当前位置：文档库 › Trafficking of peripheral blood CD56bright cells to the

Trafficking of peripheral blood CD56bright cells to the

Journal of Reproductive Immunology

67(2005)21–34

Review

Traf?cking of peripheral blood CD56bright cells to the decidualizing uterus—new tricks for old dogmas?Marianne van den Heuvel a ,?,Crystal Peralta b ,Siamak Bashar c ,

Suzanne Taylor c ,Julie Horrocks d ,B.Anne Croy e

Department of Pediatrics,University of Western Ontario,Child Health Research Institute,London,Ont.,Canada N6C 2V5b Department of Anatomy and Cell Biology,University of Western Ontario,London,Ont.,Canada

c Department of Biomedical Sciences,University of Guelph,Ont.,Canada

d Department of Mathematics and Statistics,University of Guelph,Ont.,Canada

e Department o

f Anatomy and Cell Biology,Queen’s University,Kingston,Ont.,Canada

Received 11January 2005;received in revised form 21February 2005;accepted 16March 2005Abstract

CD56bright lymphocytes become abundant in the human uterus during every menstrual cycle,following the surge in pituitary-derived luteinizing hormone (LH),which initiates ?nal oocyte maturation.While the uterus is host to some CD56bright cells prior to ovulation,the rapid increase is thought to be due to proliferation of the resident population,accompanied by recruitment of CD56bright lymphocytes from the circulation.The rapid increase in CD56bright cells is concurrent with the onset of decidualization,the transformation of uterine stromal cells into secretory decidual cells.Uterine CD56bright cells proliferate and differentiate to become the predominant lymphocytes of the post-ovulatory uterus.These distinct,tissue-speci?c natural killer (NK)cells either die prior to menses or increase in number during early pregnancy,and then decline toward the end of the ?rst trimester.Since lymphocytes home to tissues from the circulation,we investigated mechanisms of NK

Abbreviations:A2M,alpha 2macroglobulin;dNK,decidual NK cell (human);DC,dendritic cell;E 2,estradiol;ER,estradiol receptor;HEV ,high endothelial venule;ICAM-1,intercellular adhesion molecule-1;LN,lymph node;LFA-1,leukocyte function associated molecule-1;LH,luteinizing hormone;LHR,luteinizing hormone receptor;LIF,leukemia inhibitory factor;MAdCAM,mucosal addressin cell adhesion molecule;NK,natural killer cell;PBL,peripheral blood lymphocytes;PNAd,peripheral lymph node addressin;PP,Peyers patches;P 4,progesterone;PR,progesterone receptor;PZP,pregnancy zone protein;SGA,small for gestational age;uNK,uterine NK cell (murine)?Corresponding author.Tel.:+15196858500x50070;fax:+15196858186.

E-mail address:mvandenh@uwo.ca (M.van den Heuvel).

doi:10.1016/j.jri.2005.03.004

22M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

cell traf?c over the course of natural menstrual cycles by measuring functional interactions between CD56+cells from blood and endothelial cells using the Stamper–Woodruff assay of lymphocyte adhesion to frozen tissue sections.While a baseline level of adhesion was maintained throughout the cycle,elevated l-selectin-dependent adhesion of peripheral blood CD56bright cells occurred during a peri-ovulatory window.However,there were no signi?cant menstrual cycle-induced changes in the transcription of l-selectin,alpha4integrin or LFA-1,or in expression of these proteins by NK cells,suggesting that the enhanced adhesion was due to post-translational modi?cations of these molecules.Quantitative RT-PCR failed to amplify the message for LH receptor or the alpha or beta forms of progesterone or estrogen receptors from blood NK cell subsets.Thus,we conclude that the actions of LH,E2,and P4on NK cells that promote interactions with endothelium and potential uterine homing are indirectly mediated through the responsiveness of other cell types.

Keywords:Decidualizing uterus;NK cell traf?cking;Peripheral blood CD56bright cells

1.Introduction

Human and murine endometria respond to ovarian steroid priming by transforming into mature decidua(Maslar,1988),a step essential for successful implantation(Genbacev et al.,2003).Amongst the many molecules secreted by decidua,speci?c cytokine and chemo-tactic signals and protease inhibitors are thought to orchestrate maternal contributions to successful implantation and the uterine modi?cations required to sustain pregnancy(Hanna et al.,2003;Kitaya et al.,2003b;Kao et al.,2002,2003).Due to dif?culties associated with the study of the peri-implantation uterus,the maternal compartment remains more enig-matic than fetal and placental development,but expression pro?ling studies using speci?c endometrial biopsies from fertile and infertile women during their menstrual cycle have recently become available and are likely to provide new insights into decidualization and implantation(Kao et al.,2002,2003;Carson et al.,2002;Borthwick et al.,2003).

Decidualization of human and murine endometria is associated with the appearance and proliferation of two unique maternal cell types:natural killer(NK)cells and immature dendritic cells(DC;King,2000;Chaouat et al.,2003;Kammerer et al.,2003;Robertson et al.,2003).These cells emerge in precise microdomains within implantation sites,proliferate and many localize to the spiral arteries.Studies of mice genetically-de?cient in lymphocytes identi?ed three uterine(u)NK cell functions:

(1)Support of terminal decidual cell differentiation.

(2)Sensitization of spiral arteries,which allows their pregnancy-associated dilation and

elongation.

(3)Formation of a transient lymphoid aggregate at the portal for vessels and nerves ser-

vicing implantation sites(Guimond et al.,1997;Croy et al.,2000).

These uNK cell-de?cient mice retain their fertility,delivering a normal number of pups. UNK cells mediate these functions through the secretion of interferon-?(IFN-?;Ashkar and Croy,1999;Ashkar et al.,2000),which changes expression of target genes,such as A2M,a major plasma cytokine binding and protease-inhibiting molecule active in spiral artery modi?cation(Croy et al.,2003;He et al.,2005).

M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–3423 In humans,decidua-associated(d)NK cells are present in low numbers in the pre-ovulatory uterus,but increase rapidly post-ovulation(King et al.,1996b).DNK cells are distinguished from peripheral blood NK cells by their expression of CD56at ten-fold higher levels,lack of expression of CD16and limited lytic ability(Moffett-King,2002).Despite the fact that timing of the increased abundance of dNK cells is tightly regulated by hor-mones,the hormone effects are thought to be indirect as dNK do not express progesterone receptors(PR)(King et al.,1996a)or the predominant estrogen receptor(ER?).DNK cells are reported to express ER?(Henderson et al.,2003),a form of ER that attenuates gene transcription(Kuiper and Gustafsson,1997).Indeed,dNK cells do not respond to hormonal stimulation in vitro by proliferation,cytokine secretion or alterations in lytic ability(King et al.,1996b;Kitaya et al.,2003a).

Progesterone regulates decidual production of cytokines,such as IL-15,necessary for NK cell proliferation and differentiation(Verma et al.,2000;Okada et al.,2000),leukemia inhibitory factor(LIF),required for implantation,and IL-11,which contributes to further decidualization in an autocrine manner(Robb et al.,2002;Dimitriadis et al.,2002).Proges-terone also induces stromal cell production of CXCL9(Mig)and CXCL10(IP-10;Kitaya et al.,2004),ligands of CXCR3,a chemokine receptor expressed by peripheral blood NK cells(Campbell et al.,2001;Inngjerdingen et al.,2001).Chemokine receptors that are highly up-regulated in decidua during the luteal phase include CXCR1,CCR2b,and CCR5 (Dominguez et al.,2003).Thus,the major roles of P4include induction of decidualization, induction of chemokines capable of recruiting NK cells,and up-regulation of IL-15,the cytokine essential for differentiation and maintenance of dNK cells(Ashkar et al.,2003).

In contrast,the ovarian hormone estradiol(E2)induces vasodilation or relaxation in vessels through interactions with endothelial cells throughout the cardiovascular system (Sullivan et al.,1995;Mendelsohn,2002).These effects are particularly profound in the pregnant uterus,where branches from the uterine arteries known as spiral arteries,elongate, dilate,and become tortuous.Spiral artery modi?cation yields high-?ow,low-resistance vessels with reduced wall structure and proximal(i.e.toward the fetus)replacement of endothelium with intravascular trophoblast.These structural changes,which occur during the third and fourth months of human pregnancy,shortly after the onset of the decline in dNK cells and between gestation days9and10in mice,when uNK cells are at their peak in num-ber and function,are thought to be required in humans for normal rates of conceptus growth and maternal health in later gestation(Sattar et al.,2003).The cells controlling spiral artery modi?cation in mice and women appear to differ(uNK versus trophoblast cells,respec-tively).However,this difference may lie in the proportion of the vessel changed by each cell type(i.e.uNK cells functioning more distally to fetal trophoblasts and the spiral arterial seg-ment being relatively longer in mice)or trophoblast invasion in humans may be facilitated by prior dNK cell sensitization of these vessels(as occurs in mice)during the implantation period(Adamson et al.,2003).Human decidual(d)NK cells,like their murine counterparts, express major angiogenic molecules,such as vascular endothelial and placental growth factors(Smith,2001).Thus,if the roles of uNK cells in mice and dNK cells in humans are analogous,dNK support of decidualization can be linked to fertility/implantation,while dNK contributions to spiral artery modi?cation can be associated with the development of pre-eclampsia and some small-for-gestational-age(SGA)pregnancies(Khong et al., 1986).

24M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

2.Traf?cking of pre-dNK cells to the uterus

Self-renewing murine uNK cell progenitors do not reside within the uterus(Chantakru

et al.,2002).Transplantable progenitors of murine uNK cells were found in all lymphoid

tissues,but particularly in the spleen during pregnancy,suggesting a hormonal induction

of NK cell mobility or maturation(Chantakru et al.,2002).Mobilization of blood-borne

lymphocytes to tissue depends on their sequential adhesive interactions with endothelial

cells under wall shear stress induced by hemodynamic?ow(Springer,1995).

