min),95:5to93:7(5–7min), 93:7(7–25min),93:7to60:40(25–40min),60:40to5:95 (40–40.5min),and5:95(40.5–49.5min).Forty l l sam-ples was injected with aﬂow rate of1.5ml per min. Product peaks were identiﬁed by retention time and UV spectrum as previously described[12].Authentic94 D.-Y.Xie et al./Archives of Biochemistry and Biophysics422(2004)91–102standards of(Æ)-catechin,())-epicatechin,())-gallo-catechin,and())-epigallocatechin were injected for reference.Each experiment was performed in triplicate and re-peated at least once.Product formation was estimated by integrating peak area using HP Chemstation Soft-ware.Amounts of reaction products were calculated based on established standard curves using authentic samples.Due to lack of available standards of())-epi-afzelechin and())-afzelechin,the products produced from pelargonidin were simply estimated by integrating peak areas.Values could not be calculated with pe-largonidin as substrate.The enzymatic products con-sisted of two isomers.Initial reaction velocity was calculated as the total sum of the products,i.e., ())-catechin and())-epicatechin,())-afzelechin and ())-epiafzelechin,or())-gallocatechin and())-epigallo-catechin.K m,V max,½S0:5,and n H values were calculatedthrough various plots of initial reaction velocity versus substrate concentration.V max values were not calculated for pelargonidin due to lack of authentic standards of (epi)-afzelechin for absolute quantitation.ResultsSequence comparison of MtANR and AtANR MtANR and AtANR(from A.thaliana ecotype Columbia)share only60%amino acid sequence identity. MtANR consists of338amino acids with a molecular weight(MW)of36,977Da,whereas AtANR consists of 340amino acids,with an MW of37,904Da.Both MtANR and AtANR contain the classical Rossmann dinucleotide-binding domain[24]character-ized by a conserved glycine rich amino acid sequence ﬁngerprint of either GXGXXA(such as found in monodehydroascorbate reductases,MDAR)[25]or GXXGXXG(such as found in plant dihydroﬂavonol reductases,DFR)(Fig.2).Within the GXXGXXG consensus,MtANR has the F22conserved in DFR homologs but AtANR has N at this position(Fig.2). Expression and puriﬁcation of ANR-MBP fusion protein The MtANR and AtANR open reading frames were ampliﬁed by PCR and cloned into the protein expression vector pMAL-c2X immediately downstream of the MBP to facilitate protein puriﬁcation.MBP fusion proteins have often been used for studying enzyme ki-netics,three-dimensional structures,and catalytic mechanisms[25–27].The constructs were transformed into E.coli NovaBlue(DE3).MtANR and AtANR MBP fusion proteins localized to inclusion bodies when expressed in E.coli at37°C,but were recovered as soluble enzymes following expression at16°C.This ex-pression system allowed for the production of from10 to20mg ANR protein per liter of culture,suitable amounts for crystallization studies.Fusion proteins were puriﬁed by amylose aﬃnity chromatography re-sulting in a single major band of approximatly70kDa by denaturing SDS–PAGE(Fig.3A).To ensure that the presence of the MBP did not ad-versely aﬀect the properties of ANR,we compared the speciﬁc activities and temperature optima of puriﬁed recombinant MBP-MtANR and cleaved‘‘native’’MtANR;the temperature optima were identical(45°C), and the speciﬁc activity of the native protein was re-duced by30%compared to that of the MBP fusion. Signiﬁcant protein losses occurred during separation of ANR from MBP following cleavage with Factor Xa at 4°C,although there is no Factor Xa cleavage site in the ANR protein,and this may explain the small reduction in speciﬁc activity.Eﬀects of temperature,pH,and NaCl on ANR activities The ANR reaction was linear over the30min time period used for standard assays.The temperature optima (reﬂecting the composite of reaction rate andproteinstability)of both MBP-MtANR and MBP-AtANR re-actions in vitro were quite similar (Table 1).However,dramatic diﬀerences were observed in pH dependence,with MtANR having a weakly acidic pH optimum and AtANR showing maximum activity at weakly basic pH (Figs.3B and C).The pH optimum of MtANR was 5.5in 50mM citrate/phosphate but 6.0in 50mM Mes buﬀer (Fig.3B).Addition of increasing concentrations of Na þto 50mM Mes buﬀer (pH 6.0)indicated that MtANR activity is inhibited by Na þconcentrations above200mM.This may explain the discrepancy between pH optima in citrate/phosphate and Mes buﬀer.NaCl con-centrations up to 400mM did not inhibit AtANR.Co-enzyme requirements of MtANR and AtANR Signiﬁcant diﬀerences between MtANR and AtANR were observed in their co-enzyme requirements.MtANR used either NADPH or NADH as co-sub-strates (Figs.4A and B).Double reciprocal plots of 1=VFig.3.ANR fusion protein expression in E.coli and determination of pH optima.(A)SDS–polyacrylamide (12%)gel electrophoresis of E.coli -expressed MBP-MtANR and MBP-AtANR fusion proteins stained with Commassie brilliant blue ne 1,prestained protein mass markers (New England Biolabs);lane 2,20l g crude extract from E.coli harboring pMMtANR;lane 3,20l g crude extract from E.coli harboring pMA-tANR;lane 4,2l g puriﬁed MBP-MtBAN;and lane 5,2l g puriﬁed MBP-AtBAN.(B)The eﬀect of pH on MtANR activity.(C)The eﬀect of pH on AtANR activity.In (B)and (C),curve 1is for 50mM citrate-phosphate,curve 2for 50mM Mes,and curve 3for 100mM Tris–HCl.Table 1Kinetic properties of MtANR and AtANRMtANR AtANRV max (nmol/min mg)K m (l M)k cat(min À1)k cat =K m (M À1s À1)V max (nmol/min mg)K m (l M)k cat(min À1)k cat =K m (M À1s À1)Substrates Cyanidin 1.2Æ0.712.9Æ0.410.8 1.4Â10410.0Æ1.873.9Æ5.962.7 1.4Â104Pelargonidin 14.552.8Æ3.9Delphinidin 60.649.8Æ10.880.32.7Â1040.9Æ0.0217.8Æ1.53.50.3Â104Cofactors NADPH 0.4Æ0.01450Æ120n H ¼3:3Æ0:3,½S 0:5¼125:5Æ0:7l MNADH 0.3Æ0.1940Æ155NR ÃTemperature:45°C55°C*NR,no reaction.96 D.-Y.Xie et al./Archives of Biochemistry and Biophysics 422(2004)91–102versus1/[NADPH]or1/[NADH]for MtANR indicated classic Michaelis–Menten steady-state kinetics(Figs.4A and B),with a lower K m and higher V max=K m for NADPH than for NADH(Table1).In contrast, AtANR did not use NADH as co-substrate and the saturation curve for NADPH was sigmoid rather than hyperbolic,suggesting positive cooperativity(Fig.4C).A plot of log(V=V maxÀV)versus log[NADPH]was a straight line(Fig.4D),from which a Hill co-eﬃcient (n H)of3.3and½S0:5of125l M were calculated(Table1).Properties of MtANR and AtANR towards anthocyanidin substratesMtANR and AtANR showed signiﬁcant diﬀerence in their steady-state kinetics with anthocyanidin substrates. At optimum pH,temperature,and NADPH concen-tration,MtANR exhibited Michaelis–Menten kinetics with cyanidin,pelargonidin,and delphinidin(Figs.5A–C).The double reciprocal plots indicated the highest K m for delphinidin,followed by pelargonidin and then cy-anidin(Table1).However,MtANR had its highest V max value for delphinidin(Table1),resulting in a twofold higher k cat=K m ratio for delphinidin compared to cyanidin.AtANR diﬀered markedly from MtANR in its ki-netic properties towards cyanidin and pelargonidin (Figs.5E–H),with the reaction velocity decreasing with anthocyanidin concentrations above100–150l M(Figs. 5E and G).At lower substrate concentrations,the re-action followed classic Michaelis–Menten kinetics(Figs. 5E and G).The upturn in the plots of1=V versus1/ [cyanidin]or1/[pelargonidin]on approaching the1=V axis(Figs.5F and H)is indicative of substrate inhibi-tion.This could result from low levels of a non-com-petitive inhibitor(perhaps a speciﬁc ionization product of the anthocyanidin,as depicted in Fig.1B)present in the substrate sample.The kinetics towards cyanidin and pelargonidin were thereforeﬁtted to the following ve-locity equation for a substrate plus non-competitive inhibitor:v=V max¼½S =f K mþ½S ð1þxK m=K iþx½S =K iÞg;where K m is the substrate concentration at1=2V max,x represents a coeﬃcient for[I]where½I ¼x½S in the substrate solution,and K i is the inhibitor constant de-ﬁned as f½E ½I g=½EI .This analysis facilitated calcula-tion of apparent K m values for cyanidin and pelargonidin as well as a V max for cyanidin(Table1).In contrast to the above observations with cyanidin and pelargonidin,AtANR displayed classical Micha-elis–Menten kinetics towards delphinidin(Fig.5D).The K m of AtANR was highest for cyanidin,followed by pelargonidin and then delphinidin(Table1).Neverthe-less,the k cat=K m ratio was highest with cyanidin,sug-gesting that this may be the preferred substrate in vivo.Inhibitors of MtANR and AtANRThe purity of the commercial cyanidin and pelarg-onidin substrates was checked by HPLC and was97–99%.Minor contamination byﬂavonols(kaempferol and quercetin)and dihydroﬂavonols(dihydroquercetin) was observed.(Æ)-DHQ,quercetin,and(+)-catechin (2R,3S-2,3-trans-ﬂavan-3-ol)(Fig.6A)were tested for their eﬀects on ANR activity in vitro.(+)-Catechindidnot inhibit MtANR activity(Fig.6B),but inhibited AtANR by50%at0.5mM(Fig.6C).(Æ)-DHQ did not inhibit MtANR(Fig.6D),but inhibited AtANR at concentrations as low as25l M(Fig.6E).Quercetin strongly inhibited both MtANR and AtANR(Figs.6F and G).The origin of the epimerization products of the ANR reactionWe have previously shown that ANR converts cyanidin into())-epicatechin as the major product,with ())-catechin as a minor product[12].Likewise,())-epi-afzelechin is the major product and())-afzelechin the minor product from pelargonidin.Equal amounts of ())-gallocatechin and())-epigallocatechin are formed from delphinidin[12].The question therefore arises as to whether())-catechin,())-afzelechin,and())-gallocate-chin are formed by non-enzymatic epimerization at C2 of the major products())-epicatechin,())-epiafzelechin, and())-epigallocatechin,respectively,or whether they are separate reaction products formed by the enzyme itself.To address this question,MtANR or AtANR was incubated with())-epicatechin or(Æ)-catechin in the presence of NADPH,and parallel controls were in-cluded in which active enzyme was replaced with boiled enzyme or buﬀer.No epimerization or racemization of ())-epicatechin or())-catechin into thecorresponding())-catechin or())-epicatechin,respectively,was ob-served under any of these conditions if Mes buﬀer was used.However,incubation of())-epicatechin in100mM Tris–HCl,pH7,at temperatures above30°C even without enzyme resulted in epimerization to catechin, suggesting that the())-catechin observed as a minor product in the enzyme assays might result from chemical epimerization rather than an ability of ANR to produce both2R,3R-2,3-cis-and2S,3R-2,3-trans-ﬂavan-3-ol isomers.DiscussionAmino acid sequence comparison and biochemical diﬀer-ences between MtANR and AtANRMtANR and AtANR share only60%amino acid sequence identity.Both enzymes contain the conserved Rossmann dinucleotide-binding domain at their amino-termini[24,28,29].To date,there are few available ANR gene sequences from diﬀerent plant species,and,to the best of our knowledge,no other reports of ANR sub-strate/co-substrate speciﬁcity.However,it seems likely that the two amino acid diﬀerences between the dinu-cleotide-binding domains of MtANR and AtANR(F22 and V23in MtANR corresponding to N21and L22in AtANR)may be related to the diﬀerent speciﬁcities of the two enzymes for NADPH and NADH.Examples exist where only a single amino acid change can alter co-factor binding speciﬁcity.For example,rat NADPH-dependent cytochrome P-450reductase cannot use NADH as cofactor,but mutation of Trp677to Ala generates an enzyme with quite good NADH turnover [30].Detailed structural studies of MtANR and AtANR will be necessary to deﬁnitively explain their diﬀerent co-factor speciﬁcities.The kinetics of MtANR for NADPH followed the usual Michaelis–Menten behavior observed with many reductases,whereas those for AtANR were sigmoidal and indicative of positive cooperativity,with NADPH acting as co-substrate and modulator.Otherreductases,such as ribonucleotide reductase[31],also show allo-steric behavior,and very diﬀerent kinetics of enzymes with NADPH do not necessarily require extensive dif-ferences in amino acid sequence[32,33].The non-linear kinetics of AtBAN could reﬂect the operation of the two successive hydride transfers in the overall ANR reac-tion.The turnover(k cat)of both MtANR and AtANR was quite slow.Both enzymes reduce cyanidin,pelargonidin, and delphinidin,indicating that the B-ring hydroxyl-ation pattern is not critical for catalysis.However,the relative binding aﬃnities for the three substrates diﬀer, with MtBAN exhibiting decreasing K m values in the order delphinidin>pelargonidin>cyanidin,and At-BAN exhibiting the inverse relationship.Nevertheless, both enzymes had the same k cat=K m ratio for cyanidin, which may be the major in vivo substrate for both en-zymes,consistent with the presence of())-epicatechin residues as the major building blocks in the CTs of both A.thaliana and M.truncatula.Similar to ANR,diﬀerent dihydroﬂavonol reductases(DFR)from M.truncatula, Malus domestica,and Pyrus communis showed signiﬁ-cantly diﬀerent kinetics towards DHQ and dihydroka-empferol,which diﬀer in B-ring hydroxylation pattern [34,35].High cyanidin or pelargonidin concentrations inhibit AtANR but not MtANR.This inhibition may result from the presence of non-competitive inhibitor(s)in the substrate preparations.Quercetin inhibited both MtANR and AtANR activities,whereas(+)-catechin and(Æ)-DHQ inhibited AtANR but not MtANR.M. truncatula produces(+)-catechin as a component of the seed coat CT,whereas the Arabidopsis CT does not contain(+)-catechin[15].())-Epicatechin did not in-hibit MtANR or AtANR over the tested concentration range.In the absence of information on the pool sizes of dihydroﬂavonols andﬂavan3-ols in the seed coat endothelium,it is not possible to conclude whether the above inhibitory eﬀects are of physiological signiﬁ-cance.Reaction mechanisms for reduction of anthocyanidins to ﬂavan-3-olsThe discovery of ANR explains the inversion of ste-reochemistry at C3during the biosynthesis of())-epi-catechin from leucoanthocyanidin.The heterocyclic C-ring of anthocyanidins,formed from chiral leuco-anthocyanidins,has two double bonds,at O1–C2and C3–C4,and is therefore achiral.ANR reduces anthocy-anidins toﬂavan-3-ols through NADPH-mediated re-duction at C2and C3,allowing for the inversion of stereochemistry at C3.The())-catechin(minor product) observed in enzymatic reactions may be produced by non-enzymatic epimerization at C2of())-epicatechin under the reaction conditions used.())-Catechin has recently been shown to exert strong alleopathic eﬀects and to be a major factor for colonization by the de-structive non-indigenous spotted knapweed in North America[36].Analysis of the properties of ANR from this weedy species may reveal alternative routes for the biosynthesis of())-catechin.Although anthocyanidins are often depicted as hav-ing a positive charge associated with the oxygen of the heterocyclic ring,it is recognized that the charge may be delocalized[19–21].Assessment of charge distributions on theﬂavylium cation through molecular orbital cal-culations suggests that electrophilic attack can occur at positions C8,C6and various B-ring sites,and nucleo-philic attack principally at C2and C4[16,19–21,37,38]. This is consistent with the facile in vitro chemical syn-thesis ofﬂavan-3-ols through reduction of anthocyani-dins with PtO2in a hydrogen atmosphere[38].Complete hydrogenation of cyanidin or its pentamethyl ether,and ﬁsetinidin and butinidin,resulted in the corresponding reduced products(Æ)-epicatechin and its pentamethyl ether,(Æ)-2,3-cis-3,30,40,70-tetrahydroxyﬂavan,and (Æ)-2,3-cis-30,40,7-trihydroxyﬂavan,respectively[38]. Hrazdina also synthesized(ÆÞ-30-O-methylepicatechin, (Æ)-30,50-di-O-methylepigallocatechin and(Æ)-30-O-methylepigallocatechin from peonidin(30-O-methylcy-anidin),malvidin(30,50-di-O-methyldelphinidin),and petunidin(30-O-methyldelphinidin),respectively,with sodium borohydride in ethanol or methanol[38].The intermediates of the in vitro reduction were theﬂav-3-en-3-ols andﬂav-2-en-3-ols(Fig.7),which are stable on exposure to air and in aqueous acidic solution.The initial reducedﬂavenes are in equilibrium with the cor-responding3-keto form(ﬂavan-3-one tautomer)(Fig.7), which are then further reduced to the corresponding ﬂavan-3-ols[39,40].Based on the above considerations,it is possible to postulate four possible enzymatic reduction sequences for the ANR reaction(Fig.7).In mechanism I-a,the ﬁrst hydride ion provided by NADPH/NADH nucleo-philically attacks C2to form a stableﬂav-3-en-3-ol,the conﬁguration of the C2-aryl group being2R with hydride addition by way of the b-face(addition to the a-face would be necessary to account for enzymatic production of the2S conﬁguration of the minor product ())-catechin).NADPH/NADH then provides a second hydride ion to attack C4,and a proton from the medium then quenches the charge at C3,thereby reducing the C3–C4double bond.The conﬁguration of the C3-hy-droxyl group as3R or3S is also dependent upon the direction of hydride ion addition.This will again be from the b-face to result in the formation of2R,3R-2, 3-cis-ﬂavan-3-ols.In mechanism I-b,theﬁrst step is the same as in I-a. In the second step,ANR transforms theﬂav-3-en-3-ol intermediate into its tautomericﬂavan-3-one(Fig.7) followed by reduction by NADPH/NADH at C3and100 D.-Y.Xie et al./Archives of Biochemistry and Biophysics422(2004)91–102。

甘西鼠尾草中三个新的萜类化合物

甘西鼠尾草为常用中药丹参（．ｍｉｉｒｉｇ．的同属植物，其主要成分为脂溶性二萜醌ＳｌｏｈａＢｅ）ｔｒｚ
类和水溶性酚酸类化合物 ’ ．已报道的药理活性主要有醛糖还原酶抑制活性、抑制超氧自由綦及抗氧化、保护心肌缺血及心肌缺血再灌注损伤、炎及抑菌。抗等．本课题组在对甘两鼠尾草活性部位的筛选中，证实了甘西鼠尾草对血清病性肾小球肾炎模型大鼠具有较好的治疗作用，可显著降低大鼠尿蛋白含量、减轻肾小球肿胀，还可降低正常大鼠的血液黏度，并具有显著且温和的利尿作用，以调整肾脏疾病中的电解质紊乱，对于治疗和缓解肾脏疾病的发展有积极的意可义．为了进一步考察其活性物质，本文对其活性部位的化学成分进行了深入研究，从甘西鼠尾草根和根茎的５％乙醇提取物中分离鉴定了２个新的二萜化合物和１个新的单萜苷化合物，０
１实验部分
１１试剂与仪器．
柱色谱硅胶与薄层色谱硅胶板为烟台江友硅胶开发公司产品；Ｓｐａｅ０凝胶为ＰａｍｃｅｈｄｘＩ２Ｈｈｒａｉａ
公司产品；ＤＩＯＳＣ相硅胶为Ｍｅｋ公司产品；ＩｅＣＰ０８反ｒｃＭＣｌＨ２Ｐ凝胶和ＨＰ２ｇ－０大孔吸附树脂为 ■菱
杨阳，吴志军’ ，杨颖博，来威，孙连娜，陈万生

翻译

天然产物杂志从对细胞死亡敏感和抗癌细胞系列中的海绵生物中提取抗恶性细胞增值活性的二萜异腈的评价人名和通讯地址等等暂不翻译背景信息摘要：一个新的在1位的和在已知2,4位的腈基的二萜，是从加勒比海海绵Pseudoaxinella在人类体外癌症细胞使用的线检MTT比色法检测和定量和电子扫描显微镜中分离出来的。

化合物14显示的为人类的PC3前列腺凋亡敏感的肿瘤细胞系的活动，。

化合物3和4表现出类似的增长抑制个人为三个APOP凋亡敏感和3个抗凋亡的肿瘤细胞株的定量电子显微镜分析表明，化合物1和2施加他们的活动，通过细胞毒性个人ECTS 通过抑制细胞生长的个人ECTS化合物3和4。

这些结果确定潜在的铅化合物对海洋中二萜异腈抗癌药物的发现。

萜类化合物，包含异氰和异硫氰团体往往在海洋无脊椎动物中发现的次生代谢产物，例如海绵等在过去的15年,海洋天然产物因为他们的生物学活性乎寻常的异类功能化，在科学界很多成员引起了广泛的兴趣，事实上，一个最有力的海洋抗疟药化合物是最初分离的二萜异腈，从热带海绵状物，特点是由amphilectane骨架。

在我们的研究项目的框架中, 被认为是新颖的先导化合物可以视作典型的抗癌试验，加勒比海的海绵化学海绵伪黄色生物进行了研究。

本研究引发了分离了一个二萜异腈化合物(1)和三个已知的类似的(24), 因为不同的数目和位置的异腈官能团和双键，所有这一切都是紧密的联系的。

用比色MTT检测评价生长抑菌浓度IC50(即全球经济增长的一个给定的细胞参与增殖培养了三天的化合物减少50%的浓度）的不同双萜的浓度体外测试。

定量的电子显微镜（即计算捷尔- 辅助相衬显微镜）将被用来检测是否影响细胞性通过细胞毒性或抑制细胞生长的途径。

我们已经成功地完成例如其他类型的化合物的特点包括真菌次级代谢产物, 类固醇治疗，Amar——和yllidaceae生物碱2007 Pawlik在大巴哈马海岸考察的海绵Pseudoaxinella蔺沿的（甜味剂礁）的样品被切成被冻结的小块样本然后运送到实验室用MeOH和CHCl3分别萃取提纯。

不同酶消化法提取猪原代肝细胞的效果比较

532024.4·试验研究0 引言猪圆环病毒（PCV ）是Circoviridae 科Circovirus 属的一种无囊膜的单链环状DNA 病毒。

在已知的4个血清型中，PCV2为猪易感的致病性病毒[1]。

PCV2感染会诱导宿主免疫抑制引起猪圆环病毒病（PCVD ），包括断奶仔猪多系统衰竭综合征、新生仔猪先天性脑震颤、皮炎与肾病综合征、猪呼吸道病综合征、母猪繁殖障碍等，给全世界养猪业带来较大的经济损失，是世界各国的兽医与养猪业者公认的造成重大影响的猪传染病[2]。

PCV2的感染在猪生长发育的不同阶段有不同的组织嗜性。

但无论是胎儿阶段还是出生后，肝细胞都是PCV2感染和复制的靶细胞。

因此，PCV2也被视为一种能够诱导猪肝炎的病毒[3]。

且PCV2诱导的肝细胞凋亡在PCV2引发的相关病变和疾病的发病机制中具有关键性作用[4]。

因此，方便、快捷地获取大量有活性的猪肝细胞对于研究PCVD 的致病机制具有重大意义。

目前获取肝细胞常用的方法主要包括机械分离细胞法、非酶分离细胞法、离体酶消化法和酶灌流法等[5]。

因此，本试验采用简便、经济、无需特殊设备、仅需部分肝组织的离体酶消化法，比较不同酶消化分离猪原代肝细胞的效果，为一般实验室提取分离大量有活性的猪肝细胞提供参考。

1 材料与方法1.1 材料1.1.1 主要试剂新鲜猪肝组织，Hank's 平衡盐溶液（HBSS ），磷酸盐缓冲液（无菌PBS ），4%多聚甲醛（PFA ），收稿日期：2024-01-27基金项目：国家自然科学基金项目：复杂器官与组织在脾脏内的功能性再生（32230056）作者简介：周徐倩（1999-），女，汉族，浙江温州人，硕士在读，研究方向：组织工程与再生医学。

*通信作者简介：董磊（1978-），男，汉族，安徽阜阳人，博士，教授，研究方向：组织工程与再生医学、生物材料。

周徐倩，董磊．不同酶消化法提取猪原代肝细胞的效果比较[J]．现代畜牧科技，2024，107（4）：53-55．　doi ：10.19369/ki.2095-9737.2024.04.014．　ZHOU Xuqian ，DONG Lei ．Comparison of the Eﬀect of Diﬀerent Enzyme Digestion Methods on Extraction of Porcine Primary Hepatocytes[J]．Modern Animal Husbandry Science & Technology ，2024，107（4）：53-55．不同酶消化法提取猪原代肝细胞的效果比较周徐倩，董磊*（南京大学，江苏南京 210023）摘要：猪肝细胞是猪圆环病毒的靶细胞，简单快速地提取猪原代肝细胞对于研究猪圆环病毒病的致病机制具有重要意义。