Homing of naive T and B lymphocytes to secondary lymphoid tissues,such as lymph

nodes(LN)and Peyer’s patches(PP)is a well-described four-step cascade of events that

occurs in post-capillary venules in secondary immune tissues or at sites of in?amma-

tion(Springer,1994;Butcher and Picker,1996;V on Andrian and Mempel,2003).First,a

sequence of transient adhesive interactions under shear force between adhesion molecules

constitutively expressed on circulating lymphocytes(l-selectin and/or?4?7integrin)and their respective ligands,peripheral node addressins(PNAd,includes CD34,GlyCAM-1,

podocalyxin,Sgp200;Rosen,2004)and mucosal addressin cell adhesion molecule(Mad-

CAM)on specialized high endothelial venules(HEV)cause circulating lymphocytes to

progressively slow,tether,roll,and stop.Subsequent rapid activation by chemokines,such

as CCL19(MIP3?),CCL21(6Ckine),and CXCL12(SDF1)induces lateral mobility and

temporarily enhanced af?nity of LFA-1for its ligand,ICAM-1,resulting in?rm attachment,

then transendothelial migration along a chemokine gradient into tissue.Endothelial expres-

sion of ligands and tissue-speci?c secretion of chemoattractants enable B cell migration at

follicular sites and T cell egress at interfollicular areas in LN or PP(Warnock et al.,2000;

Constantin et al.,2000;Butcher et al.,1999).

These steps in homing apply to extravasation of leukocytes to tertiary sites in response

to speci?c in?ammation-induced chemokine signals.Migrating cells include memory lym-

phocytes,dendritic cells,and neutrophils(Picker and Butcher,1992).Sites of in?ammation

are also characterized by production of cytokines,which up-regulate expression of adhe-

sion molecules,such as E-selectin,VCAM-1and ICAM-1on vascular endothelium that

facilitate egress of immune cells(Butcher et al.,1999).

Much less is known about NK cell traf?cking.Circulating NK cells are subdivided based

on expression of CD3,CD16,and CD56.The majority of NK cells express CD16strongly

and CD56weakly(CD56dim),but about10%of blood NK cells have higher expression of

CD56and lack expression of CD16(CD56bright;Robertson and Ritz,1990).CD3expressing

CD56+cells are designated NK-T cells because they also express a somatically rearranged ??T cell receptor,and are thus considered to be of T cell lineage rather than NK cell lineage. The CD56bright subset is further discriminated from the CD56dim subset by expression of high af?nity IL-2R,high expression of l-selectin and?4and?1integrins,but low expression of LFA-1(Frey et al.,1998).CD56bright and dim cells also differ in their biologic functions. CD56dim cells have greater lytic ability,while CD56bright cells have immunoregulatory roles through secretion of cytokines(Cooper et al.,2001).CD56+populations respond uniquely to chemotactic signaling;CD56dim cells migrate in response to CXCL8(IL-8)and CX3CL1 (fractalkine),and strongly express their receptors CXCR1and CX3CR1.CD56bright cells express the chemokine receptors CCR5,which binds CCL3(MIP?1),CCL4(MIP?1)and CCL5(RANTES),and CCR7,which binds CCL19(MIP?3)and CCL21(6Ckine;Campbell

M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–3425 Table1

Traf?cking phenotype of natural killer cell subsets

Blood subsets Decidual NK

Dim Bright NK-T

CD56+++++++

CD16+???

CD3??+?

l-Selectin++++?

Alpha4integrin++++++

LFA-1++++

CCR1??+++

CCR2??+++

CCR5?+++++

CCR7?++?+/?

CXCR1+???

CXCR2+?+?

CXCR3+++++++

CXCR4++++++++

CX3CR1+++++++

(+)Expressed;(++)strongly expressed;(+++)very strongly expressed;(?)not expressed.

et al.,2001).NK-T cells share expression of all chemokine receptors found on the CD56bright subset and have in addition CCR1,2,and6.While all CD56+populations express CXCR3-binds CXCL9(Mig),CXCL10(IP-10),and CXCL11(ITAC)-and CXCR4-binds CXCL12 (SDF1)-NK cells do not readily migrate to secondary lymphoid tissues,such as LN or PP, but are found in spleen(Campbell et al.,1998).Decidual NK cells have a unique expression pattern,strongly expressing,CCR1,2,and5,CXCR3and4,and CX3CR1(Red-Horse et al.,2001).The expression of both phenotypic and chemokine receptors for each CD56+ population is summarized in Table1.CD56bright cells,through their intense expression of l-selectin(Frey et al.,1998;Kruse et al.,1999)and speci?c chemokine receptor array (Campbell et al.,2001;Inngjerdingen et al.,2001)and their ability to adhere in an l-selectin-dependent manner to frozen sections of LN(Frey et al.,1998),appear predisposed to tissue-selective homing.We addressed whether l-selectin-mediated interactions could contribute to the traf?cking of NK cells or their precursors to the decidualizing uterus.

The Stamper–Woodruff adhesion assay is a model of lymphocyte traf?cking poten-tial(Stamper and Woodruff,1976)that evaluates speci?c functional adhesive interactions between viable lymphocytes and endothelium presented in frozen tissue sections.The assay is performed on a rotating table to mimic the shear forces of?owing blood nec-essary to engage adhesion molecules on both lymphocytes and https://www.wendangku.net/doc/8c6809169.html,ing this assay,we demonstrated that lymphocytes from pregnant or ovarian steroid hormone-treated ovariectomized mice had increased interactions with endothelium over lymphocytes from non-pregnant donors(Chantakru et al.,2003).These interactions were mediated by two well-characterized lymphocyte homing receptors,l-selectin and alpha4integrin,since pre-treatment of the lymphocytes with function-blocking antibodies to either of the molecules, or the substrate tissue with antibodies to their counter-receptors PNAd and MAdCAM, removed the enhancement.Furthermore,independent but coordinated,pregnancy and

26M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

steroid hormone-induced changes occurred in endothelium of the uterus and LN but not in endothelium of other organs.The changes in endothelia were recognized by murine splenocytes,human blood lymphocytes,and indicator cell lines expressing speci?c recep-tors(Chantakru et al.,2003).Importantly,we found that the human lymphocytes adhering to microvessels of midgestation decidua were enriched with CD56bright NK cells.While CD56bright NK cells made up～3%of the blood lymphocytes applied to the sections,they constituted75%of the adherent cells.

3.Recruitment of decidual natural killer cells

Because stromal triggering toward decidualization and uterine recruitment of NK cells coincide in women during each menstrual cycle,we hypothesized that CD56bright cells in human blood contained precursors of dNK cells and that interactions of these cells with uter-ine endothelium would vary over the menstrual cycle.We further postulated that endocrine changes in the menstrual cycle induce synchronous changes in uterine endothelium that pro-mote NK cell traf?cking into the human uterus and that defects in the migration pathway or its regulation could compromise blastocyst implantation and/or limit structural changes to the arteries supporting human placental development.Because the substrate tissue needed for the in vitro cell adhesion assay has a limited useful storage life of10–15days,uterine tissue from inbred mice was used as the substrate tissue for evaluating endocrinological modi?cations of the functions of human blood NK cells.

Serial blood samples from18women with normal menstrual cycles were studied.NK cell subsets,hormone levels,and functional adhesion to uterine endothelium from gd7mice were monitored over a menstrual cycle.The adhesive function of total peripheral blood lymphocytes(PBL)and the CD56bright cell subset were studied on alternate days using endothelium in LN(n=14)or decidualized uterus(n=6).Fig.1depicts the highlights of this study.On LN substrates,adhesion of PBL and CD56bright cells was highly variable between donors,but consistently peaked at the luteinizing hormone(LH)surge.This rise in adhesion,however,did not reach statistical signi?cance.With uterine endothelial substrates, dramatic gains in adhesion were observed for both PBL and NK cells(p=0.004)with all adhesion localized to decidua basalis(Van den Heuvel et al.,2005).

The strongest statistical correlation was between estrogen concentration and number of adherent cells.During a typical28-day menstrual cycle,E2levels rise from cycle day8to cycle day13,when a surge of LH down-regulates E2.We found that CD56bright adhesion peaked only during a narrow window between cycle days10and12,just prior to ovulation (cycle day14).Thus,it appeared that traf?cking of peripheral blood NK cells might be directly regulated by rising levels of E2and/or LH,but not P4.Function blocking antibodies to l-selectin or alpha4integrin signi?cantly reduced adhesion and eliminated the peri-ovulatory enhancement,suggesting that recruitment is dependent upon classic adhesion molecules used in in?ammatory processes.

To further investigate the role of hormones in the adhesive interactions of NK cells with endothelium,total lymphocytes collected from male donors,who would not be primed by menstrual stage,were cultured in serum-free Roswell Park Memorial Institute medium (RPMI)with LH,E2,and P4.Hormone effects were titrated in ranges found in plasma during

M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–3427

Fig.1.Adhesion of peripheral blood lymphocytes(PBL)and CD56bright cells to mouse decidua.Mean adhesion of human PBL or CD56bright cells to frozen endothelium from mice at gd7a three time points during a menstrual cycle is presented.Five106cells were overlaid onto frozen tissue under shear for30min,and then non-adhesive cells were rinsed off.Adhesion was evaluated in50?elds by two researchers.Panels A and B show mean adhesion to high endothelial venules(HEV)in lymph nodes(LN),while C and D show adhesion to decidua.Panels A and C illustrate total PBL adhesion and B and D depict adhesion of CD56bright cells only.Solid bars show untreated cells, while open bars show adhesion of identical aliquots pre-treated with a function blocking antibody to l-selectin. the menstrual cycle.LH ranged from5to250IU/ml,E2from100to800pg/ml,and P4from 1to8ng/ml.After4h in culture,signi?cant changes occurred in CD56cell adhesion to endothelium from gd seven mouse decidua basalis.These experiments are summarized in Fig.2.A positive association of adhesion was found with E2and with LH,but not with P4for both PBL and CD56bright cells.The effects of E2and LH were not additive when combined treatment was used.These data strongly suggest hormonal complicity in the peri-ovulatory change in the functional adhesion of NK cells.