化妆品活性成分

Millicapsules A2 Millicapsules is a proprietary technology which utilizes two marine derived polymers, Alginic Acid and Agar, to produce visually distinctive skin and hair care products without the use of conventional emulsifiers. Spheres (1mm, 2mm or 4mm)are produced from these natural polymers. They can contain up to 20% emollient phase and be colored or pearlized. A2Millicapsules offer a unique alternative to conventional emulsions, serums or gels. Recommended concentration: 1-5%.INCI Name: Water (Aqua), Glycerin, Agar, Alginic Acid.Aldenine® Aldenine® is a complex of a tripeptide and hydrolyzed wheat and soy protein that boosts Collagen III synthesis while protecting cells from photo damage. Aldenine® detoxifies the skin from harmful RCS (Reactive Carbonyl Species).Recommended concentration: 2-5%.INCI Name: Hydrolyzed Wheat Protein, Hydrolyzed Soy Protein, Xanthan gum,Tripeptide-1.Alpha-Arbutin Alpha-Arbutin is a pure, water soluble, biosynthetic active ingredient. It is the more effective, faster and safer approach to promoting skin-brightening and an even skin tone on all skin types. Alpha-Arbutin also minimizes liver spots and meetsall the requirements of a modern skin-brightening and skin depigmentation product. Recommended concentration: 0.2%when formulated with an exfoliant or penetration enhancer, otherwise up to 2%.INCI Name: Alpha-Arbutin.Antarcticine® Antarcticine® is a novel high molecular weight glycoprotein obtained via biotechnology from an Antarctic strain of bacteria. Antarcticine® has cryoprotective effects providing stability to proteins and lipid membranes exposed toextremes of cold or dryness. It has also been shown to reduce the roughness of skin. Recommended concentration: 3-5%.INCI Name: Pseudoalteromonas Ferment Extract.Apt® Apt® is bio-engineered from a selected strain of red marine algae (Ahnfeltia concinna). Apt® increases cell turnover, firmness, elasticity and smoothness and brings about fine line reduction. It also contributes excellent skin feel andmoisturization properties to creams, lotions, gels and serums.INCI Name: Butylene Glycol, Ahnfeltia Concinna Extract.Argireline® Argireline® offers superior anti-wrinkle efficacy. Studies show it reduces the depth of expression wrinkles, especially in the forehead and around the eyes. It is a safer, cheaper and milder alternative to Botulinum Toxin, topically targeting thesame wrinkle-formation mechanism in a very different way. Argireline® is believed to function through modulation of theSNARE complex formulation. Recommended concentration: 3-10%.INCI Name: Acetyl Hexapeptide-8.Artemisia AO Artemisia AO, from the rare plant White Genepi, is cultivated by 100% organic methods. It is often compared to Vitamin C, and has been tested positively regarding anti-oxidative properties. It protects the skin with anti-oxidant, radicalscavenging properties, in addition to acting as an anti-inflammatory and anti-bacterial. Formulation applications includeskin-protection, anti-aging, cleansing and sun-care preparations. In addition, it is effective for reducing spotty skin.Recommended concentration: 1-3%.INCI Name: Glycerin, Water, Artemisia Umbelliformis Extract, Sodium Benzoate,Potassium Sorbate.Buddleja AO Buddleja AO, cultivated by 100% organic methods, protects the skin cells from UV damage. It has the capacity to reduce the DNA fragmentation of the keratinocytes induced by UVB radiations from 39% to 71% depending on the concentrationused. It is DNA-protective and an anti-oxidant as well. It also acts to heal skin wounds, enhancing effectiveness by itsanti-bacterial properties. Applications include anti-aging formulations, skin repair/healing and cleansing creams andlotions. Recommended concentration: 1-3%.INCI Name: Glycerin, Water, Buddleja Davidii Extract, Sodium Benzoate, PotassiumSorbate.Calendula GC Calendula GC is largely used in the care of irritated and reddened skin, against sunburn, erythema and superficial burns thanks to its anti-inflammatory, healing, soothing, hydrating, decongesting and purifying properties. Recommendedconcentration: 3-5%.INCI Name: Calendula Officinalis Flower Extract.Calendula-ECO Calendula-ECO extract is an aromatic plant of which the rich extract is produced from the flower heads. It is truly multi-functional. Due to the presence of polysaccharides, flavonoids, triterpenes and carotenes, this ECO extract is highlyeffective in tissue regeneration action. The saponins and polysaccharides are useful in moisturizing and soothingapplications. The triterpenes are also used for sensitive and irritated skin. Calendula-ECO extract also helps to preventphoto aging. Recommended concentration: 0.5-5%.INCI Name: Glycerin, Water, Calendula Officinalis Flower Extract.Cephalipin® Cephalipin® is an aqueous solution of botanically-derived lipids isolated and purified from wheat grains. These lipids help restore and normalize the natural barrier function of the stratum corneum. Cephalipin® is an ideal addition to skintreatment products. It is also compatible for after-shaving specialties, with or without ethanol. Recommendedconcentration: 2%.INCI Name: Cephalins, Disodium Cocoamphodiacetate.Ceramide I Ceramide I is a composition of C-18 Phytosphingosine base coupled to an amide-linked long-chain omega-OH fatty acid.When topically applied, Phytoceramide I optimizes dermal water retention to effect smoother, glossier and softer skin. Itis particularly suitable for treating aged or extra dry skin. This class of ceramides is believed to function as a 'molecularrivet,' cross linking lamellar structures in the barrier layer of the stratum corneum. Recommended concentration:0.01-1%.INCI Name: Ceramide 1.Ceramide III Ceramide III is a Phytosphingosine backbone acylated with stearic acid having correct 3-D human identical structure.Topical application of Ceramide III has been shown to significantly increase skin water content, reduce roughness, restoredetergent-damaged skin and protect healthy skin against detergent-induced dermatitis. Recommended concentration:0.05-1.0%.INCI Name: Ceramide 3.Ceramide IIIA Ceramide IIIA is a Phytosphingosine based human identical skin lipid in combination with the EFA, linoleic acid. Ceramide IIIA has barrier repair properties. It has additionally been shown to inhibit melanin synthesis in vitro without cytotoxicitymaking it an interesting constituent in skin lightening formulations. Recommended concentration: 0.05-1.0%.INCI Name: Ceramide 3.Ceramide IIIB Ceramide IIIB is a Phytosphingosine backbone acylated with oleic acid. Similar to Ceramide III, Ceramide IIIB differs in that it is more soluble in cosmetic emollients such as isostearic acid and isocetyl alcohol. Ceramide IIIB is recommendedin situations where its higher solubility may be required. Recommended concentration: 0.05-1.0%.INCI Name: Ceramide 3.Ceramide VI Ceramide VI is a member of the Ceramide 6 family whose chemical name is alpha-OH-stearoylphytosphingosine. It contains a long chain AHA (Alpha Hydroxyacid) found naturally in the skin which contrasts with short chain AHA's currentlyin use in skin care preparations. Efficacy studies have shown Ceramide VI to have a potent ability to smooth rough skin viamodulation of desquamation. Recommended concentration: 0.05-1.0%.INCI Name: Ceramide 6 II.Cerasol Cerasol is a 3% solution of Ceramide 3A (N-Linoleoyl-Phytosphingosine) in hexyldeconal. These ceramides are solubilized and thus bioavailable, leading to regeneration of the lipidic barrier and reduction of transepidermal water loss (TEWL). 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This activity can help delay early skin aging characterized by wrinkle formation, reduced elasticity, skin dryness and age spots. Colhibin isan ideal component for anti-aging products and well suited for sun and environmental stress protection formulations.Recommended concentration: 2-5%.INCI Name: Hydrolyzed Rice Protein.Collagen Collagen is a 1% solution of natural soluble porcine collagen. It is one of the most important scleroproteins of the connective tissue occurring as fibres. Collagen is the main structure element of the skin and is therefore largelyresponsible for its characteristic properties. It provides excellent smoothing and hydrating qualities with superb skin feel.Recommended concentration: 3-5%.INCI Name: Soluble Collagen.Complex Gel Complex Gel is a proprietary gel specially formulated to stabilize either free Vitamin A (Retinol) or Vitamin C. Complex Gel was designed to supply in dual compartment packaging with the end user freshly activating the final product at the timeof first use.INCI Name: Water, Ascorbic Acid, Glycerin, Propylene Glycol, Stearic Acid,Dimethylsilanol Hyaluronate, Hydrolyzed Wheat Protein, Carrageenans, Agar,Hydroxyethyl Cellulose, Ferrum Chloride, Zinc Chloride, EDTA, BHT.Cosmoferm® Mix III Cosmoferm® Mix III is a ready-to-use solution of Ceramide III (oil-soluble). Ceramide III and Ceramide IIIB support the renewal of the skin's natural protective layer and form an effective barrier against moisture loss. Thesehuman-skin-identical molecules are particularly suitable for long term protection and repair of sensitive and dry skin.Recommended concentration: 0.25-5.0%.INCI Name: Ceramide 3, Isocetyl Alcohol, Cetyl Alcohol.Decorinyl™Decorinyl™ is a synthesized tetrapeptide whic h mimics the sequences of decorin that bind to collagen fibrils. It has been shown to regulate fibrillogenesis and control fibril growth, improving firmness and elasticity of the skin, ensuringuniformity of fibril diameter and spacing of collagen fibrils. In effect, it replaces the non-functional decorin encountered asthe skin ages, and also sustains a superior moisture profile. Decorinyl™ has been incorporated into a liposomal system forenhanced penetration, stability and efficacy. It may be used in emulsions, gels and other formulations. Recommendedconcentration: 5%.INCI Name: Water (Aqua), Lecithin, Tipeptide-10 Citrulline, Carbomer,Triethanolamine, Phenoxyethanol, Caprylyl Glycol.DHA Film Fluid DHA (Dihydroxyacetone) Film Fluid is one of the widest used actives for sunless or self-tanning treatments. A film fluid, to stabilize DHA and enhance functionality, is created to result in a submicrodispersion, which is a stable mono-layerMolecular Film with high moisturizing effect, stabilized due to interaction with polymeric polysaccharides. The resultanttanning applications are without risks of skin cancer and aging and form a protective moisture film. DHA Film Fluid is easilyincorporated into O/W, W/O emulsions or gels. Recommended concentration: 3-5%.INCI Name: Dihydroxyacetone, Water (Aqua), Xanthan Gum, Citric Acid, SodiumHydroxide, Phenoxyethanol.Dismutin®-BT Dismutin®-BT is an aqueous solution of biotechnologically produced, highly purified superoxide dismutase from a natural yeast strain of Saccharomyces cerevisiae. It protects the skin from oxidative stress by restoring the anti-oxidant balancein the outer skin layer, thereby helping to delay early skin aging. Recommended concentration: 0.2-0.5%.INCI Name: Superoxide Dismutase.Eashave Eashave is a biopolymer complex composed of plant and biotechnologically derived materials. Eashave has proven moisturizing and soothing efficacy (supported by thermosgraphs) to skin damaged by shaving. Enhanced regeneration ofprotective lipids has also been shown.Recommended concentration: 2-5%.INCI Name: Triticum Vulgare (Wheat) Germ Extract, Saccharomyces CerevisiaeExtract, Sodium Hyaluronate.Echinacea GC Echinacia GC is a perennial, cultivated in the Swiss Alps under the highest standards of agronomic practice. A profusion of Caffeic Acid Derivatives protects skin against UV radiation damage by inhibiting degradation of Type III Collagen. ThePolysaccharide component prevents moisture loss by inhibiting Hyaluronidase enzyme actions. Essential Oils purify anddetoxify the skin and complete the smoothing, healing and firming properties of this botanical. Recommendedconcentration: 3-5%.INCI Name: Echinacea Purpura Extract.Edelweiss GC® Edelweiss GC® is a botanical extract derived from organically cultivated Edelweiss flowers grown at high altitudes in the Swiss Alps. There is environmental impact on wild strands of the rare protected plant. Plant derived glycerin is used as thecarrier which provides a well defined HPLC. Information regarding free radical scavenging, enzyme inhibitory propertiesand soothing benefits are available. Recommended concentration: 3-5%.INCI Name:Leontopodium Alpinum Extract.Edelweiss SP Edelweiss SP, an Alpine Extract, has high performing anti-oxidant and radical scavenging properties. Phenolic acids and tannins lead to increased type III collagen protection. Other compounds inhibit hyaluronidase and have anti-inflammatoryeffects, making Edelweiss SP ideal for sensitive skin in anti-aging and soothing preparations. Recommendedconcentration: 3-5%.INCI Name: Leontopodium Alpinum Extract.Elhibin® Elhibin® is a plant-derived treatment specialty which specifically inhibits skin enzymes (human leukocyte, elastase and tryptase) to minimize damage to skin from their unscheduled release. It has been clinically shown to have a soothingbenefit to skin and to improve firmness with regular use. Elhibin® normalizes the skin's enzyme/inhibitor balance. It iswell suited for anti-aging products. Recommended concentration: 3-7%.INCI Name: Glycine Soja(Soybean) Protein.Erythrulose Erythrulose, easily formulated as a high performance self-tanning product, is a natural keto-sugar, which reacts with free primary or secondary amino acid groups to form the polymer, Melanoidin. For optimal results Erythrulose is used incombination with DHA (Dihydroxyacetone). This combination is World Wide Patent Protected. It achieves a natural,long-lasting, non-streaking radiant glow, be it January or July. The skin is protected from UV Radiation, and remains moistand supple. Recommended concentrations for self-tanning products: 1-3% of Erythrulose should be combined with 2-4%DHA.INCI Name: Erythrulose.Eyeseryl® Eyeseryl® is an acetyl tetrapeptide with anti-oedema properties and a proven efficacy in reducing puffy eyebags. It also enhances skin elasticity, skin smoothness and shows a decongesting effect. Eyeseryl® can be incorporated in cosmeticformulations such as emulsions, gels, sera, etc., where a reduction of puffiness under the eye is desired. Recommendedconcentration: 1-10%.INCI Name: Water (Aqua), Acetyl Tetrapeptide-5.Fitobroside™Fitobroside™ is a lipid dispersion derived from botanical sources. It replaces key st ratum corneum lipids (glycolipids, phospholipids and sterols), helping to restore the skin's barrier function as well as providing moisturizing benefits.Recommended concentration: 3-6%.INCI Name: Triticum Vulgare (Wheat) Germ Extract.Fitoderm®: Pure, Botanically Derived Squalane... Fitoderm® is pure, botanically derived squalane, produced by the total hydrogenation of the squalene source in olive oil. It is a non-animal alternative to shark-derived squalane. Fitoderm® is highly resistant to oxidation, has a high affinity to skin and its natural lipids and is neither irritating nor allergenic. It is considered a specialty emollient and is soluble in a vast array of cosmetic solvents. Fitoderm® is perfect for moisturizing milks, creams and lotions, massage oils and a carrier for fragrance compounds.INCI Name: Squalane.Gigawhite™Gigawhite™ is a skin brightening complex of botanical origin, which was developed with the help of 'in-vitro' assays, human cell cultures and a skin model with barrier effect. Gigawhite™ at 5% has been proven to have a significant skinbrightening effect with regular topical use. It was additionally shown to reduce the color and size of age spots in a twelveweek 'in-vivo' study. Gigawhite™ is appropriate in topical emulsions, gels and aserums. Recommended concentration:3-5%.INCI Name: Malva Sylvestris (Mallow) Extract, Mentha Piperita (Peppermint) LeafExtract, Primula Veris Extract, Alchemilla Vulgaris Extract, Veronica OfficinalisExtract, Melissa Officinalis Leaf Extract, Achillea Millefolium Extract.Ginger-ECO Ginger-ECO, one of the best known botanicals since ancient times, has excellent anti-aging properties due to its ability to almost totally inhibit collagen degradation by collagenase enzyme inhibition. It allows the maintenance of protein levels inthe dermis and skin flexibility. It also acts as an anti-oxidant and anti-inflammatory. Ginger-ECO is used in formulationsfor skin and hair products, including refirming, massage and anti-dandruff preparations. Recommended concentration is0.1-2%.INCI Name: Glycerin, Water, Zingiber Officinate (Ginger) Root Extract.Ginkgo-ECO Ginkgo-ECO extract is produced from the leaves of the Ginkgo Biloba tree, and is multi-functional. The basis for its cosmetic benefits are two groups of compounds: Flavonoids and terpenes. The flavonoids indicate several properties:Antioxidant, anti-inflammatory, vessel protection, lipolysis stimulation and cell regeneration. Ginkgolides, the terpenes,have been demonstated to reduce platelet aggregation, allergic reactions and general inflammatory responses due to theirPAF antagonistic activity. These two groups promote anti-cellulite, anti-aging, photo-protection, re-epithelizing and othercosmetic advantages in formulation. Recommended concentration: 0.1-3%.INCI Name: Glycerin, Water, Ginkgo Biloba Leaf Extract.Glucare™ S 2%Glucare™ S 2% is an aqueous solution containing 2% of carboxymethylated derivative of 1, 3-Beta Glucans from yeast.The degree of carboxymethylation is carefully controlled to achieve good solubility while maintaining optimal biologicalactivity. Studies demonstrating soothing and immunoprotective effects are available. Glucare™ S 2% also has anexcellent silky skin feel. Recommended concentration: 2.5-10%.INCI Name: Sodium Carboxymethyl Betaglucan.Glucare™ S Powder Glucare™ S Powder is a form of Glucare™ S (2%). The degree of carboxymethylation is carefully controlled to achieve good solubility with optimal biological activity. Studies demonstrating soothing and immunoprotective effects areavailable. Recommended concentration: 0.05-0.40%.INCI Name:Sodium Carboxymethyl Betaglucan.Green Tea-ECO Green Tea-ECO is an active of diverse advantages, primarily due to the content of Polyphenols and Methylxanthines. The Polyphenols are responsible for the anti-oxidant and anti-inflammatory effects. In addition, they have the ability to inhibitcollagenase, the enzyme causing degradation of collagen and the vascular endothelium. The Methylxanthines promotevasodilator response, and prevent the lipid accumulation into the adipocytes. Green Tea-ECO also protects against UVBdamage. Applications include anti-cellulite treatments, sun-protectors, anti-wrinkling formulations, irritated and sensitiveskin products, and hair care products that improve strength and gloss. Recommended concentration: 0.1-2%.INCI Name: Camellia Sinensis.Happy Skin Happy Skin is a novel, natural active ingredient, derived from the Roseroot plant. It demonstrates the important dynamic interactions between the skin and the nervous system. Because of the stimulation of neuro-peptide production, and its antioxidantproperties, Happy Skin promotes inner wellness, relaxation and stress reduction that becomes manifest in the external appearanceof the body and facial skin and hair. Recommended concentration: 0.2-2%.INCI Name: Glycerin, Water (Aqua), Rhodiola Rosea Root Extract.Hibiscus-ECO The flowers of this plant have anti-oxidant properties and inhibit elastin degradation, resulting in maintaining the skin's elasticity and consequent anti-aging effects. It is useful for formulations targeting skin tone, anti-cellulite and refirmingproducts. Recommended concentration: 0.1-4%.INCI Name: Hibiscus Sabdariffa.Hispagel® 100 Hispagel® 100 is a hydrated polymeric clathrate containing vegetally-derived glycerin that is readily soluble in water, forming permeable films on the skin with excellent moisture retention and lubricating properties. It has excellentthickening and stabilizing properties for use in all types of personal care products.INCI Name: Glycerin, Glyceryl Polyacrylate.Hispagel® 200 Hispagel® 200 is a hydrated polymeric clathrate containing vegetally-derived glycerin that is readily soluble in water, forming permeable films on the skin with excellent moisture retention and lubricating properties. Excellent thickening andstabilizing properties for use in all types of personal care products.INCI Name: Glycerin, Glyceryl Polyacrylate.Hispagel® Oil LV Hispagel® Oil LV is a new, low viscosity hydrogel developed for topical use with an excellent skin feel and moisturizing properties. Hispagel® Oil is an oil-free hydrophilic system that delivers lubricity and emolliency in topical formulations.The LV form has a viscosity range of 500-1200 cps.INCI Name: Glycerin, Glyceryl Polyacrylate.Homeostatine® Homeostatine® is an active complex of two natural botanical ingredients, which through an innovative process, prevent and reduce wrinkles by recovering and maintaining homeostasis in the dermal extra-cellular matrix. Homeostatine®increases the production of dermal collagen and other ECM components in fibroblasts. It inhibits the synthesis ofMetalloproteinases (MMP) as well as the synthesis of pro-inflammatory mediators. The result is less wrinkled, more elasticand firmer skin. Recommended concentrations: 2-3% for early wrinkles and 2-5% for aged skin.INCI Name:Enteromorpha Compressa Extract, Caesalpinia Spinosa Gum.Houseleek-ECO Houseleek-ECO is an extract produced from the aerial part of Houseleek that contains multi-functional benefits for cosmetic uses. Astringent activity is due to the tannin content-regulating sebum secretions and promoting tissueregeneration, especially on irritated skin. Antioxidant and lipid level-decreasing properties help guard against damagingoxidative processes to skin and hair. Antimicrobial activity of phenols in Houseleek inhibits growth of microorganisms,enabling formulations for purifying and antiseptic products. The mucilage and organic acids found in the extract regulateTEWL and are especially gentle for sensitive skin. Recommended concentration: 0.1-3%.INCI Name: Glycerin, Water, Sempervivum Tectorum Extract.HSC Hydrophobic Sphingolipid Complex (HSC) contains ceramides, incorporated into an organo-compatible silicone/emollientmatrix. This matrix includes cyclomethicone, dimethiconol, phenyl trimethicone, octyl cocoate and phytosterols. HSC helps restore, replenish and restructure lipids within the stratum corneum. Extraordinary skin feel. Recommended concentration: 5-10%.INCI Name: Cyclopentasiloxane, Cyclotetrasiloxane, Dimethiconol, Dimethicone, Ethtylhexyl Cocoate, Phenyl Trimethicone, Lecithin, Glycolipids.Hyaluronic Acid Powder-BT Hyaluronic Acid Powder-BT has water storing properties and is an ideal swelling agent and lubricant, enabling its incorporation into cosmetics leading to a perceptible and visible improvement of skin condition. It forms a thin transparent visco elastic surface film. The film helps to preserve characteristics of youthful and healthy skin, suppleness, elasticity and tone. Recommended concentration: 0.01-1%.INCI Name: Sodium Hyaluronate.Hyaluronic Acid Solution-BT (Hyasol®-BT) Hyaluronic Acid Solution-BT (Hyasol®-BT) is a 1% solution of Hyaluronic Acid-BT. This naturally-derived (biofermentation) material is produced with special attention to maintaining its high relative molecular weight. Its excellent purity is assured though an innovative biofermentation process. Recommended concentration: 3-5%. INCI Name: Sodium Hyaluronate.Hydrolastan® Hydrolastan® is a natural scleroprotein and aqueous solution containing 10% of high molecular weight elastin hydrolysate. With high affinity to skin and hair, Hydrolastan® provides a natural protein layer of protection againstenvironmental factors. It has excellent smoothing properties and substantivity to skin. Recommended concentration:5-10%.INCI Name: Hydrolyzed Elastin.Hydromanil Hydromanil is a unique, three dimensional glycol-matrix delivery system that releases moisturizing molecules sequentially into the stratum corneum, resulting in a highly significant improvement of both immediate and long term skinhydration. Solubilized in water and propylene glycol. Recommended concentration: 2-10%.INCI Name: Propylene Glycol, Glycerin, Caesalpinia Spinosa Oligosaccharides,Caesalpinia Spinosa Gum (Pending).Hydromanil H.GL Hydromanil H.GL is a unique, three dimensional glycol-matrix delivery system that releases moisturizing molecules sequentially into the stratum corneum, resulting in a highly significant improvement of both immediate and long term skinhydration. Solubilized in water and glycerin. Recommended concentration: 2-10%.INCI Name: Glycerin, Hydrolyzed Caesalpinia Spinosa Gum, Caesalpinia SpinosaGum (Pending).Hypericum PG Hypericum PG extract is widely used for irritated and reddened skin, against sunburn, erythema and superficial burns.Hypericum PG is therefore well suited for after-sun agents as well as for healing products and acne-prone skin.Recommended concentration: 3-5%.INCI Name: Hypericum Perforatum Extract.Hyssopus AO The Hyssop plant is organically cultivated in the Alps for its diverse skincare and hair care properties. Its special acids, tannins, flavenoids and essential oils give this ingredient a range of functionalities. It may be used for products targetingacne prone skin, cleaning formulations requiring anti-bacterial properties, and anti-oxidant skincare protection, as well asrinse-off bath/shower and shampoo formulations. Recommended concentration: 1-3%.INCI Name: Glycerin, Water, Hyssopus Officinalis Extract, Sodium Benzoate, PotassiumSorbate.Immucell® Immucell® is a biotechnologically-derived fraction which is rich in oligopeptides. Immucell® is highly stimulatory to cell respiration. It has been shown to boost the immune system of skin and increase cell turnover. Recommendedconcentration: 2-6%.INCI Name: Glycoproteins.Imperatoria AO Imperatoria AO is cultivated by Alpaflor using 100% organic methods in the Alps. The Masterwort plant extract is composed of beneficial flavenoids, phenolic acids, tannins, sugars and essential oils. Its properties includere-epithelialization, anti-inflammatory functions, moisturization, antioxidant radical scavenging and astringency. It can beapplied in formulations for after-sun preparations, stretch mark treatments and skin repair products. Recommendedconcentration is 1-3%.INCI Name: Glycerin, Water, Peucedanum Osthruthium Leaf Extract, Sodium Benzoate,Potassium Sorbate.Ion-Moist 4Men Ion-Moist 4Men is a molecular film, specifically formulated for men, that provides natural hydration due to its NMF ingredient content (amino acids, sugars, lactate, urea, PCA and several ions). Ion-Moist 4Men restores the levels ofmoisture after shaving and preserves the natural barrier function of the epidermis. It stimulates moisture retention in thestratum corneum and enhances smoothness of the skin. Men tend to have more collagen than women, but less hydration.This unique ingredient can be used in O/W emulsions, body milks, sera, aftershave gels and other preparations. At least3-10% of the ingredient should be added to the final formulation.INCI Name: Water (Aqua), Glycerin, Calcium Pantothenate, Xanthan Gum, Urea,Caprylyl Glycol, Magnesium Chloride, Postassium Chloride, Postassium Lactate,Magnesium Lactate, Sodium Citrate, Glucose, Citric Acid, Ethylhexylglycerin.Iricalmin Iricalmin is a plant-derived, multi-functional biopolymer complex for use in skin care products. Iricalmin helps regenerate the protective lipid layer of damaged skin while delivering moisturization. It has also been shown to have soothingbenefits. Iricalmin is suitable for after-sun, after-depilation and after-shave products. Recommended concentration:2-5%.INCI Name: Triticum Vulgare (Wheat) Germ Extract, Saccharomyces CerevisiaeExtract, Sodium Hyaluronate.Lactomide Lactomide is an aqueous liposomal dispersion containing a naturally derived ceramide (ceramide-3) as well as phosphatidylethanolamine and phosphatidylcholine (milk lipids). The regenerating and moisture-regulating effect ofLactomide is due to the fact that its active substances are central components of the epidermal permeability barrier.Recommended concentration: 2-5%.INCI Name: Milk Lipids, Ceramide 3.Legactif Legactif is a complex of plant active ingredients. It is rich in flavonoids, such as rutin, hesperidin and naringin. Steroidal saponins are also major components for personal care applications. Legactif aims at relieving 'tired leg syndrome' and legheaviness, which affects over 1/2 of the adult population. Its cosmetic properties include blood circulation stimulation and。

Salinomycin_53003-10-4_DataSheet_MedChemExpress

P d D Sh Product Name:Salinomycin CAS No.:53003-10-4Cat. No.:HY-15597Product Data SheetMWt:751.00Formula:C42H70O11Purity :>98%Solubility:DMSOMechanisms:Biological Activity:Salinomycin is an antibacterial and coccidiostat ionophore therapeutic drug.Pathways:Anti-infection; Target:Antibacterial y p p gIC50 Value:Target: Antibacterial; Anticancer Salinomycin, a polyether ionophore antibiotic isolated from Streptomyces albus, has been shown to kill CSCs in different types of human cancers, most likely by interfering with ABC drug transporters,the Wnt/β-catenin signaling pathway, and other CSC pathways[1].in vitro: Salinomycin acts as a potassium ionophore and thereby interferes with transmembrane potassium potential, leading to mitochondrial and cellular potassium efflux. low concentrations of Salinomycin of 1 μM and 2 μM resulted in haggard and less confluent grown cells, increasing t ti f S li i f 5M d 10M lt d i l b l d d f t d ll l References:[1]. Naujokat C, Steinhart R. Salinomycin as a drug for targeting human cancer stem cells. J BiomedBiotechnol. 2012;2012:950658.[2]Thorsten Lieke Wolf Ramackers Sabine Bergmann Impact of Salinomycin on human concentrations of Salinomycin of 5 μM and 10 μM resulted in a globular and defragmented cellular phenotype[2]. Treatment of 10.2 uM Sal for 48 h caused G0/G1 cell cycle arrest in HepG2 cells,whereas 15.3 uM...[2]. Thorsten Lieke, Wolf Ramackers, Sabine Bergmann, Impact of Salinomycin on humancholangiocarcinoma: induction of apoptosis and impairment of tumor cell proliferation in vitro. BMCCancer. 2012; 12: 466.[3]. Fan Wang, Lei He, Wei-Qi Dai, Salinomycin Inhibits Proliferation and Induces Apoptosis ofHuman Hepatocellular Carcinoma Cells In Vitro and In Vivo. PLoS One. 2012; 7(12): e50638.Caution: Not fully tested. For research purposes onlyMedchemexpress LLC18 W i l k i n s o n W a y , P r i n c e t o n , N J 08540,U S AE m a i l : i n f o @m e d c h e m e x p r e s s .c o m W e b : w w w .m e d c h e m e x p r e s s .c om。

乳鼠原代心肌细胞的英语

乳鼠原代心肌细胞的英语英文回答：Neonatal Rat Primary Cardiomyocytes Isolation and Culture.Neonatal rat primary cardiomyocytes (NRCMs) areisolated from the hearts of newborn rats and cultured in vitro as a model system for studying cardiac biology and function. These cells are highly differentiated and exhibit many of the characteristics of adult cardiomyocytes, including the ability to contract spontaneously and respond to pharmacological agents. NRCMs have been used extensively in research to investigate a variety of cardiovascular diseases, including heart failure, arrhythmias, and ischemic injury.Isolation of NRCMs.NRCMs are typically isolated from 1to 3-day-oldSprague-Dawley rats. The rats are euthanized and the hearts are removed and placed in sterile phosphate-buffered saline (PBS). The hearts are then minced into small pieces and digested with a collagenase solution. The resulting cell suspension is filtered and centrifuged to separate the cardiomyocytes from other cell types.Culture of NRCMs.The isolated NRCMs are resuspended in a culture medium supplemented with serum and antibiotics and plated onto culture dishes. The cells are allowed to adhere to the dishes for 24 hours, after which the medium is replaced with a serum-free medium. NRCMs can be cultured for up to 4 weeks, although they typically begin to lose their differentiated characteristics after 2-3 weeks.Characterization of NRCMs.NRCMs can be characterized by their morphology, electrophysiological properties, and contractile function. Morphologically, NRCMs are polygonal in shape and have acentral nucleus. They exhibit spontaneous contractions and respond to electrical stimulation. NRCMs express a varietyof cardiac-specific proteins, including sarcomeric proteins, ion channels, and calcium-handling proteins.Applications of NRCMs.NRCMs have been used in a wide range of research applications, including:Investigation of cardiac diseases: NRCMs have beenused to study the mechanisms underlying a variety of cardiovascular diseases, including heart failure, arrhythmias, and ischemic injury.Development of new drugs: NRCMs have been used to screen for new drugs that may be effective in treating cardiovascular diseases.Toxicology testing: NRCMs have been used to test the toxicity of new drugs and chemicals.Gene therapy: NRCMs have been used to study the effects of gene therapy on cardiac function.Advantages of NRCMs.NRCMs offer a number of advantages over other cell types for studying cardiac biology and function. These advantages include:High degree of differentiation: NRCMs are highly differentiated and exhibit many of the characteristics of adult cardiomyocytes.Spontaneous contractility: NRCMs exhibit spontaneous contractions, which makes it possible to study cardiac function without the use of electrical stimulation.Response to pharmacological agents: NRCMs respond to pharmacological agents in a manner similar to adult cardiomyocytes, which makes them a good model system for studying the effects of drugs on cardiac function.Easy to isolate and culture: NRCMs are relatively easy to isolate and culture, which makes them a cost-effective and convenient model system.Disadvantages of NRCMs.NRCMs also have some disadvantages, including:Limited lifespan: NRCMs can only be cultured for up to 4 weeks, which limits their use for long-term studies.Loss of differentiated characteristics: NRCMs begin to lose their differentiated characteristics after 2-3 weeks in culture, which limits their use for studying chronic cardiac diseases.Variability between preparations: The isolation and culture conditions can affect the properties of NRCMs, which can lead to variability between preparations.中文回答：新生大鼠原代心肌细胞——分离与培养。

New Anti-inflammatory 4-Hydroxyisoflavans from Solanum lyratum

The whole plant of Solanum lyratum T HUNB(Solanaceae) is a renowned traditional Chinese medicine that has been used as anti-inﬂammatory, antitumor, antibacterial, and an-tioxidant agent.1—4)In our previous phytochemical studies, we reported isolation of several sesquiterpenoids, ﬂavonoids, and amides, some of which showed signiﬁcant cytotoxic ac-tivities.5—8)As part of our ongoing search for new natural compounds with biological activities, the ethanolic extract of S. lyratum was further investigated, which led to the isolation of three new 4-hydroxyisoﬂavans, named lyratin A (1), lyratin B (2), and lyratin C (3), together with a known 4-hydroxyisoﬂavan,4,7,2Ј-trihydroxy-4Ј-methoxyisoﬂavan (4). On the basis of extensive spectroscopic analyses, the structures of the new compounds were elucidated. In addi-tion, the isolated four 4-hydroxyisoﬂavans were screened for their anti-inﬂammatory activities with inhibitory rates of re-lease of b-glucuronidase from polymorphonuclear leuko-cytes of rats in the range of 30.3—38.6% at 10m M. H erein we report the isolation, structure elucidation and anti-inﬂam-matory activities of these four 4-hydroxyisoﬂavans. Results and DiscussionCompound 1was isolated as a yellow gum. The molecularformula was determined C20H22O5by H R-electrospray ion-ization (ESI)-MS, which indicated a quasi-molecular ion peak at m/z343.1541 [MϩH]ϩ. The IR spectrum showed ab-sorption bands at 3410, 1605, 1558, and 1459cmϪ1, which were in agreement with hydroxy and aromatic groups. The 1H-NMR spectrum showed the presence of a pair of doubletsat dH 3.57 (1H, dd, Jϭ11.0, 11.1H z) and 4.19 (1H, dd,Jϭ5.0, 11.0Hz), a multiplet at dH 3.52 (1H, m), and a dou-blet at dH 5.45 (1H, d, Jϭ6.6Hz). These signals were assign-able to two H-2 protons, H-3, and H-4 protons of a 4-hydrox-yisoﬂavan skeleton. The corresponding carbons were identi-ﬁed by heteronuclear multiple quantum coherence (HMQC)experiment as a methylene carbon at dC 65.8 (C-2) and twomethine carbon atoms at dC 39.3 (C-3) and 77.4 (C-4). In the1H-NMR spectrum, further signals were observed thatshowed the presence of 7-substituted ring A [dH 7.25 (1H, d,Jϭ8.4Hz, H-5), 6.48 (1H, dd, Jϭ2.0, 8.4Hz, H-6), and 6.25 (1H, d, Jϭ2.0H z, H-8)], ortho-coupled aromatic doublets[dH 6.32 (1H, d, Jϭ8.0Hz, H-5Ј) and 6.92 (1H, d, Jϭ8.0Hz,H-6Ј)], a prenyl unit [dH 3.12 (2H, d, Jϭ6.9Hz, H-1Љ), 5.15(1H, t, Jϭ6.9Hz, H-2Љ), 1.67 (3H, s, H-4Љ), and 1.59 (3H, s,H-5Љ)], and three hydroxyl units [dH 9.59 (1H, s), 9.21 (1H,s) and 8.31 (1H, s)]. In the heteronuclear multiple bond cor-relation (HMBC) spectrum, the cross peaks from the protonat dH9.59 to C-7, from the proton at dH9.21 to C-4Ј, andfrom the proton at dH8.31 to C-2Јconﬁrmed that three hy-droxyl moieties were attached to C-7, C-4Ј, and C-2Ј, respec-tively. The long-range correlations from H-4 to C-2, C-3, C-5, C-9, and C-1Ј, from H-3 to C-2, C-10, C-1Ј, C-2Ј, and C-6Ј, as well as from H-2 to C-3, C-4, and C-1Јindicated thepresence of the C-ring. The position of the prenyl group wasconﬁrmed by HMBC experiment, which showed correlationbetween a vinylic proton H-2Љof the prenyl substituent andC-3Ј, furthermore, the methylene protons H-1Љof the prenylsubstituent showed correlations with C-2Ј, C-3Ј, and C-4Ј.The proton H-6Јshowed correlations with C-2Ј, C-3Ј, C-4Ј,and C-3, while proton H-5Јshowed correlations with C-1Ј,C-3Ј, and C-4Ј. Thus the substitution in the B-ring was deter-mined to be 2Ј,4Ј-dihydroxy-3Ј-g,g-dimethylallyl. The rela-tive conﬁguration of 1was determined from rotating frameOverhauser enhancement spectroscopy (ROESY) data. In theROESY spectrum, cross peaks were observed from H-3 toH-2band from H-4 to H-2a, conﬁrming that the relative con-ﬁguration of H-3 and H-4 was trans. Furthermore, the ab-solute conﬁgurations of C-3 and C-4 of 1were establishedfrom circular dichroism (CD) spectrum. In a previouspaper,9)it was reported that (3S,4R)-4-hydroxyisoﬂavan ex-hibits a negative Cotton effect at 220—250nm and a positiveCotton effect at 250—300nm, while (3R,4S)-4-hydroxy-isoﬂavan showed positive and negative Cotton effects in theregions of 220—250 and 250—300nm, respectively. Sincethe CD spectrum of 1showed a negative Cotton effect at239nm and a positive Cotton effect at 285nm, this suggeststhe absolute conﬁgurations of C-3 and C-4 as 3S, and 4R, re-spectively. On the basis of the above data and comprehensive2D NMR experiments (HMQC and HMBC), the structure ofcompound 1was identiﬁed as shown in Fig. 1, named lyratinA.Compound 2was obtained as a brown paste and gave aHR-ESI-MS ion peak at m/z341.1385 [MϩH]ϩ, correspond-ing to a molecular formula of C20H20O5. The IR spectrumdisplayed absorption bands at 3431, 1610, 1568, and 1457cmϪ1, which were assignable to hydroxy and aromatic moi-eties. Comparison of its 1H-NMR spectrum with that of 1showed that 2had many features in common with 1. In con-trast to compound 1, the 1H-NMR spectrum of 2displayedsignals of one 2,2-dimethylpyran moiety [dH5.68 (1H, d,840V ol. 58, No. 6 NoteNew Anti-inﬂammatory 4-Hydroxyisoﬂavans from Solanum lyratumDe-Wu Z HANG,a Gui-Hai L I,b Qun-Ying Y U,a and Sheng-Jun D AI*,aa School of Pharmace utical Scie nce, Yantai Unive rsity; Yantai 264005, Pe ople’s Re public of China: andb ShandongAcademy of Chinese Medicine; Jinan 250014, People’s Republic of China.Received November 21, 2009; accepted February 1, 2010Three new 4-hydroxyisoﬂava ns, na med lyra tin A (1), lyra tin B (2) a nd lyra tin C (3), a long with a known compound, 4,7,2؅trihydroxy-4؅-methoxyisoﬂa va n (4), were isola ted from the whole pla nt of Solanum lyratum.Their structures were established by means of detailed physical data analyses. In vitro, four compounds showedanti-inﬂammatory activities with inhibitory ratios of release of b-glucuronidase from polymorphonuclear leuko-cytes of rats in the range of 30.3—38.6% at 10m M.Key words Solanum lyratum; Solanaceae; 4-hydroxyisoﬂavan; lyratin; anti-inﬂammatory activityChem. Pharm. Bull.58(6) 840—842 (2010)© 2010 Pharmaceutical Society of Japan ∗To whom correspondence should be addressed.e-mail: daishengjun_9@J ϭ9.9Hz, H-3Љ), 6.37 (1H, d, J ϭ9.9Hz, H-4Љ), 1.35 (3H, s,H-5Љ), and 1.33 (3H, s, H-6Љ)], which was cyclized with C-4Јand C-2Љby an ether linkage, as supported by corresponding HMBC experiment. The relative conﬁguration of the stereo-genic centers of 2was the same as those in 1, as determined by the ROESY spectrum. Furthermore, based on the results of optical rotation and CD data, the absolute conﬁgurations of two chiral carbons of 2were found in agreement with 1.Consequently, the structure of compound 2was established as shown in Fig. 1, named lyratin B.Compound 3was isolated and puriﬁed as a pale yellow gum, and the molecular formula was established as C 20H 22O 6by H R-ESI-MS, which displayed a quasi-molecular ion at m /z 359.1490 [M ϩH]ϩ. The IR spectrum exhibited absorp-tion bands at 3433, 1611, 1552, and 1455cm Ϫ1, which corre-sponded to hydroxy and aromatic groups. The 1H- and 13C-NMR spectra of 3were similar to those of 2except for the absence of the double bond across C-3Љ/C-4Љand the pres-ence of an oxygenated methine proton [d H 3.58 (1H , dd,J ϭ5.3, 7.4Hz, H-3Љ); d C 67.4], a methylene moiety [d H 2.77(1H , dd, J ϭ5.3, 16.8H z, H a -4Љ) and 2.33 (1H , dd, J ϭ7.4,16.8H z, H b -4Љ); d C 25.9], and a hydroxyl group [d H 5.09(1H, d, J ϭ4.7Hz)]. The long-range correlation between the proton at d H 5.09 and C-3Љindicated that the hydroxy group was attached to C-3Љ. With the aid of the ROESY data, it was readily conﬁrmed that the relative conﬁguration of the pro-tons at C-3 and C-4 was also trans . Additionally, the absolute conﬁgurations of C-3 and C-4 were assigned as 3S , 4R on thebasis of its CD spectrum, which displayed a negative Cotton effect at 239nm and a positive Cotton effect at 285nm.9)In this way, except for C-3Љ, the absolute conﬁgurations of all chiral centers of 3were established. Thus the structure of compound 3was determined as shown in Fig. 1, named lyratin C.The known compound was identiﬁed as 4,7,2Ј-trihydroxy-4Ј-methoxyisoﬂavan (4) by comparison of its physical and spectral data with those reported in the literature.10)Compounds 1—4and ginkgolide B were evaluated for anti-inﬂammatory activities using established methods,11,12)and the inhibitory rates of release of b -glucuronidase from polymorphonuclear leukocytes (PMNs) of rats were 31.7%,38.6%, 30.3%, 35.4% and 50.2% respectively at 10m M .Based on the bioassay results, it may be concluded that these compounds have inhibitory activities on the release of b -glu-curonidase from rat PMNs induced by platelet activating-fac-tor (PAF).ExperimentalGeneral Experimental Procedures Optical rotations were recorded by Perkin-Elmer 241 polarimeter. UV spectra were obtained by Shimadzu UV -160 spectrophotometer. IR spectra were determined by Perkin-Elmer 683 in-frared spectrometer with KBr disks. CD spectra were recorded by JASCO-815 CD spectrometer. ESI-MS were measured by Bruker Esquire 3000 Plus spectrometer. HR-ESI-MS were recorded by Micromess Q-Tof Global mass spectrometer. NMR spectra were obtained by Varian Unity BRUKER 400 at 400MHz (1H) and 100MHz (13C) with tetramethylsilane (TMS) as internal standard. Silica gel (200—300 mesh) for column chromatography and silica gel GF254 for preparative TLC were obtained from Qingdao Marine Chemi-cal Factory, Qingdao, People’s Republic of China.Plant Material Solanum lyratum T HUNB was collected in Linyi district,Shandong Province, People’s Republic of China, in September 2006, and identiﬁed by Professor Y an-yan Zhao, School of Pharmaceutical Science,Y antai University. The whole plant of S. lyratum was harvested and air-dried at room temperature in the dark. A voucher specimen (YP06089) has been deposited at the herbarium of the School of Pharmaceutical Science, Y antai University.Extra ction a nd Isola tion The air-dried whole plant of S. lyratum (20.0kg) was ﬁnely cut and extracted three times (1h ϫ3) with reﬂuxing EtOH . Evaporation of the solvent under reduced pressure provided the ethanolic extract. The extract was dissolved and suspended in H 2O, and par-titioned with CHCl 3, EtOAc, and n -BuOH. The CHCl 3fraction (217.1g) was initially subjected to silica gel column (10ϫ90cm) chromatography (200—300 mesh, 2.0kg) and eluted with cyclohexane–acetone at 95:5 (6.0l),90:10 (6.0l), 85:15 (6.0l), 80:20 (7.0l), 75:25 (7.0l), 70:30 (7.0l),60:40 (5.0l), and 50:50 (3.0l) to give ten fractions. Fraction 5 (3.6g) was separated by CC over silica gel [eluted by cyclohexane–acetone, (100:0—70:30)], Sephadex LH -20 [100g, eluted with EtOAc–EtOH , 50:50, v/v],and preparative TLC [CH Cl 3–EtOAc, 7:1, v/v] to afford compounds 2(18.6mg) and 4(11.3mg). Fraction 7 (4.2g) was isolated by CC on silica gel [eluted by CHCl 3–CH 3COCH 3, (100:0—60:40)] and preparative TLC [CHCl 3–EtOAc, 4:1, v/v] to give compounds 1(25.4mg) and 3(39.1mg).Lyratin A (1): A yellow gum. [a ]D 25Ϫ113.5°(c ϭ0.36, CH Cl 3). UV (CHCl 3) l max : 209 and 297nm. IR (KBr) n max : 3410, 1632, 1605, 1558,1459 and 1023cm Ϫ1. CD (c ϭ0.87ϫ10Ϫ3, MeOH ) D e (nm): Ϫ1.37 (239),ϩ1.40 (285). ESI-MS m /z : 343.3 [M ϩH]ϩ. H R-ESI-MS m /z : 343.1541[M ϩH]ϩ(Calcd for C 20H 23O 5, 343.1545). 1H - and 13C-NMR data, see Table 1.June 2010841Fig.1.The Structures of Compounds 1—4Isolated from Solanum lyra-tumFig.2.Key HMBC Correlations of Compounds 1—3Fig.3.Key ROESY Correlations of Compounds 1and 2842V ol. 58, No. 6。