In further studies,quantitative PCR was performed on NK cell subsets derived from magnetically separated PBL of women at day5or at day of LH surge.No transcripts for either the ER or PR isoforms,or for the LH receptor were detected in any of the subsets, although GAPDH and genes for l-selectin,alpha4integrin,and LFA-1were detected and the hormone receptor transcripts were ampli?ed from control tissues(ovary or uterus) as summarized in Table2.Thus,the hormonally effected changes in NK cell function must be mediated through another cell population in the PBL cultures.In-depth studies of direct endocrinological involvement in maternal immune regulation have often given apparently con?icting results.Effects of hormones,especially E2,on NK cell numbers, subsets,and activity have been extensively reported in relation to the menstrual cycle(Hunt

28M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

Fig.2.The role of hormones in the adhesive interactions of NK cells with endothelium.Total lymphocytes collected from three male donors were cultured for4h in serum-free RPMI alone or with varying concentrations of estradiol(E2;panel A),luteinizing hormone(LH;panel B)or progesterone(P4;panel C)as shown on the lower axis,then applied to frozen sections of gestational mouse decidua.Post-culture adhesion of CD56bright cells from individual donors to mouse decidua is shown.

M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–3429 Table2

Summary of real-time RT-PCR analyses for genes transcribed in blood NK cell subsets

Dim Bright NK-T Ovary/uterus

d5LH d5LH d5LH

LHR??????+

ER???????+

PR A??????+

PR B??????+

L-Selectin748484ND

Alpha4integrin12591251871124511151120ND

LFA-1211412151110ND GAPDH+++++++

Values are copy numbers relative to the average GAPDH expression from seven samples:(+)positive;(?)not detected;ND:not done.

Fig.3.A model of pre-dNK cell traf?cking from lymphoid reservoirs to decidualizing uterus.In the early secretory phase and luteal phase of the menstrual cycle,few natural killer cells(?)enter and leave the uterus in normal immune surveillance.Increased peri-ovulatory plasma concentrations of E2and/or LH induce expression of unknown factors(),which mobilize NK cell egress from reservoirs into the circulation,particularly LN. NK cells leave the blood in increased numbers and preferentially enter the uterus in response to a gradient of chemokines(CXCR3and4ligands())secreted by the E 2-primed stromal cells in the post-ovulatory uterus. l-Selectin is shed as they traverse the decidua and localize in decidual cells proximal to blood vessels.P4acts on E2-primed stroma to initiate decidualization as well as on vascular endothelium to promote NK cell-secreted IFN?(thinning of the vessel wall and dilation of the major arteries(Kitaya et al.,2004;Red-Horse et al.,2001; Hanna et al.,2003).

30M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

et al.,2000;Flynn et al.,2000;Bouman et al.,2001;Yovel et al.,2001;Sulke et al.,1985; Souza et al.,2001;Jones et al.,1997),pregnancy(Veenstra et al.,2002;Watanabe et al., 1997),reproductive pathologies(Provinciali et al.,1995;Somigliana et al.,1999;Searle et al.,1999),contraceptive use(Auerbach et al.,2002;Scanlan et al.,1995),and metastatic disease(Baral et al.,1995;Roszkowski et al.,1997),but there are no reports of an in?uence of sex hormones on NK cell traf?cking in the human.

Quantitative PCR was also undertaken on the above samples to determine whether changes in transcription of adhesion molecules might account for enhanced lymphocyte interactions with endothelium at the LH surge.There were no differences in the numbers of transcripts for l-selectin,alpha4integrin,or LFA-1between day5and day of LH surge in either of the NK cell subsets.This con?rmed our earlier observation by?ow cytometry that there was no change in mean?uorescence intensity in expression of l-selectin,alpha4 integrin or LFA-1over the menstrual cycle.These results suggest that hormone-promoted changes in membrane receptor clustering must be investigated.

4.Conclusions

The changes in lymphocyte/endothelial interactions that we have described have the potential to underlie cyclic movement of progenitors of dNK cells from peripheral lymphoid tissue to the uterus.Once localized to peri-vascular and decidual sites where they are in contact with fetal trophoblast,dNK cells contribute to modifying the endometrial environment to promote implantation and alter the maternal vasculature.These inaugural studies demonstrate dynamic,coordinated,indirect regulation of adhesive molecule function on CD56bright cells and endothelium by ovarian steroid hormones,as summarized in Fig.3.

Acknowledgements

The work described here was supported by NSERC,CIHR,the Ontario Women’s Health Scholarship Award and the Canada Research Chairs Program.We thank J.E.Lewis,A. Simpson,K.Hatta,M.E.Junkins,S.Burke,and Y.Fang for technical support and Dr.C. Tayade for his most helpful discussions.We greatly appreciate the co-operation of our blood donors without whom our study would not have been possible.

References

Adamson,S.L.,Lu,Y.,Whiteley,K.J.,Holmyard,D.,Hemberger,M.,Pfarrer,C.,Cross,J.C.,2003.Interactions between trophoblast cells and the maternal and fetal circulation in the mouse placenta.Dev.Biol.250,358–373.

Ashkar,A.A.,Croy,B.A.,1999.Interferon-gamma contributes to the normalcy of murine pregnancy.Biol.Reprod.

61,493–502.

Ashkar,A.A.,di Santo,J.P.,Croy,B.A.,2000.Interferon gamma contributes to initiation of uterine vascular modi?cation,decidual integrity,and uterine natural killer cell maturation during normal murine pregnancy.J.

Exp.Med.192,259–270.

M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–3431 Ashkar,A.A.,Black,G.P.,Wei,Q.,He,H.,Liang,L.,Head,J.R.,Croy,B.A.,2003.Assessment of requirements for IL-15and IFN regulatory factors in uterine NK cell differentiation and function during pregnancy.J.Immunol.

171,2937–2944.

Auerbach,L.,Hafner,T.,Huber,J.C.,Panzer,S.,2002.In?uence of low-dose oral contraception on peripheral blood lymphocyte subsets at particular phases of the hormonal cycle.Fertil.Steril.78,83–89.

Baral,E.,Nagy,E.,Berczi,I.,1995.Modulation of natural killer cell-mediated cytotoxicity by tamoxifen and estradiol.Cancer75,591–599.

Borthwick,J.M.,Charnock-Jones,D.S.,Tom,B.D.,Hull,M.L.,Teirney,R.,Phillips,S.C.,Smith,S.K.,2003.

Determination of the transcript pro?le of human endometrium.Mol.Hum.Reprod.9,19–33.

Bouman,A.,Moes,H.,Heineman,M.J.,de Leij,L.F.,Faas,M.M.,2001.Cytokine production by natural killer lymphocytes in follicular and luteal phase of the ovarian cycle in humans.Am.J.Reprod.Immunol.45, 130–134.

Butcher,E.C.,Picker,L.J.,1996.Lymphocyte homing and homeostasis.Science272,60–66.

Butcher,E.C.,Williams,M.,Youngman,K.,Rott,L.,Briskin,M.,1999.Lymphocyte traf?cking and regional immunity.Adv.Immunol.72,209–253.

Campbell,J.J.,Bowman,E.P.,Murphy,K.,Youngman,K.R.,Siani,M.A.,Thompson,D.A.,Wu,L.,Zlotnik,

A.,Butcher,E.C.,1998.6-C-kine(SLC),a lymphocyte adhesion-triggering chemokine expressed by high

endothelium,is an agonist for the MIP-3beta receptor CCR7.J.Cell Biol.141,1053–1059.

Campbell,J.J.,Qin,S.,Unutmaz,D.,Soler,D.,Murphy,K.E.,Hodge,M.R.,Wu,L.,Butcher,E.C.,2001.Unique subpopulations of CD56+NK and NK-T peripheral blood lymphocytes identi?ed by chemokine receptor expression repertoire.J.Immunol.166,6477–6482.

Carson,D.D.,Lagow,E.,Thathiah,A.,Al-Shami,R.,Farach-Carson,M.C.,Vernon,M.,Yuan,L.,Fritz,M.A., Lessey,B.,2002.Changes in gene expression during the early to mid-luteal(receptive phase)transition in human endometrium detected by high-density microarray screening.Mol.Hum.Reprod.8,871–879. Chantakru,S.,Miller,C.,Roach,L.E.,Kuziel,W.A.,Maeda,N.,Wang,W.C.,Evans,S.S.,Croy,B.A.,2002.Con-tributions from self-renewal and traf?cking to the uterine NK cell population of early pregnancy.J.Immunol.

168,22–28.

Chantakru,S.,Wang,W.C.,van den Heuvel,M.,Bashar,S.,Simpson,A.,Chen,Q.,Croy,B.A.,Evans,S.S.,2003.

Coordinate regulation of lymphocyte-endothelial interactions by pregnancy-associated hormones.J.Immunol.

171,1132–1145.

Chaouat,G.,Ledee-Bataille,N.,Zourbas,S.,Dubanchet,S.,Sandra,O.,Martal,J.,Ostojojic,S.,Frydman,R., 2003.Implantation:can immunological parameters of implantation failure be of interest for pre-eclampsia?J.

Reprod.Immunol.59,205–217.

Constantin,G.,Majeed,M.,Giagulli,C.,Piccio,L.,Kim,J.Y.,Butcher,E.C.,Laudanna,C.,2000.Chemokines trigger immediate beta2integrin af?nity and mobility changes:differential regulation and roles in lymphocyte arrest under?ow.Immunity13,759–769.

Cooper,M.A.,Fehniger,T.A.,Turner,S.C.,Chen,K.S.,Ghaheri,B.A.,Ghayur,T.,Carson,W.E.,Caligiuri,M.A., 2001.Human natural killer cells:a unique innate immunoregulatory role for the CD56(bright)subset.Blood 97,3146–3151.

Croy,B.A.,di Santo,J.P.,Greenwood,J.D.,Chantakru,S.,Ashkar,A.A.,2000.Transplantation into genetically alymphoid mice as an approach to dissect the roles of uterine natural killer cells during pregnancy—a review.

Placenta21(Suppl.A),S77–S80.