疣孢菌FIM090395细胞毒活性次级代谢产物的研究

疣孢菌FIM090395细胞毒活性次级代谢产物的研究吴云丹;聂毅磊;陈名洪;林如;谢阳;方东升;彭飞;连云阳;江红【摘要】目的研究分离自海洋沉积物的疣孢菌FIM090395产生的次级代谢产物.方法采用HP-20大孔吸附树脂、正相硅胶、Sephadex LH-20葡聚糖凝胶和反相C18柱层析等方法,对菌株FIM090395次级代谢产物进行分离纯化.通过紫外光谱、质谱、核磁共振等波谱学方法对化合物进行结构鉴定.结果与结论分离得到化合物1和化合物2,结构解析确定它们分别与proximicin A和B同质,活性研究表明化合物1对肿瘤细胞株MDA-MB-231(乳腺癌细胞)、SW620(结肠癌细胞)、SMMC7721(肝癌细胞)和CNE-2(鼻咽癌细胞)具有一定的抑制作用,其IC5o值分别为23.49、13.48、34.96和16.28μg/mL;化合物2对肿瘤细胞株K562(白血病细胞)具有一定的抑制作用,其IC50值为25.69μg/mL,对MDA-MB-231和CNE-2的抑制增殖作用较弱.【期刊名称】《中国抗生素杂志》【年(卷),期】2014(039)002【总页数】4页(P102-105)【关键词】疣孢菌;肿瘤细胞毒活性;次级代谢产物;Proximicins【作者】吴云丹;聂毅磊;陈名洪;林如;谢阳;方东升;彭飞;连云阳;江红【作者单位】福建医科大学药学院,福州350108;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007;福建医科大学药学院,福州350108;福建省微生物研究所福建省新药(微生物)筛选重点实验室,福州350007【正文语种】中文【中图分类】R979.1+4近年来，不断有来自海洋微生物的新种、属的报道[1]，2010年Goodfellow等[2]统计了目前在海洋环境中发现的放线菌属有50个，其中新属有12个。

儿童急性髓系白血病的遗传基因异常及其意义

中国小儿血液与肿瘤杂志２０２１年６月第２６卷第３期ＪＣｈｉｎａＰｅｄｉａｔｒＢｌｏｏｄＣａｎｃｅｒ，Ｊｕｎｅ２０２１，Ｖｏｌ２６，Ｎｏ．３
·１３１·
２异常基因在ＭＲＤ监测中的作用２１融合基因ＲＵＮＸ１ＲＵＮＸ１Ｔ１／ｔ（８；２１）和ＣＢＦＢＭＹＨ１１／ｉｎｖ（１６）／ｔ（１６；１６）均可用于治疗后的ＭＲＤ监测。［１，８１１］诱导或巩固结束后，ＲＴｑＰＣＲ检测其转录产物ｍＲＮＡ拷贝数（ＡＢＬ做内参），较初诊时降低＞３个ｌｏｇ者预后良好。究竟在诱导后还是巩固后降低＞３个ｌｏｇ较有预后意义，视不同方案而定。如果诱导结束后降低不足３个ｌｏｇ，在巩固期间继续降低，直到治疗完全结束后＜初诊时的４个ｌｏｇ者预后仍好，提示连续监测的重要性。但如果将ＲＵＮＸ１ＲＵＮＸ１Ｔ１和ＣＢＦＢＭＹＨ１１分开统计，可能有不同的结果，因此需后续更多的研究［１０］。关于ＫＭＴ２Ａ（ＭＬＬ）基因重排，在ＡＭＬ只有ＭＬＬＴ３ＫＭＴ２Ａ／ｔ（９；１１）有详细研究［４］，结果显示诱导缓解后ＲＴｑＰＣＲ＜１０－３和后续监测仍＜１０－３复发率低。ＢＣＲＡＢＬ融合基因在ＡＭＬ发生率低，用于ＭＲＤ监测的意义尚未有足够的临床验证。［１１］以上研究结果提示在用融合基因检测ＭＲＤ方面，连续监测的重要性，治疗结束后仍低度阳性不影响预后。２２突变基因不是所有与ＡＭＬ相关的突变基因都适用于ＭＲＤ监测。［１，４５，１０１１］造血干细胞基因突变后，获得竞争优势导致克隆造血，但仍有多系分化和成熟的造血功能不属于白血病细胞，只有在继发其他突变后才出现分化阻滞和无限增殖，发展为白血病。随年龄增长干细胞的突变率增加，因此克隆造血主要见于老年人，儿童少见。较常见的克隆造血突变基因是编码表观遗传修饰因子的基因ＤＮＭＴ３Ａ、ＴＥＴ２、ＩＤＨ、ＡＳＸＬ１，以及ＲＮＡ剪接子和Ｃｏｈｅｓｉｎｓ蛋白的基因等。胚系突变基因ＲＵＮＸ１、ＧＡＴＡ２、ＣＥＢＰＡ、ＤＤＸ４１和ＡＮＫＲＤ２６也不适用于ＭＲＤ检测。与治疗前比较，缓解后突变基因的表达水平仍没有明显下降，需考虑是克隆造血突变或胚系突变基因。胚系突变基因可通过检测胚系组织／细胞（例如口腔黏膜等）ＤＮＡ发现。

海洋微生物活性物质(精)

海洋微生物活性物质的研究进展专业:生物工程姓名:李振森学号:4012010302海洋是生命的发源地,约占地球表面积的71%,其中生物种类20多万种,其多样性远远超过陆地生物的多样性。

由于海洋环境具有高盐度、高压、低营养、低温和无光照等条件,从而形成了海洋生物与陆地生物不同的生长方式和代谢系统。

近年来,随着人们对海洋生物研究的不断深入,发现了多种多样的生物及许多具有新颖、特异化学结构的生物活性物质。

海洋生物活性物质主要包括生物信息物质、生理活性物质、海洋生物毒素及生物功能材料等。

目前,从海洋生物中已相继发现300余种新型化合物,结构新颖并具有多样性:有枯类、聚醚类、当醇类、皂昔类、生物碱、多糖、小分子肤、核酸及蛋白质等,并具有丰富的生理及药理活性,包括抗菌、抗肿瘤、抗病毒、防治心血管疾病、延缓衰老及免疫调节等多种功能。

多年来,国内外一直致力于这方面的研究,试图从中开发结构明确,疗效肯定的新型生物活性物质,以用于攻克人类面临的重大疑难疾病,其中具有高生物活性和高选择性的海洋生物毒素备受重视,成为研究的热点。

近年来,海洋生物毒素是海洋生物活性物。

1、海洋抗肿瘤活性物质1.1海洋放线菌海洋有着极其丰富的放线菌资源,具有抗菌活性的海洋微生物中约有45%来源于放线菌。

就目前的报道,海洋放线菌产生的活性物质大部分来源于小单孢菌属和链霉菌属。

由于海洋放线菌所产生的代谢产物具有功能独特、结构新颖等特点而受到人们的广泛关注,例如抗真菌、抗疟等功能。

另一方面,陆生放线菌的不断开发,发现新的活性物质的可能性越发减少,迫使人们将目光转向海洋放线菌的开发。

1991年Fenical小组[1]首次发现一属全新的需盐生长的特殊海洋放线菌Salinispora,其广泛存在于热带和亚热带海泥中。

2003~2005 年Fenical小组从菌株Salinispora tropica CNB-392 中分离得到10个结构新颖的化合物[2-4],其中化合物Salinosporamide A(1[3]具有广阔的成药前景,对人结肠癌细胞的IC50为0.035 nmol/L,已作为癌症药物进入临床前研究[5-6]。

Identification by HPLC-MS of Carotenoids of the Thraustochytrium CHN-1 Strain Isolated from the Seto