Croy,B.A.,Esadeg,S.,Chantakru,S.,van den Heuvel,M.,Paffaro Jr.,V.A.,He,H.,Black,G.,Ashkar,A.A., Kiso,Y.,Zhang,J.,2003.Update on pathways regulating the activation of uterine natural killer cells,their interactions with decidual spiral arteries and homing of their precursors to the uterus.J.Reprod.Immunol.59, 175–191.

Dimitriadis,E.,Robb,L.,Salamonsen,L.A.,2002.Interleukin11advances progesterone-induced decidualization of human endometrial stromal cells.Mol.Hum.Reprod.8,636–643.

Dominguez,F.,Galan,A.,Martin,J.J.,Remohi,J.,Pellicer,A.,Simon,C.,2003.Hormonal and embryonic regulation of chemokine receptors CXCR1,CXCR4,CCR5and CCR2B in the human endometrium and the human blastocyst.Mol.Hum.Reprod.9,189–198.

Flynn,L.,Byrne,B.,Carton,J.,Kelehan,P.,O’Herlihy,C.,O’Farrelly,C.,2000.Menstrual cycle dependent?uc-tuations in NK and T-lymphocyte subsets from non-pregnant human endometrium.Am.J.Reprod.Immunol.

43,209–217.

32M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

Frey,M.,Packianathan,N.B.,Fehniger,T.A.,Ross,M.E.,Wang,W.C.,Stewart,C.C.,Caligiuri,M.A.,Evans, S.S.,1998.Differential expression and function of l-selectin on CD56bright and CD56dim natural killer cell subsets.J.Immunol.161,400–408.

Genbacev,O.D.,Prakobphol,A.,Foulk,R.A.,Krtolica,A.R.,Ilic,D.,Singer,M.S.,Yang,Z.Q.,Kiessling,L.L., Rosen,S.D.,Fisher,S.J.,2003.Trophoblast l-selectin-mediated adhesion at the maternal-fetal interface.Sci-ence299,405–408.

Guimond,M.J.,Luross,J.A.,Wang,B.,Terhorst,C.,Danial,S.,Croy,B.A.,1997.Absence of natural killer cells during murine pregnancy is associated with reproductive compromise in TgE26mice.Biol.Reprod.56, 169–179.

Hanna,J.,Wald,O.,Goldman-Wohl,D.,Prus,D.,Markel,G.,Gazit,R.,Katz,G.,Haimov-Kochman,R.,Fujii,N., Yagel,S.,Peled,A.,Mandelboim,O.,2003.CXCL12expression by invasive trophoblasts induces the speci?c migration of CD16-human natural killer cells.Blood102,1569–1577.

He,H.,McCartney,D.J.,Wei,Q.,Esadeg,S.M.,Zhang,J.,Foster,R.A.,Hayes,M.A.,Tayade,C.,Van Leuven,

F.,Croy,B.A.,2005.Characterization of a murine alpha2macroglobulin gene expressed in reproductive and

cardiovascular tissue.Biol.Reprod.72,266–275.

Henderson,T.A.,Saunders,P.T.,Moffett-King,A.,Groome,N.P.,Critchley,H.O.,2003.Steroid receptor expres-sion in uterine natural killer cells.J.Clin.Endocrinol.Metab.88,440–449.

Hunt,J.S.,Miller,L.,Roby,K.F.,King,A.,2000.Female steroid hormones regulate production of pro-in?ammatory molecules in uterine leukocytes.Hum.Reprod.Update6,28–36.

Inngjerdingen,M.,Damaj,B.,Maghazachi,A.A.,2001.Expression and regulation of chemokine receptors in human natural killer cells.Blood97,367–375.

Jones,R.K.,Bulmer,J.N.,Searle,R.F.,1997.Cytotoxic activity of endometrial granulated lymphocytes during the menstrual cycle in humans.Biol.Reprod.57,1217–1222.

Kammerer,U.,Eggert,A.O.,Kapp,M.,McLellan,A.D.,Geijtenbeek,T.B.,Dietl,J.,van Kooyk,Y.,Kampgen,

E.,2003.Unique appearance of proliferating antigen-presenting cells expressing DC-SIGN(CD209)in the

decidua of early human pregnancy.Am.J.Pathol.162,887–896.

Kao,L.C.,Tulac,S.,Lobo,S.,Imani,B.,Yang,J.P.,Germeyer,A.,Osteen,K.,Taylor,R.N.,Lessey,B.A.,Giudice, L.C.,2002.Global gene pro?ling in human endometrium during the window of implantation.Endocrinology 143,2119–2138.

Kao,L.C.,Germeyer,A.,Tulac,S.,Lobo,S.,Yang,J.P.,Taylor,R.N.,Osteen,K.,Lessey,B.A.,Giudice,L.C., 2003.Expression pro?ling of endometrium from women with endometriosis reveals candidate genes for disease-based implantation failure and infertility.Endocrinology144,2870–2881.

Khong,T.Y.,De Wolf,F.,Robertson,W.B.,Brosens,I.,1986.Inadequate maternal vascular response to placentation in pregnancies complicated by pre-eclampsia and by small-for-gestational age infants.Br.J.Obstet.Gynaecol.

93,1049–1059.

King,A.,2000.Uterine leukocytes and decidualization.Hum.Reprod.Update6,28–36.

King,A.,Gardner,L.,Loke,Y.W.,1996a.Evaluation of oestrogen and progesterone receptor expression in uterine mucosal lymphocytes.Hum.Reprod.11,1079–1082.

King,A.,Jokhi,P.P.,Burrows,T.D.,Gardner,L.,Sharkey,A.M.,Loke,Y.W.,1996b.Functions of human decidual NK cells.Am.J.Reprod.Immunol.35,258–260.

Kitaya,K.,Yasuda,J.,Nakayama,T.,Fushiki,S.,Honjo,H.,2003a.Effect of female sex steroids on human endometrial CD16neg CD56bright natural killer cells.Fertil.Steril.79(Suppl.1),730–734.

Kitaya,K.,Yasuda,J.,Yagi,I.,Tada,Y.,Fushiki,S.,Honjo,H.,2003b.IL-15expression at human endometrium and decidua.Biol.Reprod.63,683–687.

Kitaya,K.,Nakayama,T.,Daikoku,N.,Fushiki,S.,Honjo,H.,2004.Spatial and temporal expression of ligands for CXCR3and CXCR4in human endometrium.J.Clin.Endocrinol.Metab.89,2470–2476.

Kruse,A.,Merchant,M.J.,Hallmann,R.,Butcher,E.C.,1999.Evidence of specialized leukocyte-vascular homing interactions at the maternal/fetal interface.Eur.J.Immunol.29,1116–1126.

Kuiper,G.G.,Gustafsson,J.A.,1997.The novel estrogen receptor-beta subtype:potential role in the cell-and promoter-speci?c actions of estrogens and anti-estrogens.FEBS Lett.410,87–90.

Maslar,I.A.,1988.The progestational endometrium.Semin.Reprod.Immunol.6,115–128.

Mendelsohn,M.E.,2002.Genomic and nongenomic effects of estrogen in the vasculature.Am.J.Cardiol.90, 3F–6F.

Moffett-King,A.,2002.Natural killer cells and pregnancy.Nat.Rev.Immunol.2,656–663.

M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–3433 Okada,H.,Nakajima,T.,Sanezumi,M.,Ikuta,A.,Yasuda,K.,Kanzaki,H.,2000.Progesterone enhances interleukin-15production in human endometrial stromal cells in vitro.J.Clin.Endocrinol.Metab.85, 4765–4770.

Picker,L.J.,Butcher,E.C.,1992.Physiological and molecular mechanisms of lymphocyte homing.Annu.Rev.

Immunol.10,561–591.

Provinciali,M.,Di Stefano,G.,Muzzioli,M.,Garzetti,G.G.,Ciavattini,A.,Fabris,N.,1995.Relationship between 17-beta-estradiol and prolactin in the regulation of natural killer cell activity during progression of endometrio-sis.J.Endocrinol.Invest.18,645–652.

Red-Horse,K.,Drake,P.M.,Gunn,M.D.,Fisher,S.J.,2001.Chemokine ligand and receptor expression in the pregnant uterus:reciprocal patterns in complementary cell subsets suggest functional roles.Am.J.Pathol.

159,2199–2213.

Robb,L.,Dimitriadis,E.,Li,R.,Salamonsen,L.A.,2002.Leukemia inhibitory factor and interleukin-11:cytokines with key roles in implantation.J.Reprod.Immunol.57,129–141.

Robertson,M.J.,Ritz,J.,1990.Biology and clinical relevance of human natural killer cells.Blood76,2421–2438.

Robertson,S.A.,Redman,C.W.,McCracken,S.A.,Hunt,J.S.,Dimitriadis,E.,Moffett-King,A.,Chamley,L., 2003.Immune modulators of implantation and placental development—a workshop report.Placenta24,S16–S20.

Rosen,S.D.,2004.Ligands for l-selectin:homing,in?ammation,and beyond.Annu.Rev.Immunol.22,129–156. Roszkowski,P.I.,Hyc,A.,Stopinska-Gluszak,U.,Malejczyk,J.,1997.Natural killer cell activity and sex hormone levels in mastopathy.Gynecol.Endocrinol.11,399–404.

Sattar,N.,Ramsay,J.,Crawford,L.,Cheyne,H.,Greer,I.A.,2003.Classic and novel risk factor parameters in women with a history of preeclampsia.Hypertension42,39–42.

Scanlan,J.M.,Werner,J.J.,Legg,R.L.,Laudenslager,M.L.,1995.Natural killer cell activity is reduced in asso-ciation with oral contraceptive use.Psychoneuroendocrinology20,281–287.

Searle,R.F.,Jones,R.K.,Bulmer,J.N.,1999.Phenotypic analysis and proliferative responses of human endometrial granulated lymphocytes during the menstrual cycle.Biol.Reprod.60,871–878.

Smith,S.K.,2001.Regulation of angiogenesis in the endometrium.Trends Endocrinol.Metab.12,147–151. Somigliana,E.,Vigano,P.,Vignali,M.,1999.Endometriosis and unexplained recurrent spontaneous abortion: pathological states resulting from aberrant modulation of natural killer cell function?Hum.Reprod.Update 5,40–51.

Souza,S.S.,Castro,F.A.,Mendonca,H.C.,Palma,P.V.,Morais,F.R.,Ferriani,R.A.,V oltarelli,J.C.,2001.In?uence of menstrual cycle on NK activity.J.Reprod.Immunol.50,151–159.

Springer,T.A.,1994.Traf?c signals for lymphocyte recirculation and leukocyte emigration:the multistep paradigm.

Cell76,301–314.

Springer,T.A.,1995.Traf?c signals on endothelium for lymphocyte recirculation and leukocyte emigration.Annu.

Rev.Physiol.57,827–872.

Stamper,H.B.J.,Woodruff,J.J.,1976.Lymphocyte homing into lymph nodes:in vitro demonstration of the selective af?nity of recirculating lymphocytes for high-endothelial venules.J.Exp.Med.144,828–833. Sulke,A.N.,Jones,D.B.,Wood,P.J.,1985.Variation in natural killer activity in peripheral blood during the menstrual cycle.Br.Med.J.(Clin.Res.Ed.)290,884–886.

Sullivan,T.R.J.,Karas,R.H.,Aronovitz,M.,Faller,G.T.,Ziar,J.P.,Smith,J.J.,O’Donnell,T.F.J.,Mendelsohn, M.E.,1995.Estrogen inhibits the response-to-injury in a mouse carotid artery model.J.Clin.Invest.96, 2482–2488.

Van den Heuvel,M.,Horrocks,J.,Bashar,S.,Taylor,S.,Burke,S.,Hatta,K.,Lewis,J.E.,Croy,B.A.,2005.Men-strual cycle hormones induce changes in functional interactions between lymphocytes and decidual vascular endothelial cells.J.Clin.Endocrinol.Metab.90,2835–2842.

Veenstra van Nieuwenhoven,A.L.,Bouman,A.,Moes,H.,Heineman,M.J.,de Leij,L.F.,Santema,J.,Faas,M.M., 2002.Cytokine production in natural killer cells and lymphocytes in pregnant women compared with women in the follicular phase of the ovarian cycle.Fertil.Steril.77,1032–1037.

Verma,S.,Hiby,S.E.,Loke,Y.W.,King,A.,2000.Human decidual natural killer cells express the receptor for and respond to the cytokine interleukin15.Biol.Reprod.62,959–968.

V on Andrian,U.H.,Mempel,T.R.,2003.Homing and cellular traf?c in lymph nodes.Nat.Rev.Immunol.3, 867–878.

34M.van den Heuvel et al./Journal of Reproductive Immunology67(2005)21–34

Warnock,R.A.,Campbell,J.J.,Dorf,M.E.,Matsuzawa,A.,McEvoy,L.M.,Butcher,E.C.,2000.The role of chemokines in the microenvironmental control of T versus B cell arrest in Peyer’s patch high endothelial venules.J.Exp.Med.191,77–88.

Watanabe,M.,Iwatani,Y.,Kaneda,T.,Hidaka,Y.,Mitsuda,N.,Morimoto,Y.,Amino,N.,1997.Changes in T, B,and NK lymphocyte subsets during and after normal pregnancy.Am.J.Reprod.Immunol.37,368–377. Yovel,G.,Shakhar,K.,Ben-Eliyahu,S.,2001.The effects of sex,menstrual cycle,and oral contraceptives on the number and activity of natural killer cells.Gynecol.Oncol.81,254–262.

丘吉尔演讲永不放弃

丘吉尔演讲永不放弃篇一：丘吉尔英语演讲稿：就职演说(中英对照) 1940年5月8日，由于前首相张伯伦遭到不信任质疑动议，被迫辞职。5月10日下午6时，国王召见丘吉尔，令其组阁；一小时后丘吉尔会见工党领袖艾德礼，邀请工党加入内阁并获得支持。3天后丘吉尔首次以首相身份出席下议院会议，发表了著名的讲话：“我没有别的，只有热血、辛劳、眼泪和汗水献给大家……你们问：我们的目的是什么？我可以用一个词来答复：胜利，不惜一切代价去争取胜利，无论多么恐怖也要争取胜利，无论道路多么遥远艰难，也要争取胜利，因为没有胜利就无法生存。”下议院最终以381票对0票的绝对优势表明了对丘吉尔政府的支持。正文： On Friday evening last I received from His Majesty the mission to form a new administration. 上星期五晚上，我奉陛下之命，组织新的一届政府。 It was the evident will of Parliament and the nation that this should be conceived on the broadest possible basis and that it should include all parties. 按国会和国民的意愿，新政府显然应该考虑建立在尽可能广泛的基础上，应该兼容所有的党派。 I have already completed the most important part of this task. A war cabinet has been formed of five members, representing, with the Labor, Opposition and Liberals, the unity of the nation. 我已经完成了这项任务的最主要的部分。战时内阁已由五人组成，包括工党、反对党和自由党，这体现了举国团结一致。 It was necessary that this should be done in one single day on account of the extreme urgency and rigor of events. Other key positions were filled yesterday. I am submitting a further list to the King tonight. I hope to complete the appointment of principal Ministers during tomorrow. 由于事态的极端紧急和严峻，新阁政府须于一天之内组成，其他的关键岗位也于昨日安排就绪。今晚还要向国王呈报一份名单。我希望明天就能完成几位主要大臣的任命。 The appointment of other Ministers usually takes a little longer. I trust when Parliament meets again this part of my task will be completed and that the administration will be complete in all respects. 其余大臣们的任命照例得晚一些。我相信，在国会下一次召开时，任命将告完成，臻于完善。

魔鬼恋人攻略全

魔鬼恋人攻略全文档编制序号：[KKIDT-LLE0828-LLETD298-POI08]

这是我一步步选出来的攻略，不过我只玩了最好的那个结局，所以选项都是选的加好感的哦，我没有玩坏的结局哦，希望大家多多支持我的攻略每个角色共有三个结局。开启条件分别为M度20/40/60（）。 Heaven剧本的解锁与Lock章节最后出现的关键词选择有关。注意： 1、长子、二子、末子，一周目无法攻略； 2、以下攻略lock部分选项均为全M选择肢，要开启其他路线重新读取章节选择另一选项即可； 3、肢节项改变后数值即更改，无需看完全部重复剧情。可直接选择Start-Chapter进行下一选项。逆卷绫人 Dark章 01因为很痛 02把毒吸出来太危险了 03总之先找找看吧 04我才不害怕呢 05谢谢你 06那我就用了哦 07照他说的做（单纯的进食，鲜血的味道） 08对不起…（第一次，他的快乐） 09把杯子递给他（麻痹，这种感觉） 10藏起玫瑰连珠（嫉妒，独占）

Maniac章 01蹲下 02把橡皮递给他 03四肢着地 04试试看吧 05腿 06让他尝尝味道 07那个…（绫人君的话，渐渐堕落）08放松身体（微笑，侵蚀） 09闭上眼睛（刻下，放弃） 10逃跑（吸血冲动，忍耐） Ecstasy章 01牵着手吧 02只是送他过去而已 03对不起 04说服他 05踢绫人一脚 06因为被吸血也不会死 07没事（名字，心脏）

08警戒绫人（魔王的血，她） 09结束广播（甜蜜的关系，捕食者） 10看着绫人（执着，这种关系）逆卷奏人 Dark章 01法式吐司 02对不起 03白棋子啊 04向声音传来的方向前进 05奏人君！ 06听理由 07不去（无法理解，不明白） 08没有那样的事（疼痛，快乐） 09谢谢你（蔷薇的香气，人的心情） 10对不起（心脏，散歩） Maniac章 01抚摸头部 02接近 03奏人君的东西 04散歩 05话题的故乡 06担心？ 07拥抱回报（不来看，手指的痕迹） 08不懂（不被原谅的行为，执着心） 09忘了（喜欢，在你身边） 10不错（那个人，取而代之的是） Ecstasy章 01食品 02沉默 03接吻 04什么演讲的？ 05再一次说 06为了奏人君 07可怕（喜欢，沉沦下去） 08伸出舌头（巧合造成的，永远） 09已经不说话（药，永远） 10寂寞（认真，相信）逆卷礼人 Dark章 01讨厌啊 02的完全不知道 03裸体围裙 04变态！！ 05饵食？ 06假装支付 07吸…（暗夜的居民，你） 08只好读（人类与恶魔的交易，逆卷到家里来了）09那个那样（监视谁，我的心脏）