Biosci.Biotechnol.Biochem.,67(4),884–888,2003NoteIdentiˆcation by HPLC-MS of Carotenoids of the Thraustochytrium CHN-1Strain Isolated from the Seto Inland SeaMarvelisa L.C ARMONA ,1Takeshi N AGANUMA ,1and Yukiho Y AMAOKA 2,†1Schoolof Biosphere Sciences,Hiroshima University,1–4-4Kagamiyama,Higashi-Hiroshima 739–8528,Japan2National Institute of Advanced Industrial Science and Technology,2–2-2Hirosuehiro,Kure,Hiroshima 737–0197,JapanReceived August 12,2002;Accepted December 5,2002The orange-pigmented Thraustochytrium ,CHN-1strain was found to contain astaxanthin as the main carotenoid pigment.Echinenone,canthaxanthin,phoenicoxanthin and b -carotene were also identiˆed by high-performance liquid chromatography (HPLC)and HPLC-mass spectrometry.The total extractable carote-noid level was found to increase with culture age.Key words:Thraustochytrium ;protist;carotenoids;astaxanthinThraustochytrids are a group of aquatic hetero-trophic protists commonly found in the marine and estuarine environment.1–3)Characterized by the production of heterokont zoospores and a cell wall consisting of thin golgi-derived non-cellulosic scales,this group has recently been taxonomically classiˆed with the stramenopile clades of protists under the phylum Labyrinthulomycota.4–7)However in the popular or colloquial sense,thraustochytrids are con-sidered to be true fungi.8)The capacity of thraustochytrids to produce a large amount of polyunsaturated fatty acids (PUFA)such as docohexanoic (C22:6DHA)and eicosapentanoic (C20:5,EPA)acids has drawn scientiˆc and com-mercial interest.9)PUFA have been recognized as an important dietary supplement in preventing blood platelet aggregation and decreasing the blood cholesterol level,thereby reducing the risk of arteriosclerosis.10)PUFA have also been found to be eŠective in retarding the growth of tumor cells.11)The presence of carotenoid pigments among the thraustochytrid group of protists has been reported such as the case of the species,Schizochytrium aggregatum ,in which the ketocarotenoids,echine-none and cantaxanthin were identiˆed.12)Carotenoids,a major class of natural pigments,are known to play crucial roles in human nutrition.Their antioxidative property is believed to be respon-sible for protecting against free-radical mediated damage from degenerative disorders such as epitheli-al cancer,cardiovascular disease and age-related macular degeneration thus the intake of a carotene-rich diet is associated with a reduced risk of can-cer.13,14)These beneˆcial biological activities have prompted an increased demand for carotenoid com-pound from sources that have a renewable character-istic and natural high yielding property for the production of a safe colorant for food and phar-maceuticals.As a food colorants,carotenoid are incorporated as the feed supplement for poultry and aquaculture products to impart a desirable color of the skin,fat and other tissues of these farmed animals.15)Taking into account the biosynthetic potential of this microscopic group of protists in producing two equally important compounds,that is,PUFA and carotenoids,the chromatic thraustochytrid strains capable of producing these two compounds were screened.We report in this communication the iden-tiˆcation of carotenoids from an orange-colored thraustochytrid strain by using high-performance liquid chromatography-mass spectrometry (HPLC-MS).Thraustochytrid CHN-1,a docohexanoic acid (C22:6DHA)and carotenoid-producing strain,was isolated from a seawater sample collected from the coastal area of Nagahama (Hiroshima prefecture)in the Seto Inland Sea of Japan.The pine pollen baiting methods of Gaertner 16)was used with modiˆplete identiˆcation of the organism will be reported elsewhere.Pure slant cultures of the Thraustochytrid CHN-1,were grown for 48hours on a GYP medium contain-ing 20g of glucose,10g of peptone,5g of yeast ex-tract,and 20g of agar per liter of a half salt concen-tration of seawater.A loopful each of the vegetative cells was inoculated into a 100-ml Erlenmeyer ‰asks each containing 30ml of a broth medium made up of 2z (w W v)glucose,0.3z (w W v)yeast extract,0.07z (w W v)corn steep liquor (Wako Pure Chemical Indus-tries,Osaka,Japan)and 0.3z (w W v)KH 2PO 4.TheFig.1.Transmission Electron Micrograph of Thraustochytrium sp.CHN-1.Zoosporoblast Containing four Well-developed Zoospores Each within Its Own Membrane Wall;Dark Lipid Bodies Can Be Seen within the Zoospores.885Identiˆcation by HPLC-MS of Carotenoids of the Thraustochytrium CHN-1Strain Isolated from the Seto Inland Sea liquid culture was grown under low intensity ‰uores-cent light (1500lux)with rotary shaking at 120rpm and 239C.At various times,from 3days to the 14days of culture,the carotenoid content of each of the three randomly selected ‰asks was determined.The cells in the liquid cultures were harvested by centrifu-gation at 2,000×g for 20min,and the supernatant was then discarded.The cell pellets were rinsed twice with 100ml of cold reconstituted seawater freeze-dried with a Yamato Neocool freeze-dryer,and then stored at －209C prior to extraction.The lyophilized cells were moistened with water to give a thick paste and extracted quickly with chloroform-methanol (3:1).The extraction procedure was repeated at least three times until the cell debris was almost colorless.All extraction and subsequent manipulation of the samples were conducted in subdued light.The sol-vent was removed in vacuo to obtain a crude residue of the extract.This residue was dissolved in 1ml of chloroform,and the solution was applied to a column (F 1.0cm ×F 10cm)of silica gel that had been washed and packed by using hexane.After eluting with chlo-roform,the fraction was collected and dried in vacuo .The residue was reconstituted with methanol,and the resulting solution was loaded separately into two diŠerent HPLC instruments each equipped with a photodiode array detector (Shimadzu SPD-10MAVvp).The ˆrst HPLC instrument was capable of detect-ing UV-visible wavelength carotenoid spectra,from which the retention time and quantiˆcation of each individual carotenoid was calculated.The second HPLC-MS system allowed the detection of the sig-niˆcant masses of the carotenoids as well as the UV-visible light absorption spectra (UV-VIS),from which the ˆnal identity of the carotenoids was con-ˆrmed.HPLC-MS was conducted with a Shimazu LCMS-2010instrument.The carotenoid solution was ˆltered through Ekicrodisc R 3CR ˆlters (0.45m m ×3mm)and separated in a Shim-pack VP-ODS column (2.0mm I.D.×150mm)at 259C.The mobile phase consisted of solvent A (water)and solvent B (methanol),the following gradient procedure being used:50z of B for 0min;a linear gradient from 0to 100z of B for 15min;100z of B for 35min.The‰ow rate was 0.2ml Wmin,and samples were ionized by APCl (＋).To obtain pure carotenoid isolates for quantiˆca-tion purposes,the crude residue extract was dissolved in a small volume of acetone,and the resulting solu-tion was applied to a column (F 1.0cm ×F 10cm)of silica gel that has been washed and packed by using hexane.Carotene-like b -carotene was eluted with hexane,and the xanthophylls such as echinenone and canthaxanthin were recovered by eluting with 75z (3:1)hexane in chloroform,while astaxanthin andphoenicoxanthin were eluted with equal volumes of hexane and chloroform (1:1).Each solution contain-ing the carotenoid fractions from the silica gel column was dried in vacuo ,and further puriˆed by high-performance liquid chromatography (HPLC)in a Wakosil 5C18analytical column (q 4.6mm ×150mm)until the chromatographic purity of each individual carotenoid had been achieved.HPLC was conducted with a Shimadzu LC-10AS instrument with the detector set at 450nm.The carotenoids were eluted isocratically with a solvent mixture of methanol (90z ),water (8z ),and acetonitrile (2z )at a ‰ow rate was set at 3.0ml W min.Quantitative data for each individual carotenoid were based on a comparison of the peak area with known standards (astaxanthin,canthaxanthin,echinenone,and b -carotene).In the absence of a known standard such as in the case of the carotenoid,phoenicoxanthin,the astaxanthin peak area was used to calculate its concentration.Figure 1shows a transmission electron micrograph of Thraustochytrid CHN-1strain vegetative cells.Thraustochytrium strains are known to form a scale-like cell wall,and the vegetative cells in this species formed scales covering the plasma membrane.Our sequence analysis data for DNA (18S rDNA;data not shown)provided additional evidence to substan-tiate our claim that the isolated organism was indeed a Thraustochytrium.Many of the studies on carotenoid analysis here886Fig.2.HPLC Proˆle of the Carotenoids from Thraustochytrium CHN-1after14Days of Cultivation.Astaxanthin1,phoenicoxanthin2,canthaxanthin3,echinenone4,and b-carotene5.A Shim-pack VP-ODS column(2.0mm I.D.150mm)was used at259C.The mobile phase consisted of solvent A(water)and solvent B(methanol).The following gradient procedure was used:50z of B for0min;a linear gradient from0to100z of B for15min;100z of B for35min.The‰ow rate was0.2ml W min.and a photodiode array detector was used.See Table1for identiˆcation of the peaks.Table1.Identiˆcation of Carotenoids in Traustochytrium sp.CHH-1after14Days of Cultivation by LCPeak no.RetentiontimeAbsorptionmaximum(nm)Mass(m W z M＋)(Analyzed by APCI＋)Compound1 3.2486597Astaxanthin2 4.5482581Phoenicoxanthin3 5.8481565Canthaxanthin 413.3470551Echinenone 535.2465,493537b-CaroteneM.L.C ARMONA et al.attempted to identify(under isocratic conditions)the major carotenoids and minor pigments(for which standards may be available),based on the HPLC retention times characteristics,UV-visible maxima and probably the nature of the column packing or the stationary phase used.However establishing the carotenoid identity based on standard spectra from gradient elution is rather di‹cult.Figure2show typical chromatographic data for each individual carotenoid obtained from the Thraustochytrid CHN-1strain.This chromatographic data was obtained from a photodiode array monitor,and we plotted the molecular mass of each recorded peak andˆnally resolved the identity of each individual carotenoid.The quantitative composition of the in-dividual carotenoids extracted from the cell pellets of Thraustochytrium CHN-1presented in Table1are as follows:astaxanthin,(peak no.1),echinenone(4), canthaxanthin(3),phoenicoxanthin(2),and b-caro-tene(5)in order of the retention time.In the case of two carotenoids having the same molecular mass i.e.,a-carotene,b-carotene and g-carotene,the column chromatographic patterns and HPLC retention times were matched with the mass spectral data to reinforce the analysis.The major carotenoid peak from Thraustochytrium CHN-1was identiˆed as astaxanthin,and was further analyzed by checking its CD spectrum.The CD spectrum (Fig.3),suggesting a3S end group of astaxanthin,is consistent with that of the optically pure(3S,3?S-astaxanthin)reported by Englert,(1977).17) The growth and carotenoid accumulation are shown in Fig. 5.The carotenoid content of Thraustochytrid CHN-1paralleled the biomass or cell growth,the maximum stationary phase of growth being reached after8days of incubation.Fur-ther incubation to14days resulted in no further in-crease in either the cell biomass or the total extracta-ble carotenoids,a fall instead being apparent.The total content of b-carotene in Thrausto-chytrium CHN-1was found to have declined by the8th day of culture while the astaxanthin content increased.Our observation agrees with the general observation of others,18–20)that the total amount of carotenes(hydrocarbon carotenoids)was substantial-ly lower during the late or stationary phase,while the xanthophylls content(oxygenated carotenoids)was higher.The observation of the sequential changes in the total content of b carotene and that of the ketocarotenoids,echinenone,phoenicoxanthin and canthaxanthin strongly suggests for the possible inter-conversion of these carotenoids in Thrausto-chytrium CHN-1.Donkin,(1976),21)working on the ketocarotenoid biosynthesis in Haematococcus lacus-Fig.3.CD Spectrum in Dichloromethane 2.12×10－5M of 3S,3?S trans -Astaxanthin from ThraustochytriumCHN-1.Fig.4.Chemical Structuresof the Carotenoid Isolatedfrom Thraustochytrium CHN-1.Fig.5.Growth Curve and Carotenoid Accumulation from Thraustochytrium CHN-1Grown for 14Days.line indicates growth curve,indicates astaxanthin,indicates phoenicoxanthin,indicates canthaxanthin,indicates b -carotene.887Identiˆcation by HPLC-MS of Carotenoids of the Thraustochytrium CHN-1Strain Isolated from the Seto Inland Sea tris ,has demonstrated the same characteristics and concluded that the sequential changes in the concen-tration of the ketocarotenoids suggests their possible role as intermediates on the pathway to astaxanthin,thus,the conversion of b -carotene to astaxanthin occurs via these ketocarotenoids like canthaxanthin and echinenone.To check the sequence of intermedi-ates on the pathway to astaxanthin in Thrausto-chytrium CHN-1,we used diphenylamine,a com-pound that inhibits the insertion of a keto group into the carotenoid molecule.Our preliminary results demonstrate that (3S,3?S)astaxanthin was synthe-sized in Thraustochytrid CHN-1from b -carotene via echinenone and canthaxanthin (to be reported else-where).However,our results do not seem to cor-roborate the results obtained by Fan (1999)22)for the green alga Haematococcus pluvialis (data not shown).The eŠect of variation in a number of parameters like the physical factors (pH and temperature)and nutritional factors (carbon and nitrogen sources)during the culture can alter the relative quantities of carotenoids produced.19,23)However in this present work no attempt was made to assess the qualitative variation in the carotenoid content of Thrausto-chytrium CHN-1under varying environmental con-ditions.The results presented here represent the ˆrst evidence of the Thraustochytrium CHN-1strain pos-sessing the capacity to produce the highly oxidized888M.L.C ARMONA et al.form of a carotenoid,astaxanthin.Among the organisms reportedly found to synthesize a high amount of astaxanthin are the green algae Hae-matococcus pluvialis,24)yeast Pha‹a rhodozyma,18) and Agrobacterium aurauticum.20)Our current screening of chromatic Thrausto-chytrium strains involves two important criteria:one is its ability to produce DHA and the other is its abil-ity to synthesize carotenoids,speciˆcally astaxanthin. Su‹ce it to say,therefore that the appreciable amount of astaxanthin in Thraustochytrium CHN-1 warrants its applicability for pilot-plant-scale production and that if astaxanthin production is cou-pled with PUFA production,it will be beneˆcial to industry.Our future endeavor is geared towards optimizing the production of the two compounds, astaxanthin and PUFA,in this isolate. References1)Raghu-kumar,S.,and Gaertner,A.,Ecology of thethraustochytrids(lower marine fungi)in the Fladen Ground and other parts of the North Sea II.VeroŠ.Inst.Meeresforsch.Bremerh.,18,298–308(1980). 2)Naganuma,T.,Takasugi,H.,and Kimura,H.,Abundance of thraustochytrids in coastal plankton.Mar.Ecol.Prog.Ser.,162,105–110(1998).3)Kimura,H.,Fukuba,T.,and Naganuma,T.,Bio-mass of thraustochytrid protoctists in coastal water.Mar.Ecol.Prog.Ser.,189,27–33(1999).4)Patterson, D.J.,Stramenophiles:chromophytesfrom a protistant perspective.Systematics Associa-tion,Special Volume38,Clarendon Press,Oxford, 357–379(1989).5)Cavalier-Smith,T.,Allsopp,M.T.E.P.,and Chao,E.E.,Thraustochytrids are chromists not fungi:sig-nature sequences of heterokonta.Phil.Trans.Roy.Soc.Lond.Biol.Science,346,387–397(1994).6)Leipe,D.D.,Tong,S.M.,Goggin,C.L.,Slemenda,S.,Oieniazek,N.J.,and Soggin,M.L.,16S-like rDNA sequences from Developayella elegans, Labyrinthuloides haliotis,and Protermonas lacerate conˆrm that the stramenophiles are primarily a heterotrophic group.Eur.J.Protistol.,32,449–458 (1996).7)Darley,W.M.,Porter,D.,and Fuller,M.S.,Cellwall composition and synthesis via Golgi-directed scale formation in the marine eukaryote Schizo-chytriun aggregatum,with a note on Thraustochytri-um sp.Arch.Mikrobiol.,90,89–106(1973).8)Barr,D.J.S.,Evolution and kingdoms of organismsfrom the perspective of a mycologist.Mycologia,84, 1–11(1992).9)Lewis,T.E.,Nichols,P.D.,and McMeekin,T.A.,The biotechnological potential of thraustochytrids.Mar.Biotechnology,1,580–587(1999).10)Bang,H.O.,and Dyerberg,J.,Plasma andlipoproteins in Greenlandic West Coast Eskimos.Acta Med.Scand.,192,85–94(1972).11)Bang,H.O.,and Dyerberg,J.,Lipid metabolism andischemic heart disease in Greenland Eskimos.Adv.Nutr.Res.,3,1–21(1986).12)Valadon,L.R.G.,Carotenoids as additionaltaxonomic characters in fungi:a review.Trans.Br.Mycol.Soc.,67,1–15(1976).13)Seddon,J.M.,Ajani,U. A.,Sperduto,R. D.,Hiller,R.,Blair,N.,Burton,T.C.,Farber,M.D., Gragoudas, E.S.,Haller,J.,and Miller, D.T., Dietary carotenoids,vitamins A,C and E and advanced age related macular degeneration.JAMA, 272,1413–1420(1994).14)Willet,W.C.,and Trichopoulos,D.,Nutrition andcancer:A summary of the evidence.Cancer Causes Control,7,178–180(1996).15)Johnson,E.A.,and An,G.H.,Astaxanthin frommicrobial sources.Critical Rev.Biotechnology,11, 297–326(1999).16)Gaertner,A.,Eine methode des nachweises niederermit pollen koderbarer pilze im meerwasser und im sediment.VeroŠ.Inst.Meeresforch.Bremer.Sonderb.,3,75–92(1968).17)Englert,G.,Kienzle,F.,and Noack,K.,1H-NMR-,13C-NMR-,UV.-und CD-daten von synthetischen (3S,3?S)-astaxanthin,seinen15-cis Isomeren und einigen analogen verbindungen.Helv.Chim.Acta, 60,1209–1212(1977).18)Andrews, A.G.,PhaŠ,H.J.,and Starr,M.P.,Carotenoids of Pha‹a rhodozyma,a red pigmented fermenting yeast.Phytochemistry,15,1003–1007 (1976).19)Johnson, E. 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CurriculumVita-Chiao-Yao（Joe）She-CAS