丘吉尔演讲稿永不放弃

第一篇丘吉尔演讲稿永不放弃:丘吉尔演讲的启示决不能放弃温斯顿·丘吉尔，英国前首相，领导英国取得了第二次世界大战的胜利，是二十世纪中全球最重要的政治领袖之一。2002年，被英国BBC评选为“有史以来最伟大的英国人”。丘吉尔不仅是一位杰出的政治家，也是一位出色的演说家。面对山呼海啸一般的掌声，丘吉尔的演说，就是掠过风雨的惊雷、穿透黑夜的闪电，思想深邃，气势非凡，集聚了英国民众的智慧与力量，道德与情感。丘吉尔的政治生涯中，作过无数次精彩绝伦的演讲。今天的人们，用平常的笔墨哪里能传神地描述当年演讲者的气质与风采，以及演讲场面的壮阔与轰动。在战争阴云密布，面对法西斯的毁灭性进攻时，丘吉尔向英国也是向全世界作了二战时期极为著名的战争动员演讲“我们将战斗到底。我们将在法国作战，我们将在海洋中作战，我们将以越来越大的信心和越来越强的力量在空中作战，我们将不惜一切代价保卫本土，我们将在海滩作战，我们将在敌人的登陆点作战，我们将在田野和街头作战，我们将在山区作战。我们绝不投降！”钢铁般的声音，坚定地展示了一个领袖人物的钢铁意志。作为一个战争英雄，虽然在战后失去了首相的职务。然而面对政治的落寞和失意，丘吉尔极有尊严地用古希腊作家普鲁塔克的一句名言告别了政坛“对他们的伟大人物忘恩负义，是伟大民族的标志”，简短的语言，让人肃然起敬，这样的思想与情怀，一般的人又如何能理解？丘吉尔在牛津大学演讲，则又是另外一种风采。这种风采不仅震撼了牛津，也震撼了多少不同国籍的后人。1948年，他应邀在该大学作一个主题为“成功秘诀”的专题讲座，面对充满期望的牛津学子和全世界各大新闻媒体，丘吉尔作了一个极为简短却寓意深刻的演讲，不妨全文摘录“我的成功秘诀有三个第一是，决不放弃；第二是，决不、决不放弃；第三是，决不、决不、决不能放弃！我的演讲结束了。”会场一片凝重，丘吉尔以最简洁的语言、最执着的意志，揭示了最深刻的人生哲理。听众在沉寂了足足一分钟后，对着已经没有演讲者的讲台，爆发出如雷般的掌声。从演讲者来说，这就是最高的奖赏。人若能畅快淋漓地表达自己的思想与感情，且你的思想与感情，得到成千上万人的呼应与融合，这该是何等的痛快与荣耀。丘吉尔的一生，可谓波澜壮阔，在两次世界大战中叱咤风云，出生入死，是战争中的传奇英雄，又是享有世界级声誉的政治家，还是深受读者欢迎的作家、画家，获得过诺贝尔文学奖。如此出类拔萃，自然赢得了英国人民的爱戴，国家为他而骄傲。牛津大学请他作成功秘诀的演讲，当然非他莫属。丘吉尔何以成功？他无论从哪个方面都可以洋洋洒洒地说上半天，然而他却出人意料地只是以三句话、为时三分钟的闪电式演讲，概括了自己艰难而又卓绝的人生要义，留给听众闪电般的力量。闪电，只是一瞬间，却光耀长空，精彩的演讲，也是如此，最短促的，往往却是最长久的，而最长久的，却一定是最不朽的。丘吉尔辉煌一生，尽可以导师的身份，向青年学子展示自己的人生经历，然而他避而不谈自己的成功，把“成功秘诀”也仅仅谦虚地归结为“我的”，既然是“我的”，于大众而言，只是成功之路的参考。其实，最自我的，往往是最人性的，而最人性的，却一定是最人类的。这大概也是丘吉尔演讲堪称经典的“秘诀”所在。丘吉尔著作等身，文笔一流，掌握近五万多词汇，被传为历史上掌握英语单词词汇量最多的人之一。然而，这篇演讲的核心，他只用了三个英语单词“决不放弃(Never give up)”。最简洁，往往最隽永，而最隽永的，却一定是最丰富的。倘若每一个人都能选定适合自己的目标而“决不放弃”，人生又该是何等的精彩。激情的时代，一定有激情的演讲。而激情的演讲，一定是奔放自己的思想、意志与情感的结晶。激情的演讲，是时代的标记，是历史的财富。

铁血泪——丘吉尔演讲词中英对照

铁血泪——丘吉尔演讲词中英对照.txt10有了执著，生命旅程上的寂寞可以铺成一片蓝天；有了执著，孤单可以演绎成一排鸿雁；有了执著，欢乐可以绽放成满圆的鲜花。铁血泪BLOOD, SWEAT AND TEARS" (温斯顿丘吉尔的演讲) Winston Churchill (May 13, 1940 ) On Friday evening last I received from His Majesty the mission to form a new administration. It was the evident will of Parliament and the nation that this should be conceived on the broadest possible basis and that it should include all parties. I have already completed the most important part of this task. A war cabinet has been formed of five members, representing, with the Labor, Opposition and Liberals, the unity of the nation. It was necessary that this should be done in one single day on account of the extreme urgency and rigor of events. Other key positions were filled yesterday. I am submitting a further list to the King tonight. I hope to complete the appointment of principal Ministers during tomorrow. The appointment of other Ministers usually takes a little longer. I trust when Parliament meets again this part of my task will be completed and that the administration will be complete in all respects. I considered it in the public interest to suggest to the Speaker that the House should be summoned today. At the end of today's proceedings, the adjournment of the House will be proposed until May 2l with provision for earlier meeting if need be. Business for that will be notified to M. P. 's at the earliest opportunity. I now invite the House by a resolution to record its approval of the steps taken and declare its confidence in the new government. The resolution: "That this House welcomes the formation of a government representing the united and inflexible resolve of the nation to prosecute the war with Germany to a victorious conclusion." To form an administration of this scale and complexity is a serious undertaking in itself. But we are in the preliminary Phase of one of the greatest battles in history. We are in action at any other points-in Norway and in Holland-and we have to be prepared in the Mediterranean. The air battle is continuing, and many preparations have to be made here at home. In this crisis I think I may be pardoned if I do not address the House at any length today, and I hope that any of my friends and colleagues or for mer colleagues who are affected by the political reconstruction will make all allowances for any lack of ceremony with which it has been necessary to act. I say to the House as I said to Ministers who have joined this government, I have nothing to offer but blood, toil, tears and sweat. We have before us an ordeal of the most grievous kind. We have before us many, many months of struggle and suffering. You ask, what is our policy? I say it is to wage war by land, sea and air. War with all our might and with all the strength God has given us, and to wage war against a monstrous tyranny never surpassed in the dark and lamentable catalogue of human crime. That is our policy. You ask, what is our aim? I can answer in one word, It is victory. Victory at all

魔鬼恋人攻略(全)

这是我一步步选出来的攻略，不过我只玩了最好的那个结局，所以选项都是选的加好感的哦，我没有玩坏的结局哦，希望大家多多支持我的攻略每个角色共有三个结局。开启条件分别为M度20/40/60（？）。 Heaven剧本的解锁与Lock章节最后出现的关键词选择有关。注意： 1、长子、二子、末子，一周目无法攻略； 2、以下攻略lock部分选项均为全M选择肢，要开启其他路线重新读取章节选择另一选项即可； 3、肢节项改变后数值即更改，无需看完全部重复剧情。可直接选择Start-Chapter进行下一选项。逆卷绫人 Dark章 01因为很痛 02把毒吸出来太危险了 03总之先找找看吧 04我才不害怕呢 05谢谢你 06那我就用了哦 07照他说的做（单纯的进食，鲜血的味道） 08对不起…（第一次，他的快乐） 09把杯子递给他（麻痹，这种感觉） 10藏起玫瑰连珠（嫉妒，独占） Maniac章 01蹲下 02把橡皮递给他 03四肢着地 04试试看吧 05腿 06让他尝尝味道 07那个…（绫人君的话，渐渐堕落） 08放松身体（微笑，侵蚀） 09闭上眼睛（刻下，放弃） 10逃跑（吸血冲动，忍耐） Ecstasy章 01牵著手吧 02只是送他过去而已 03对不起 04说服他 05踢绫人一脚 06因为被吸血也不会死 07没事（名字，心脏） 08警戒绫人（魔王的血，她） 09结束广播（甜蜜的关系，捕食者） 10看着绫人（执着，这种关系）逆卷奏人 Dark章 01法式吐司 02对不起 03白棋子啊 04向声音传来的方向前进 05奏人君！ 06听理由 07不去（无法理解，不明白） 08没有那样的事（疼痛，快乐） 09谢谢你（蔷薇的香气，人的心情） 10对不起（心脏，散歩） Maniac章 01抚摸头部 02接近 03奏人君的东西 04散歩 05话题的故乡 06担心？ 07拥抱回报（不来看，手指的痕迹）

丘吉尔演讲稿

丘吉尔演讲稿《Blood,Toil.Tears and Sweet》，《We shall fight them on the beachers》，《Iron Curtain.Speach(铁幕演说）》，三篇演讲稿英文及其翻译。这是丘吉尔1940年5月13日在下院发表的首相就职演讲，内容大概如下： “我所能奉贤的，只有热血和辛劳，眼泪和汗水。你们问：我们的政策是什么？我说，我们的政策就是用上帝所给予我们的全部能力和全部力量，在海上.陆地上和空中进行战争。同一个邪恶悲惨的人类罪恶史上从为见过的穷凶极恶的暴政进行战争。你们问：我们的目的是什么？我可以用一个词来答复：胜利————不惜一切代价去争取胜利，无论多么恐怖也要去争取胜利；无论道路多么遥远和艰难，也要去争取胜利；因为没有胜利，就不能生存。在这个时候，我觉得我有权要求大家的支持，我说：起来，让我们联合起来，共同前进！” 热血、辛劳、眼泪和汗水（1940.5.13）丘吉尔星期五晚上，我接受了英王陛下的委托，组织新政府。这次组阁，应包括所有的政党，既有支持上届政府的政党，也有上届政府的反对党，显而易见，这是议会和国家的希望与意愿。我已完成了此项任务中最重要的部分。战时内阁业已成立，由5位阁员组成，其中包括反对党的自由主义者，代表了举国一致的团结。三党领袖已经同意加入战时内阁，或者担任国家高级行政职务。三军指挥机构已加以充实。由于事态发展的极端紧迫感和严重性，仅仅用一天时间完成此项任务，是完全必要的。其他许多重要职位已在昨天任命。我将在今天晚上向英王陛下呈递补充名单，并希望于明日一天完成对政府主要大臣的任命。其他一些大臣的任命，虽然通常需要更多一点的时间，但是，我相信会议再次开会时，我的这项任务将告完成，而且本届政府在各方面都将是完整无缺的。我认为，向下院建议在今天开会是符合公众利益的。议长先生同意这个建议，并根据下院决议所授予他的权力，采取了必要的步骤。今天议程结束时，建议下院休会到5月21日星期二。当然，还要附加规定，如果需要的话，可以提前复会。下周会议所要考虑的议题，将尽早通知全体议员。现在，我请求下院，根据以我的名义提出的决议案，批推已采取的各项步骤，将它记录在案，并宣布对新政府的信任。组成一届具有这种规模和复杂性的政府，本身就是一项严肃的任务。但是大家一定要记住，我们正处在历史上一次最伟大的战争的初期阶段，我们正在挪威和荷兰的许多地方进行战斗，我们必须在地中海地区做好准备，空战仍在继续，众多的战备工作必须在国内完成。在这危急存亡之际，如果我今天没有向下院做长篇演说，我希望能够得到你们的宽恕。我还希望，因为这次政府改组而受到影响的任何朋友和同事，或者以前的同事，会对礼节上的不周之处予以充分谅解，这种礼节上的欠缺，到目前为止是在所难免的。正如我曾对参加本届政府的成员所说的那样，我要向下院说：“我没什么可以奉献，有的．只是热血、辛劳、眼泪和汗水。” 摆在我们面前的，是一场极为痛苦的严峻的考验。在我们面前，有许多许多漫长的斗争和苦难的岁月。你们问：我们的政策是什么我要说，我们的政策就是用我们全部能力，用上帝所给予我们的全部力量，在海上、陆地和空中进行战争，同一个在人类黑暗悲惨的罪恶史上所

丘吉尔最短演讲

丘吉尔最短演讲丘吉尔拄着拐杖，戴着草帽，慢步走向讲台，先把草帽放在讲台，然后从左到右横扫了整个军营，说：“永不放弃！”然后又从左到右横扫了整个军营，说：“永不放弃！”当时整个军营鸦雀无声，连一根针掉在地上的声音都可以听到。然后他又从左到右横扫了一次整个军营，加大声量说：“永不放弃，永不放弃，永不放弃，永不放弃！”整个军营都兴奋起来，欢呼声和拥抱淹没了整个军营。此后英国连连打败了德国希特勒的进攻。这就是丘吉尔最著名的演讲，世界上最震撼的演讲，同时也是世界上最短的演讲。上星期五晚上，我接受了英王陛下的委托，组织新政府。这次组阁，应包括所有政党，既有支持上届政府的政党，也有上届政府的反对党，显而易见，这是议会和国家的希望与意愿。我已完成了此项任务中最重要的部分。战时内阁业已成立，由5位阁员组成，其中包括反对党的自由主义者，代表举国一致的团结。三党领袖已经同意加入战时内阁，或者担任国家高级行政职务。三军指挥机构已加以充实。由于事态发展的极端紧迫感和严重性，仅仅用一天时间完成此项任务，是完全必要的。其他许多重要职位在昨天任命。我将在今天晚上向英王陛下呈递补充名单，并希望于明日一天完成对政府主要大臣的任命。其他一些大臣的任命，虽然通常需

要更多一点的时间，但是，我相信会议再次开会时，我的这项任务将告完成，而且本届政府在各方面都将是完整无缺的。我认为，向下院建议在今天开会是符合公众利益。议长先生同意这个建议，并根据下院决议所授予他的权力，采取了必要的步骤。今天议程结束时，建议下院休会到5月21日星期二。当然，还要附加规定，如果需要的话，可以提前复会。下周会议所要考虑的议题，将尽早通知全体议员。现在，我请求下院，根据以我的名义提出的决议案，批准已采取的各项步骤，将它记录在案，并宣布对新政府的信任。组成一届具有这种规模和复杂性的政府，本身就是项严肃的任务。但是大家一定要记住，我们正处在历史上一次最伟大的战争的初期阶段，我们正在挪威和荷兰的许多地方进行战斗，我们必须在地中海地区做好准备，空战仍在继续，众多的战备工作必须在国内完成。在这危急存亡之际，如果我今天没有向下院做长篇演说，我希望能够得到你们的宽耍我还希望，因为这次政府改组而受到影响的任何朋友和同事，或者以前的同事，会对礼节上的不周之处予以充分谅解，这种礼节上的欠缺，到目前为止是在所难免的。正如我曾对参加本届政府的成员说的那样，我要向下院说：“我没有什么可以奉献，有的只是热血、辛劳、眼泪和汗水。” 摆在我们面前的，是一场极为痛苦的严峻的考验。在我

丘吉尔演讲语录

丘吉尔演讲语录导读：丘吉尔英国政治家、历史学家、那么你对丘吉尔了解吗？下面一起去阅读小编为你整理的丘吉尔演讲语录吧！丘吉尔演讲语录1、决定不做决定，决心动摇不定，坚定地随波逐流，稳稳地随风摇曳，强大到一无所成。 2、如果我们让过去和现在争吵，我们将失去未来。 3、成功就是不断失败不失信心。 4、为了得到真正的快乐，避免烦恼和脑力的过度紧张，我们都应该有一些嗜好。 5、对他们的伟大人物忘恩负义，是伟大民族的标志。 6、永不，永不，永不屈服。 7、简短的词是最好的，又短又古老的词最最好。 8、开玩笑是件严肃的事情。 9、口才艺术主要分四大要素，而语速占第一位。 10、成功不要紧，失败不致命。继续前行的勇气，才最可贵。 11、一个人绝对不可在遇到危险的威胁时，背过身去试图逃避。若是这样做，只会使危险加倍。但是如果立即面对它毫不退缩，危险便会减半。 12、当老鹰沉默时，鹦鹉就开始叽叽喳喳了。 13、悲观主义者在每个机会里看到困难。乐观主义者在每个困难里看到机会。

14、谎言环游全球的时候，真相还没穿好裤子。 15、美德与伟大不能兼得。 16、我本来愿意显露一下自己的学识的，而他们则千方百计地揭露我的无知。 17、如果你指定一万条规章，你就摧毁了所有对法律的尊重。 18、现在不是轻松舒适的时候。现在是大胆和忍耐的时候。 19、反对民煮最好的论据就是与一个普通选民五分钟的谈话。 20、资本主义的邪恶在于不公平的幸福分配。涉汇煮意的美德在于公平享受痛苦。 21、钱就像肥料，广施才有效。 22、妥协的人是在喂一条鳄鱼希望它最后吃掉自己。 23、当我们跪下去的时候，伟大领袖便产生了。当我们不会反抗的时候，奴隶便产生了。当我们不会质疑，骗子便产生了。当我们太娇惯，畜生便产生了。 24、我们靠得到的东西生存，但是靠给予的东西生活。 25、最可怕的事情就是不容忍的乌托邦。 26、民煮是最不坏的制度。 27、破坏容易建设难。 28、我"吃"文字从来没反胃过。 29、不管任何代价，我们一定要生存下去。 30、人经常被真相困扰。但是大多数人装作没看见就走了。

丘吉尔演讲稿

丘吉尔演讲稿《Blood, and Sweet》，《We shall fight them on the beachers》，《Iron (铁幕演说）》，三篇演讲稿英文及其翻译。这是丘吉尔1940年5月13日在下院发表的首相就职演讲，内容大概如下： “我所能奉贤的，只有热血和辛劳，眼泪和汗水。你们问：我们的政策是什么我说，我们的政策就是用上帝所给予我们的全部能力和全部力量，在海上.陆地上和空中进行战争。同一个邪恶悲惨的人类罪恶史上从为见过的穷凶极恶的暴政进行战争。你们问：我们的目的是什么我可以用一个词来答复：胜利————不惜一切代价去争取胜利，无论多么恐怖也要去争取胜利；无论道路多么遥远和艰难，也要去争取胜利；因为没有胜利，就不能生存。在这个时候，我觉得我有权要求大家的支持，我说：起来，让我们联合起来，共同前进！” 热血、辛劳、眼泪和汗水（）丘吉尔星期五晚上，我接受了英王陛下的委托，组织新政府。这次组阁，应包括所有的政党，既有支持上届政府的政党，也有上届政府的反对党，显而易见，这是议会和国家的希望与意愿。我已完成了此项任务中最重要的部分。战时内阁业已成立，由5位阁员组成，其中包括反对党的自由主义者，代表了举国一致的团结。三党领袖已经同意加入战时内阁，或者担任国家高级行政职务。三军指挥机构已加以充实。由于事态发展的极端紧迫感和严重性，仅仅用一天时间完成此项任务，是完全必要的。其他许多重要职位已在昨天任命。我将在今天晚上向英王陛下呈递补充名单，并希望于明日一天完成对政府主要大臣的任命。其他一些大臣的任命，虽然通常需要更多一点的时间，但是，我相信会议再次开会时，我的这项任务将告完成，而且本届政府在各方面都将是完整无缺的。我认为，向下院建议在今天开会是符合公众利益的。议长先生同意这个建议，并根据下院决议所授予他的权力，采取了必要的步骤。今天议程结束时，建议下院休会到5月21日星期二。当然，还要附加规定，如果需要的话，可以提前复会。下周会议所要考虑的议题，将尽早通知全体议员。现在，我请求下院，根据以我的名义提出的决议案，批推已采取的各项步骤，将它记录在案，并宣布对新政府的信任。组成一届具有这种规模和复杂性的政府，本身就是一项严肃的任务。但是大家一定要记住，我们正处在历史上一次最伟大的战争的初期阶段，我们正在挪威和荷兰的许多地方进行战斗，我们必须在地中海地区做好准备，空战仍在继续，众多的战备工作必须在国内完成。在这危急存亡之际，如果我今天没有向下院做长篇演说，我希望能够得到你们的宽恕。我还希望，因为这次政府改组而受到影响的任何朋友和同事，或者以前的同事，会对礼节上的不周之处予以充分谅解，这种礼节上的欠缺，到目前为止是在所难免的。正如我曾对参加本届政府的成员所说的那样，我要向下院说：“我没什么可以奉献，有的．只是热血、辛劳、眼泪和汗水。” 摆在我们面前的，是一场极为痛苦的严峻的考验。在我们面前，有许多许多漫长的斗争和苦难的岁月。你们问：我们的政策是什么我要说，我们的政策就是用我们全部能力，用上帝所给予我们的全部力量，在海上、陆地和空中进行战争，同一个在人类黑暗悲惨的罪恶史上所