Curriculum Vita - Chiao-Yao (Joe) SheEducation:1957 - B.S., Taiwan University, Taipei, Taiwan1961 - M.S., North Dakota State University, Fargo, North Dakota1964 - Ph.D., Stanford University, Stanford, CaliforniaExperience:1975-Present Professor of Physics, Colorado State University, Fort Collins, CO1968- 1971 Assistant Professor of Physics, Colorado State University1971- 1975 Associate Professor of Physics, Colorado State University1964 - 1968 Assistant Professor of Electrical Engineering, The University of Minnesota Honors, Memberships and Services:Fellow of the Optical Society of AmericaMember of APS, AGU1976 Research Publication Award, Naval Research Laboratory, Washington, D.C.1978 President of the Rocky Mountain Section of OSA.1987 Burlington Northern Faculty Achievement Award, Colorado State University 1988-1989 Golden Screw (Teaching) Award, Colorado State University1988, 1995 On NSF Review Panels for Research Initiation Awards, Lightwave Technology Program, and Optical Science and Engineering, respectively1992-1994 Member, AMS Committee on Laser Studies of the Atmosphere1993-1995 Member, CLEO/IQEC Program Committee1994-1997 Member, Arecibo Users and Scientific Advisory Committee, N.A.I.C.1989-2005 Fellow, Coop. Institute for Research in the Atmosphere, Colorado State University1997-2000 Member, NSF CEDAR Science Steering Committee2000-2001 Fulbright Research Award, Norway2003NSF/CEDAR Workshop – CEDAR Lecture Prize2003AGU Editor’s Citation – Outstanding Reviewer for Geophysics Research Letters 2005 Included in the 60th Diamond Edition of Marquis Who’s Who in AmericaBook Chapter and EditingWilliam B. Grant, Edward V. Browell, Robert T. Menzies, Kenneth Sassen and Chiao-Yao She (Editors), Selected Papers on Laser Applications in Remote Sensing, SPIE Milestone Series, MS 141 (1997).Research Interest and Current SupportThe research interests of Prof. She have been broad and often interdisciplinary. He has developed new laser measurement techniques for solving basic as well as applied problems. For the past 25 years, he has made a series of innovations in two high-spectral-resolution lidars: Rayleigh-Mie lidar and narrowband sodium lidar for atmospheric temperature and wind measurements, respectively in the lower and upper atmosphere. The narrowband sodium lidar, now capable of measuring mesopause region (80-110km in altitude) temperature and wind on 24-hour continuous basis, has enjoyed considerable success with continued NSF funding for upper atmospheric research since 1989. Its technology and innovations were and are beingduplicated by National and International Middle and Upper Atmospheric Research Facilities. In the past ten years, Prof. She’s research has been supported by NSF, NASA and AFOSR.Research PublicationsProfessor She has co-authored about 170 papers in refereed journals. A selected list since 1998:109. She, C. Y. and U. von Zahn, The concept of two-level mesopause: Support through new lidar observation, J.Geophys. Res., 103, 5855 - 5863, 1998.110. She, C. Y., S. W. Thiel and D. A. Krueger, Observed episodic warming at 86 and 100 km between 1990 and 1997: Effects of Mount Pinatubo eruption, Geophys. Res. Lett., 25, 497 - 500, 1998.114. She, C. Y., and R. P. Lowe, Seasonal temperature variations in the mesopause region at mid-latitude: comparison of lidar and hydroxyl rotational temperatures using WINDII/UARD OH height profiles, J. Atmo.Solar-Terr. Physics, 60, 1573-1583, 1998.119. She, C. Y., S. S. Chen, Z. L. Hu, J. Sherman, J. D. Vance, V. Vasoli, M. A. White, J. R. Yu, and D. A.Krueger, Eight-year climatology of nocturnal temperature and sodium density in the mesopause region (80 to 105 km) over Fort Collins, CO (41o N, 105o W), Geophys. Res. Lett., 27, 3289 - 3292, 2000.120. She, Chiao-Yao, Spectral structure of laser light scattering revisited: bandwidths of nonresonant scattering lidar, Appl. Opt. 40, 4875-4884, 2001.124. She, C. Y., Joe D. Vance, B. P. Williams, D. A. Krueger, H. Moosuller, D. Gibson-Wilde, and D. C. Fritts, Lidar studies of atmospheric dynamics near polar mesopause, EOS, Transactions, American Geophysical Union, 83 (27), P.289 and P.293, 2002.125. She, C. Y., Songsheng Chen, B. P. Williams, Zhilin Hu, David A. Krueger and M. E. Hagan, Tides in the mesopause region over Fort Collins, CO (41o N, 105o W) based on lidar temperature observations coveringfull diurnal cycles, Jour. Geophys. Research, 107, N. 0, 10.1029/2001JD001189, 2002.134. She, C. Y., Jim Sherman, Tao Yuan, B. P. Williams, Kam Arnold, T. D.Kawahara, Tao Li, LiFang Xu, J. D.Vance and David A. Krueger, The first 80-hour continuous lidar campaign for simultaneous observation of mesopause region temperature and wind, Geophys. Res. Lett.30, 6, 52, 10.1029/2002GL016412, 2003.136. She, C. Y., and D. A. Krueger, Impact of natural variability in the 11-year mesopause region temp erature observation over Fort Collins, CO (41N, 105W), Adv. Space Phys. 34, 330-336, 2004.137. Beig, G.; Keckhut, P.; Lowe, R. P.; Roble, R. G.; Mlynczak, M. G.; Scheer, J.; Fomichev, V. I.; Offermann,D.; French, W. J. R.; Shepherd, M. G.; Semenov, A. I.; Remsberg,E. E.; She, C. Y.; Lübken,F. J.; Bremer, J.;Clemesha, B. R.; Stegman, J.; Sigernes, F.; Fadnavis, S. (2003), Review of mesospheric temperature trends,Rev. Geophys., Vol. 41, No. 4, 1015, 10.1029/2002RG000121.138.Chiao-Yao She, Initial full-diurnal-cycle mesopause region lidar observations: Diurnal-means and tidal perturbations of temperature and winds over Fort Collins, CO (41N, 105W), PSMOS 2002, J. Atmo. Solar-Terr. Phys. 66, 663-674, 2004.139. She, C. Y.,Tao Li, Biff P. Williams, Tao Yuan and R. H. Picard (2004), Concurrent OH imager and sodium temperature/wind lidar observation of a mesopause region undular bore event over Fort Collins/Platteville, CO, J. Geophys. Res. 109, D22107, doi:10.1029/2004JD004742.140. She, C.Y., T. Li, R. C. Collins, T. Yuan, B. P. Williams, T. D. Kawahara, J. D. Vance, P. Acott, D. A. Krueger,H.-L. Liu, and M. E. Hagan (2004), Tidal perturbations and variability in the mesopause region over FortCollins, CO (41N, 105W): Continuous multi-day temperature and wind lidar observations, Geophys. Res. Lett., 31, L24111, doi:10.1029/2004GL021165.142. Fritts, D. C., B. P. Williams, C. Y. She, J. D. Vance, r. Rapp, F.-J. L¨ubken, A. F. J. Schmidlin, A. M¨ullemann R. A. Goldberg (2004) Observations of extreme temperature and wind gradients near the summer mesopause during the MaCWAVE/MIDAS rocket campaign, Geophys. Res. Lett., 31, L24S06, doi:10.1029/2003GL019389.144. Tao Li, C. Y. She, Bifford P. Williams, Tao Yuan, Richard L. Collins, Lois M. Kieffaber and Alan W.Peterson (2005), Concurrent OH imager and sodium temperature/wind lidar observation of localized ripples over Northern Colorado, J. Geophys. Res. 110, D13110, doi:10.1029/2004JD004885146. Chiao-Yao She (2005), On atmospheric lidar performance comparison: from power aperture to power–aperture–mixing ratio–scattering cross-section, Modern Optics, 52, 2723-2729, DOI:10.1080/09500340500352618.147. She, C. Y., B. P. Williams, P. Hoffmann, R. Latteck, G. Baumgarten, J. D. Vance, J. Fiedler, P. Acott, D. C.Fritts, F.-J. Luebken (2006): Observation of anti-correlation between sodium atoms and PMSE/NLC in summer mesopause at ALOMAR, Norway (69N, 12E), J. Atmos. Solar-Terres. Phys. 68, 93-101.148. Yuan, T., C. Y. She, M. E. Hagan, B. P. Williams, T. Li, K. Arnold, T. D. Kawahara, P. E. Acott, J. D. Vance,D. A. Krueger and R. G. Roble (2006), Seasonal variation of diurnal perturbations in mesopause-regiontemperature, zonal, and meridional winds above Fort Collins, CO (40.6°N, 105°W), J. Geophys. Res. 111, D06103, doi:10.1029/2004JD005486.149. Xu, Jiyao, C. Y. She, Wei Yuan, C. J. Mertens, Marty Mlynzack, J. R. Russell (2006), Comparison between the temperature measurements by TIMED/SABER and Lidar in the midlatitude, J. Geophys. Res., 111, A10S09, doi:10.1029/2005JA011439.150. Xu, J., A. K. Smith, R. L. Collins, and C.-Y. She (2006), Signature of an overturning gravity wave in the mesospheric sodium layer: Comparison of a nonlinear photochemical-dynamical model and lidar observations, J. Geophys. Res., 111, D17301, doi:10.1029/2005JD006749.151. Sherman, J. P., and C.-Y. She (2006), Seasonal variation of mesopause region wind shears, convective and dynamic instabilities above Fort Collins, CO: A statistical study, J. Atmo. Solar-Terr. Physics, 68, 1061-1074. 154. Davis, D. S., P. Hickson, G. Harriot and C. Y. She (2006), Temporal variability of the telluric sodium layer, Optics Lett. 31, 3369-3371.155. She, Chiao-Yao, J. D. Vance, T. D. Kawahara, B. P. Williams, and Q. Wu (2007), A proposed all-solid-state transportable narrowband sodium lidar for mesopause region temperature and horizontal wind measurements, Canadian Journal of Physics, 85, 111 – 118.156. Gumbeli, J., Z. Y. Fan, T. Waldemarsson, J. Stegman, G. Witts, E. J. Llewellyn, C.-Y. She and J. M. C. Plane (2007), Retrieval of global mesospheric sodium densities from the Odin satellite, Geophys. Res. Lett.,. 34, L04813, doi:10.1029/2006GL028687.157. Li, T., C.-Y. She, H.-L. Liu, and M. T. Montgomery (2007), Evidence of a gravity wave breaking event and the estimation of the wave characteristics from sodium lidar observation over Fort Collins, CO (41_N, 105_W) , Geophys. Res. Lett.,. 34, L05815, doi:10.1029/2006GL028988.158. She, C.-Y., J. Yue and Z.-A. Yan, J. W. Hair, J.-J. Guo, S.-H. Wu and Z.-S. Liu (2007), Direct-detection Doppler wind measurements with a Cabannes-Mie lidar: A. Comparison between iodine vapor filter and Fabry-Perot interferometer methods, Applied Optics 46, 4434-4443.159. She, C.-Y., J. Yue and Z.-A. Yan, J. W. Hair, J.-J. Guo, S.-H. Wu and Z.-S. Liu (2007), Direct-detection Doppler wind measurements with a Cabannes-Mie lidar:B. Impact of aerosol variation on iodine vapor filter methods, Applied Optics 46, 4444-4454.161. Tao Li, C.-Y. She, Scott E. Palo, Qian Wu, Han-Li Liu, and Murry L. Salby (2008), Coordinated Lidar and TIMED observations of the quasi-two-day wave during August 2002-2004 and possible quasi-biennial oscillation influence, Advanced Space Research 41, 1462-1470.162. Liu, H.-L., T. Li, C.-Y. She, J. Oberheide, Q. Wu, M. E. Hagan, J. Xu, R. G. Roble, M. G. Mlynczak, and J. M.Russell III (2007), Comparative study of short term diurnal tidal variability, J. Geophys. Res.(In press).163. She, Chiao-Yao, and David A. Krueger (2007), Laser-Induced Fluorescence: Spectroscopy in the Sky, Optics & Photonic News (OPN), 18(9), 35-41.164. Li, T., C. -Y. She, H.-L. Liu, T. Leblanc, and I. S. McDermid, Sodium lidar observed strong inertia-gravity wave activities in the mesopause region over Fort Collins, CO (41°N, 105°W) J. Geophys.Res. 112, D22104, doi:10.1029/2007JD008681.165. Yuan T., C.-Y. She and D. A. Krueger, F. Sassi, R. Garcia, R. Roble, H.-L. Liu, and H. Schmidt (2008), Climatology of mesopause region temperature, zonal wind and meridional wind over Fort Collins, CO (41ºN, 105ºW) and comparison with model simulations, J. Geophys. Res. 113, D03105, doi:10.1029/2007JD008697. 166. Liguo Su, R. J. Coolins, D. A. Krueger and C.-Y. She, Statistical Analysis of Sodium Doppler Wind-Temperature Lidar Measurements of Vertical Heat Flux, J. Atm. Oceanic Tech. (In press).167. Yuan, T., C. Y. She, Hauke Schmidt, David A. Krueger, Steven Reising, Seasonal variations of semidiurnal tidal-period perturbations in mesopause region temperature, zonal and meridional winds above Fort Collins, CO(40.6°N, 105°W), J. Geophys. (In oress).168. Yue, J., S. L. Vadas2, C-Y She, et al. (2008), A study of OH imager observed concentric gravity waves near Fort Collins on May 11, 2004, Geophys. Res. Lett. (submitted).169. Vadas, S. L., J. Yue, C.-Y. She and P. Stamus (2008), The effects of winds on concentric rings of gravity waves from a thunderstorm near Fort Collins in May 2004, J. Geophys.Res., (submitted).。

leptospira interrogans ss. icterohaemorrhagiae-感染的

LEPTOSPIRA INTERROGANS SSP.Aetiology Epidemiology Diagnosis Prevention and ControlPotential Impacts of Disease Agent Beyond Clinical Illness References AETIOLOGYClassification of the causative agentLeptospira interrogans, the causative agent of leptospirosis, consists of numerable serotypes capable of causing a variety of disease manifestations in a wide range of hosts. This motile and flagellated spirochete bacterium is recognizable via microscopy and earned its name from a characteristic hooked appearance that resembles a question mark. The taxonomic classifications within the Leptospira genus have been reorganized many times in accordance with new antigenic, genomic, and pathologic data. What was once over 250 serovars grouped into two Leptospira species is now 21 genomospecies of Leptospira with reclassified serovars. While the new taxonomic names are typically used in the scientific literature, historical names still circulate on product labels and in common use.Not all serovars of Leptospira are pathogenic, and many are associated with a reservoir species in which little disease is apparent. Many serovars are highly prevalent within maintenance host populations and persist in the kidneys or genital tract. Small antibody responses and low tissue burdens are typical in these animals. Incidental hosts, however, typically develop serious disease with high tissue burdens and robust antibody responses. These classifications are not always entirely distinct and some overlap between presentations can exist.Leptospirosis is a zoonotic disease but is typically associated with at-risk occupations (veterinarian, livestock owners, dairy workers, etc.) and exposure to contaminated water.Resistance to physical and chemical actionTemperature: Generally fails to persist at <10°C or >34°C;pasteurization and moist heat at 121°C/15 minutes are effective methodsof killing leptospirespH: Prefers neutral to slightly alkaline conditionsChemicals/Disinfectants: Inactivated by 1% sodium hypochlorite, 70%ethanol, formaldehyde, detergents, quaternary ammonium compounds,iodine based compounds, glutaraldehyde, and hydrogen peroxideSurvival: Warm, moist conditions greatly enhance survival; maypersist up to 6 weeks under favorable conditions; freezing, dehydration,and UV radiation inactivate leptospiresEPIDEMIOLOGYHosts●Virtually all mammals are vulnerable to pathogenic Leptospira serovars to varying degrees○Many serovars have specific maintenance hosts while others are more promiscuous●Leptospira serovars known to cause disease in mammals have been isolated from amphibians○Some serovars have also been isolated from invertebrates, reptiles, and birdsProminent host-serovar associations●Armadillos (Dasypus novemcinctus, Euphractus sexcinctus) - Autumnalis, Cynopteri, Hebdomadis,Pomona●Bandicoots (Isoodon macrourus, Perameles spp.) - numerous serovars have been associated withthese species●Brazilian tapir (Tapirus terrestris) - Pomona●Canids (Canis latrans, C. familiaris, C. lupus) - Bratislava, Canicola, Grippotyphosa, Hardjo,Icterohaemorrhagiae, Pomona●Cattle (Bos taurus, Syncerus caffer) - Hardjo and others●Cervids - Bratislava, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, Pomona●European hedgehog (Erinaceus europaeus)●Felids (Felis silvestris silvestris, F. silvestris catus, Lynx spp.)●Flying foxes (Pteropus spp.) - a multitude of Leptospira serovars have been identified in variousspecies of bat●Foxes (Vulpes lagopus, V. vulpes, Urocyon cinereoargenteus, Lycalopex griseus) - Bratislava,Canicola, Grippotyphosa●Giant anteater (Myrmecophaga tridactyla) - Djasiman●Horses (Equus ferus) - Bratislava●Lagomorphs (Lepus europaeus, L. timidus, Oryctolagus cuniculus) - Grippotyphosa●Marine mammals (Eubalaena australis,Trichechus manatus ) - Australis, Manaua○Sea lions (Zalophus californianus, Z. wollebaeki) and seals (Callorhinus ursinus, Phoca vitulina, Mirounga angustirostris, Arctocephalus forsteri) - Canicola, Hardjo, Pomona ○There have been multiple mass-mortality events attributed to leptospirosis in California sea lions●Marsupials - Australis, Autumnalis, Ballum, Bataviae, Celledoni, Cynopteri, Djasiman,Grippotyphosa, Hardjo, Hebdomadis, Icterohaemorrhagiae, Javanica, Mini, Panama, Pomona, Pyrogenes, Sejroe, Tarassov, Topaz●Mongooses (Herpestes auropunctatus, Mungos mungo, Paracynictis selousi) - Bratislava, Hardjo●Mustelids (Meles meles, Martes fiona, M. martes, Mustela putorius, M. nivalis, M. ermine, Lutra lutra)●Platypus (Ornithorhynchus anatinus) - Autumnalis, Hardjo, Grippotyphosa●Raccoons (Procyon itor) and skunks (Mephitis mephitis) - Bratislava, Canicola, Grippotyphosa,Hardjo, Icterohaemorrhagiae, Pomona●Rodents and insectivores - Arborea, Australis, Ballum, Bindjei, Broomi, Canicola, Celledoni,Gryppotyphosa, Icterohaemorrhagiae, Javanica, Mini, Pomona, Pyrogenes, Sejroe, Tarrasovi, Zanoni○Rats are well-appreciated hosts for serotype Icterohaemorrhagiae●Swine (Sus scrofa and other spp.) - Bratislava, Hardjo, Pomona●Vervet monkey (Cercopithecus aethiops sabaeus) - Australis, Grippotyphosa, Javanica●Multiple snake, turtle, toad, and frog species have also been identified as PCR and/or serologicallypositiveTransmission●Ingestion●Contact with mucous membranes or wet, abraded skin●Some serovars can be transmitted venerally or transplacentallySources●Urine●Contaminated soil and water●Placental fluids●Genital secretions●Milk●BloodOccurrenceMany wildlife species are reservoirs for Leptospira and subsequently maintain host-bacterium interactions that do not negatively impact the animal. Over time, selection pressures on the leptospires many change and drive reservoir species relationships to shift accordingly. Therefore, it is important to assess the epidemiology of leptospirosis on a more local level to understand transmission risks and disease impacts.Leptospira is globally enzootic, but disease is more frequently seen in warm and moist environments. This may be seasonal (temperate zones) or more constant (tropical regions). Rainfall encourages persistence of the organism in the environment. Additionally, some serotypes are much more geographically dispersed and others are found in more limited regions.For more recent, detailed information on the occurrence of this disease worldwide, see the OIE World Animal Health Information System - Wild (WAHIS-Wild) Interface [http://www.oie.int/wahis_2/public/wahidwild.php/Index].DIAGNOSISAfter invasion of mucous membranes or damaged skin, there is a 4-20 day incubation period followed by a 7-10 day period of circulation in the bloodstream. Clinical signs of acute leptospirosis depend on the tissues colonized during this period of bacteraemia, the host species, and infecting serovar. A robust antibody response follows and is associated with a declining bacteraemia. Tissues may recover slowly or not at all depending on the degree of damage. Death is possible in severe cases.Incidental hosts maintain the bacterium in renal tubules for days to weeks and shed the organism in urine. Maintenance hosts, however, maintain the bacterium in the renal tubules, genital tract, and/or eyes and shed it in urine or genital secretions for months to years after infection.Clinical diagnosisLeptospirosis is a highly variable, systemic infection and the presentation depends on the infecting serovar, the host species, and the host’s general health and immune status. Maintenance hosts typically do not develop significant clinical disease. Incidental hosts typically experience severe, acute disease secondary to bacterial toxins and inflammatory responses generated by the immune system. Initially, animals may be febrile and anorectic. They may quickly develop signs of haemorrhage and haemolytic anaemia secondary to endothelial damage such as mucosal petechiation, icterus, haemoglobinuria/haematuria, dehydration, vomiting, and colic. Acute renal injury develops rapidly and is a significant contributor to mortality. Pneumonia, meningitis, uveitis, corneal opacification, photosensitization, myalgia, and pancreatitis are also possible.Reproductive disease is often characterised by abortion/stillbirth, mummified fetuses, infertility, blood in milk, or a cessation of milk production. If not aborted, neonates infected transplacentally are typically weak. Maintenance hosts do not develop reproductive disease acutely like incidental hosts, but instead remain subclinical for weeks to months.Lesions●Renal tubular necrosis and suppurative nephritis○Pale, oedematous parenchyma +/- pitting of the serosal surface and capsular adhesions○Subcapsular haemorrhage○Inflammation initially characterised by neutrophils but becomes lymphoplasmacytic○Mixed inflammatory processes are associated with higher mortality rates●Hepatomegaly +/- necrotizing hepatitis○The liver is often friable and discolored in a lobular pattern●Pulmonary haemorrhage●Petechiae and ecchymoses on mucous membranes and internal organs●Horses may develop uveitis with conjunctivitis, corneal oedema, synechia, or cataractsDifferential diagnoses●Ocular disease○Equine recurrent uveitis○Traumatic uveitis/reflex uveitis○Infectious conjunctivitis●Kidney disease○Toxin exposure (e.g., ethylene glycol)○Infectious nephritis, pyelonephritis, glomerulonephritis○Renal tubular acidosis○Nematodes (Stephanurus dentatus, Dioctophyma renale)●Reproductive failure or compromise○Brucellosis○Bovine viral diarrhea virus (BVDV)○Porcine reproductive and respiratory syndrome (PRRS)○Q-Fever (Coxiella burnetii)○Neospora spp.○Tritrichomonas foetus○Mastitis, metritis●Liver disease, icterus, and haemolytic anaemia○Viral hepatitis○Toxin exposure (e.g., heavy metals, anticoagulant rodenticides)○Rickettsial infection○Clostridium haemolyticum, C. perfringens A○Neonatal isoerythrolysis●Bacterial septicaemiaLaboratory diagnosisSamplesFor isolation of agent●Kidney●Blood●Urine●Other grossly affected tissue such as liverSerological tests●Serum●Whole bloodProceduresIdentification of the agent●Silver-stained histopathology slides allow for direct visualization of the organism in renal tubules●Immunohistochemistry (IHC)●Bacterial culture○Because the organism is low-growing, this may take 12-26 weeks○Best available method to determine infecting serovar●Polymerase chain reaction (PCR)○Widely variable protocols○Does not provide serovar-specific resultsSerological tests●Microscopic agglutination test (MAT)○Uses live, regionally common serovars of Leptospira○Requires diagnostic laboratory to maintain live cultures of serovars○Provides quantitative titre level●Antibody capture enzyme-linked immunosorbent assay (ELISA)○Currently used for domestic canines; detects antibodies to LipL32 protein○Results are qualitative (positive/negative) and may yield false positives in the event of prior vaccination●Immunofluorescence assay (IFA)●There is not yet a consensus on what a diagnostic titre for Leptospira should be, therefore pairedacute and convalescent sera are recommended for testing●Caution should be taken when interpreting serology data; antibody titre does not always correspondwith disease stateFor more detailed information regarding laboratory diagnostic methodologies, please refer to Chapter 3.1.12 Leptospirosis in the latest edition of the OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals.PREVENTION AND CONTROLSanitary prophylaxis●Extra precautions should be taken when cleaning areas frequented by potential Leptospira hosts.Wear gowns, shoe covers, and gloves to prevent contamination of personal clothing. Face shields are recommended to protect mucous membranes from aerosols.Medical prophylaxis●There are a variety of Leptospira vaccines available for domestic animals, including livestock○Vaccine intent may vary from prevention of infection to reduction of renal colonization and urine shedding○Read vaccine labels to determine which serovars are targeted, as immunity is believed to be serovar-specificPOTENTIAL IMPACTS OF DISEASE AGENT BEYOND CLINICAL ILLNESS Risks to public health●Leptospirosis is a zoonotic disease. Because clinical signs can be vague and maintenance hostscan be asymptomatic carriers, basic protective measures are suggested for at-risk populations (veterinarians, livestock owners, dairy workers, etc.): protect eyes with safety glasses, wear gloves especially if there are openings in the skin, thoroughly wash hands after interacting with animals of unknown status and before consuming food or water, etc.○Pregnant individuals within at-risk populations are particularly advised to utilise protective measures.●Many domestic animal species, including dogs, horses, and livestock, are susceptible to leptospirosisand could potentially transmit it to humans. Individuals should speak with local veterinarians to determine risk and appropriate prevention strategies, including animal vaccines.Risks to agriculture●If livestock or working animals (horses, dogs) develop clinical disease due to Leptospira, decreasedthrift and reproductive compromise can significantly impact production. Working animals may not beable to do their jobs as efficiently, and livestock may demand increased resources for treatment while producing less.REFERENCES AND OTHER INFORMATION●Atherstone, C., Picozzi, K., & Kalema-Zikusoka, G. (2014). Seroprevalence of Leptospira hardjo incattle and African buffalos in southwestern Uganda. The American Journal of Tropical Medicine and Hygiene, 90(2), 288–290.●Ayral, F., Djelouadji, Z., Raton, V., Zilber, A. L., Gasqui, P., et al. (2016). Hedgehogs and mustelidspecies: major carriers of pathogenic Leptospira, a survey in 28 animal species in France (20122015). PloS One, 11(9), e0162549.●Biscola, N. P., Fornazari, F., Saad, E., Richini-Pereira, V. B., Campagner, M. V., et al. (2011).Serological investigation and PCR in detection of pathogenic leptospires in snakes. Pesquisa Veterinária Brasileira, 31(9), 806-811.●Buhnerkempe, M. G., Pragger, K. C., Strelloff, C. C., Greig, D. J., Laake, J. L., et al. (2017). Detectingsignals of chronic shedding to explain pathogen persistence: Leptospira interrogans in California sea lions. Journal of Animal Ecology, 86(3), 460-472.●Denkinger, J., Guevara, N., Ayala, S., Murillo, J. C., Hirschfeld, M., et al. (2017). Pup mortality andevidence for pathogen exposure in Galapagos sea lions (Zalophus wollebaeki) on San Cristobal Island, Galapagos, Ecuador. Journal of Wildlife Disease, 53(3), 491-498.●Gravekamp, C., Korver, H., Montgomery, J., Everard, C. O. R., Carrington, D., et al. (1991).Leptospires isolated from toads and frogs on the island of Barbados. Zentralblatt für Bakteriologie, 275(3), 403-411.●Jobbins, S. E., Sanderson, S. E., & Alexander, K. A. (2014). Leptospira interrogans at the human-wildlife interface in northern Botswana: a newly identified public health threat. Zoonoses and Public Health, 61, 113-123.●Karesh, W. B., Hart, J. A., Hart, T. B., House, C., Torres, A., et al. (1995). Health evaluation of fivesympatric duiker species (Cephalophus spp). Journal of Zoo and Wildlife Medicine, 26(4), 485-502.●Leighton, F. A. & Kuiken, T. (2001). Leptospirosis. In E. S. Williams and I. K. Barker (Eds.), InfectiousDiseases of Wild Mammals (3rd ed., pp. 498-502). Iowa State Press.●Loffler, G. S., Rago, V., Martinez, M., Uhart, M., Florin-Christensen, M., et al. (2015). Isolation of aseawater tolerant Leptospira spp. from a southern right whale (Eubalaena australis). PLoS One, 10(12), e0144974.●Lunn, K. F. (2018). Overview of leptospirosis. Merck Veterinary Manual.Accessed 2020:https:///generalized-conditions/leptospirosis/overview-of-leptospirosis?query=leptospira●Pedersen, K., Anderson, T. D., Maison, R. M., Wiscomb, G. W., Pipas, M. J., et al. (2018). Leptospiraantibodies detected in wildlife in the USA and the US Virgin Islands. Journal of Wildlife Diseases, 54(3), 450-459.●Spickler, A. R. & Leedom, L. K. R. (2013). Leptospirosis. Accessed 2020:/Factsheets/pdfs/leptospirosis.pdf●The World Organisation for Animal Health (2018). Leptospirosis. Accessed 2020:https://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/3.01.12_LEPTO.pdf●Vieira, A. S., Pinto, P. S, & Lillenbaum, W. (2018). A systematic review of leptospirosis on wildanimals in Latin America. Tropical Animal Health and Production, 50(2), 229-238.●Wildlife Health Australia (2018). Leptospira infection in Australian mammals. Accessed 2020:https://.au/FactSheets.aspx●Wildlife Health Australia (2011). Leptospira infection in Australian seals. Accessed 2020:https://.au/FactSheets.aspx** *。

海洋微生物活性物质

海洋微生物活性物质的研究进展专业：生物工程姓名：李振森学号：4012010302海洋是生命的发源地，约占地球表面积的71%，其中生物种类20多万种，其多样性远远超过陆地生物的多样性。

由于海洋环境具有高盐度、高压、低营养、低温和无光照等条件，从而形成了海洋生物与陆地生物不同的生长方式和代谢系统。

近年来，随着人们对海洋生物研究的不断深入，发现了多种多样的生物及许多具有新颖、特异化学结构的生物活性物质。

海洋生物活性物质主要包括生物信息物质、生理活性物质、海洋生物毒素及生物功能材料等。

目前，从海洋生物中已相继发现300余种新型化合物，结构新颖并具有多样性：有枯类、聚醚类、当醇类、皂昔类、生物碱、多糖、小分子肤、核酸及蛋白质等，并具有丰富的生理及药理活性，包括抗菌、抗肿瘤、抗病毒、防治心血管疾病、延缓衰老及免疫调节等多种功能。

多年来，国内外一直致力于这方面的研究，试图从中开发结构明确，疗效肯定的新型生物活性物质，以用于攻克人类面临的重大疑难疾病，其中具有高生物活性和高选择性的海洋生物毒素备受重视，成为研究的热点。

近年来，海洋生物毒素是海洋生物活性物。

1、海洋抗肿瘤活性物质1.1海洋放线菌海洋有着极其丰富的放线菌资源，具有抗菌活性的海洋微生物中约有45%来源于放线菌。

就目前的报道，海洋放线菌产生的活性物质大部分来源于小单孢菌属和链霉菌属。

由于海洋放线菌所产生的代谢产物具有功能独特、结构新颖等特点而受到人们的广泛关注，例如抗真菌、抗疟等功能。

另一方面，陆生放线菌的不断开发，发现新的活性物质的可能性越发减少，迫使人们将目光转向海洋放线菌的开发。

1991年Fenical小组［1］首次发现一属全新的需盐生长的特殊海洋放线菌Salinispora，其广泛存在于热带和亚热带海泥中。

2003～2005 年Fenical小组从菌株Salinispora tropica CNB-392 中分离得到10个结构新颖的化合物［2－4］，其中化合物Salinosporamide A(1)［3］具有广阔的成药前景，对人结肠癌细胞的IC50为0．035 nmol /L，已作为癌症药物进入临床前研究［5－6］。

胰腺囊性病变的影像表现与临床特点(下)

国际医学放射学杂志ＩｎｔＪＭｅｄＲａｄｉｏｌ２０20Nov 鸦43穴6雪胰腺囊性病变的影像表现与临床特点（下）徐建国1唐光健2彭泰松1赵丽丽1于萍1任龙飞1许志高1【摘要】胰腺囊性病变（ＰＣＬ）是胰腺上皮和间质组织发生囊腔病变的一大类疾病，以胰腺内囊性包块为主要特征，具有不同的病因、临床和组织病理学特点。

本文的前两部分介绍了胰腺炎症相关囊性病变（包括胰腺假性囊肿与胰腺包裹性坏死）与胰腺真性囊肿（包括孤立性胰腺上皮囊肿、von Hippel-Lindau 病、多囊肾和囊性纤维化）以及常见的胰腺浆液性囊腺瘤、胰腺黏液性囊腺瘤和胰腺实性假乳头状瘤的影像表现与临床特点。

本篇为最后一部分，就常见的胰腺导管内乳头状黏液瘤与胰腺少见囊性肿瘤予以介绍和分析，以期为临床诊断与治疗提供重要依据。

【关键词】胰腺囊性病变；体层摄影术，X 线计算机；磁共振成像；导管内乳头状黏液瘤；囊性肿瘤中图分类号：R576；R445.2；R445.3文献标志码：APancreatic cystic lesions:imaging findings and clinical features (Part 3)XU Jianguo 1,TANG Guangjian 2,PENGTaisong 1,ZHAO Lili 1,YU Ping 1,REN Longfei 1,XU Zhigao 1.1Department of Radiology,Third People ’s Hospital of Datong City,Datong 037008,China;2Department of Radiology,Peking University First Hospital【Abstract 】Pancreatic cystic lesions (PCLs)are a broad spectrum of diseases with cystic lesions in pancreaticepithelial and interstitial tissues,characterized by cystic mass,with various etiology,clinical and histopathological characteristics.The first two parts of the article introduces the imaging manifestations and clinical features of 1)pancreatitis related cystic lesionincluding pancreatic pseudocyst and pancreatic encapsulated necrosis,2)pancreatic true cysts including isolated pancreatic epithelial cyst,von Hippel Lindau disease,polycystic kidney and cystic fibrosis,and3)the common cystic tumors of the pancreas including serous cystadenoma,mucinous cystadenoma and solid pseudopapilloma of the pancreas.In the last part of this issue,we introduce another common cystic tumor of pancreas (intraductal papillary mucinous neoplasm of the pancreas)and rare cystic tumors of the pancreas,in order to provide an important basis for clinical diagnosis and treatment of pancreatic cystic lesions.【Keywords 】Cystic disease of pancreas;Tomography,X -ray computed;Magnetic resonance imaging;Intraductalpapillary mucinous neoplasm;Cystic tumorＩｎｔＪＭｅｄＲａｄｉｏｌ，２０20，43（6）：716-720作者单位：1大同市第三人民医院医学影像科，大同037008；2北京大学第一医院放射科通信作者：唐光健，E-mail:***************.com DOI:10.19300/j.2020.J18513图文讲座3.4胰腺导管内乳头状黏液瘤胰腺导管内乳头状黏液瘤（intraductal papillary mucinous neoplasm,IPMN ）实际上并不是囊性肿瘤，由于肿瘤分泌黏液，引起胰腺导管扩张，大体病理与影像表现为伴有囊的病变，故在胰腺囊性病变内一并讨论。

新生儿败血症

病因及发病机制(Etiology and pathogenesis)
3、特异性免疫功能(Specific immune function)：
⑴新生儿体内IgG主要来自母体，且与胎龄相关，胎龄越小，IgG含量越低，因此早产儿更容易感染(IgG in neonatal mainly comes from the mother and is related to the gestational age. The smaller gestational age is, the lower IgG content is, so premature infants are more susceptible to infection)。
⑶由于未曾接触特异性抗原，T细胞为初始T细胞，产生细胞因子的能力低下，不能有效辅助B细胞、巨噬细胞、自然杀伤细胞和其他细胞参与免疫反应 (Since they have not been exposed to specific antigens, T cells are primary T cells with a low ability to produce cytokines and cannot effectively assist B cells, macrophages, natural killer cells and other cells in the immune response)。
⑸单核细胞产生粒细胞-集落刺激因子(G-CSF)、白细胞介素8(IL-8)等细胞因子的能力低下(The ability of monocytes to produce granulocyte-colony stimulating factor (g-csf), interleukin 8(il-8) and other cytokines is low)。