丘吉尔在美国度圣诞节的即兴演讲汇编

---------------------------------------------------------------范文最新推荐------------------------------------------------------ 丘吉尔在美国度圣诞节的即兴演讲各位为自由而奋斗的劳动者和将士：我的朋友，伟大而卓越的罗斯福总统，刚才已经发表过圣诞前夕的演说，已经向全美国的家庭致友爱的献词。我现在能追随骥尾讲几句话，内心感到无限的荣幸。我今天虽然远离家庭和祖国，在这里过节，但我一点也没有异乡的感觉。我不知道，这是由于本人母亲的血统和你们相同，抑或是由于本人多年来在此地所得的友谊，抑或是由于这两个文字相同、信仰相同、理想相同的国家，在共同奋斗中所产生出来的同志感情，抑或是由于上述三种关系的综合。总之，我在美国的政治中心地——华盛顿过节，完全不感到自己是一个异乡之客。我和各位之间，本来就有手足之情，再加上各位欢迎的盛意，我觉得很应该和各位共坐炉边，同享这圣诞之乐。但今年的圣诞前夕，却是一个奇异的圣诞前夕。因为整个世界都卷入了一种生死搏斗之中，使用着科学所能设计的恐怖武器来互相屠杀。假若我们不是深信自己对别国领土和财富没有贪图的恶意，没有攫取物资的野心，没有卑鄙的念头，那么我们今年的圣诞节，一定很难过。 1 / 16

战争的狂潮虽然在各地奔腾，使我们心惊肉跳，但在今天，每一个家庭都在宁静的、肃穆的气氛里过节。今天晚上，我们可以暂且把恐惧和忧虑抛开、忘记，而为那些可怜的孩子们布置一个快乐的晚会。全世界说英语的家庭，今晚都应该变成光明的和平的小天地，使孩子们尽量享受这个良宵，使他们因为得到父母的恩物而高兴，同时使我们自己也能享受这种无牵无挂的乐趣，然后我们担起明年艰苦的任务，以各种的代价，使我们孩子所应继承的产业，不致被人剥夺；使他们在文明世界中所应有的自由生活，不致被人破坏。因此，在上帝庇佑之下，我谨祝各位圣诞快乐。市上半年经济形势调查与思考市上半年经济形势调查与思考今年以来，全市各级各部门认真贯彻市委、市政府“工业强市、民营富民”的发展战略，按照市四届人大三次会议确定的工作目标和任务，牢牢把握全面发展靠经济、经济发展靠工业、工业发展靠民营靠招商、民营和招商靠环境这条主线，开拓进取，扎实工作，保持了经济较快增长、社会稳定进步的良好势头。一、上半年经济运行主要特点上半年，我市经济在较高平台上保持了持续稳定增长的态势，全市实现地区生产总值275.3亿元，同比增长10.8。其中，第一产业增加值50.02亿元，第二产业增加值123.46亿元，第三产业增加值101.81亿元，分别增长5、12.9、10.9。——从生产领域看：农业总量增加、结构变化。上半年全

魔鬼恋人攻略(全

欢迎阅读这是我一步步选出来的攻略，不过我只玩了最好的那个结局，所以选项都是选的加好感的哦，我没有玩坏的结局哦，希望大家多多支持我的攻略每个角色共有三个结局。开启条件分别为M度20/40/60（？）。 Heaven剧本的解锁与Lock章节最后出现的关键词选择有关。注意： 1、长子、二子、末子，一周目无法攻略； 2、以下攻略lock部分选项均为全M选择肢，要开启其他路线重新读取章节选择另一选项即可； 3、肢节项改变后数值即更改，无需看完全部重复剧情。可直接选择Start-Chapter进行下一选项。逆卷绫人 Dark章 01 02 03 04 05谢谢你 06 07 08 09 10 Maniac章 01蹲下 02把橡皮递给他 03四肢着地 04试试看吧 05腿 06让他尝尝味道 07那个…（绫人君的话，渐渐堕落） 08放松身体（微笑，侵蚀）

09闭上眼睛（刻下，放弃） 10逃跑（吸血冲动，忍耐） Ecstasy章 01牵着手吧 02只是送他过去而已 03对不起 04说服他 06担心？ 07拥抱回报（不来看，手指的痕迹）08不懂（不被原谅的行为，执着心）09忘了（喜欢，在你身边） 10不错（那个人，取而代之的是） Ecstasy章 01食品 02沉默 03接吻 04什么演讲的？ 05再一次说 06为了奏人君 07可怕（喜欢，沉沦下去）

08伸出舌头（巧合造成的，永远） 09已经不说话（药，永远） 10寂寞（认真，相信）逆卷礼人 Dark章 01讨厌啊 02的完全不知道 03裸体围裙 04变态！！ 05饵食？ 06假装支付 07吸…（暗夜的居民，你） 08只好读（人类与恶魔的交易，逆卷到家里来了）09那个那样（监视谁，我的心脏） 10没办法（饲料有玩具，关系很差的） Maniac章 01讨厌！ 02舒服 03 04回头 05明白了 06 07 08 09 10 Ecstasy章 01不服吗 02勉强了。 03抵抗 04 05高兴 06放弃 07 08 10 逆卷昂 Dark章 01凝视 02抵抗 03可爱 04 05道谢 06 07 08 09，从背后…（吐息混杂的私语，背德感） 10自己动（独占欲，笨拙） Maniac章 01无聊吗？ 02不选 03低头道歉 04咬住 05老实说 06逃跑 07那样的讨厌（无条件，过剩） 08起来（隐藏的，开始感到了的心） 09斯巴鲁君身（热，本质） 10扭头（说，悲哀）

2020年永不言弃演讲稿三分钟模板

永不言弃演讲稿三分钟模板能否经受得起考验，不单单看你的本领，还要靠你的毅力，毅力是决定成败的关键。成功的事实告诉我们永不言弃是一种品格，更是一种精神，只有永不言弃的人才有胜利的希望。一起来看看永不言弃演讲稿三分钟，欢迎查阅！永不言弃演讲稿三分钟1 尊敬的老师们，亲爱的同学们大家好！我们每一个人在成长的时候，都会遇到大大小小的挫折，这对于我们来说，是属于正常的，毕竟成长路上挫折多。我记得初一刚进校的时候，我便遇到了很大的问题。我的数学一直在6几分这里徘徊，我从来都未曾到过9几分。但过了一段时间，我开始变得很努力，在家里努力地做着题目，做着做着，我便明白了其中的道理，遇到困难并不可怕，但是我们面对挫折要坚定、执着，不能因为遇到它就开始害怕了，这样子的话，长大以后便会一事无成，遇到困难就退缩。努力终于没有白费，我凭借做题目，居然在考试中遇到了相同的题目，我便轻而易举地做起来了。毕竟我是经历过挫折的人，成长的路上是不可能没有挫折，但正因为有挫折才会使我们变得强大起来。还有一次，我和父母去野炊，我负责将火给点起来。于是，我想了钻木取火的方法。我的力气可能还不够大，但是，只要我用尽，就一定可以成功的。于是，我先拿了一根很粗的树枝，钻呀，钻呀，过了一段时间，便有了烟，可是还没有火。我继续努力地钻着，渐渐地有了火苗，成功了！大火在树桩烧了起来，这下子，终于可以做饭了。所以说，我们在成长的途中怎么可能不会遇到挫折，但是要正确地面对它，用自己的大脑去解决，人生的路上挫折多，正因为挫折，使得我们有了提升的空间，让我们变得更加强大。我一直都坚信着这个道理，成长路上挫折多。永不言弃演讲稿三分钟2 尊敬的老师们，亲爱的同学们大家好！秋。满树的黄花如一颗颗黄宝石缀满了枝头，阵阵微风拂过，把那些摇摇欲坠的桂花夹带在风中，最终撒落在如一层金地毯的大地上，眼前顿时一片金黄。可这样的美景，也解不开我心中的解。

坚持到底,永不放弃演讲

演讲英国首相丘吉尔有一次著名的演讲，那是她生命中的左后一次演讲，在一所大学的毕业典礼上，演讲总共20分钟，内容只有两句话，坚持到底，永不放弃，坚持到底，永不放弃，短短的两句话，包含着首相的苦心和希望，回荡在每一个人的心里，那是一个能够载入试车的讲演，也是丘吉尔一生心血的凝聚，是他一生的经验，成功需要精准的眼光，强大的行动力还有毅力，用果决的行动抓住机遇，用坚持和毅力换取成功，有人这样比喻着，人生有两杯水，一杯甘甜，一杯苦涩，你是什么人，就看你如何喝掉这两杯水，一种人先喝掉苦水，再喝掉甜水，后来，他们成为成功者，另一种人，先喝掉甜水，再喝掉苦水，后来他们庸碌无为，面对挫折，让自己再坚持一下，失败就过去了，所谓失败是成功之母，只有经历失败才会走向成功，成功者与失败者的区别仅仅在于站起来不倒下多一次或少一次，仅仅一次的差别，最后一次，你能否爬起来？丘吉尔，乱世中的英雄，二战期间领到英国胜利的首相，谈到成功，总结其毕生的经验，也不过两句话，坚持到底，永不放弃，有人说，敏锐的目光，果断地行动是成功的条件，还有重要的是坚韧的毅力，一帆风顺的人并不存有，面对挫折，我们最坚实的后盾就是毅力，拿破仑说，我只有一个忠告给你——做你自己的主人。没有高贵的出身，没有绝色的容貌，没有特别的天赋，有成功，便只有靠自己，坚韧的毅力是我们唯一且坚实的后盾，丘吉尔以前也是大家眼里的问题少年，多

少年成绩不佳，甚至还经常遭到老师的体罚，他没有被打垮，反而奋起，终于，在持续拼搏中的丘吉尔步入政坛，直到坐上首相这把交椅。人生小哲理，每个人都是自己的主人，毅力是我们坚强的后盾，世界上没有失败的人，只有尚未成功的人，成功无他，坚持而已，很多时候，我们被迫放弃在成功面前，却在事实面前追悔莫及，仅仅最后一点点的坚持，决定了一个人的成败，莫要被自己的心里吓退，莫要档成功近在咫尺时倒下，在努力一点，再坚持一下，梦想会变成现实。

丘吉尔演讲永不放弃(WORD可编辑).docx

【演讲稿模板】丘吉尔演讲永不放弃编写人：XXX 日期：XXX Word A4打印标准格式可随意修改

I have already completed the most important part of this task. A war cabinet has been formed of five members, representing, with the Labor, Opposition and Liberals, the unity of the nation. 我已经完成了这项任务的最主要的部分。战时内阁已由五人组成，包括工党、反对党和自由党，这体现了举国团结一致。 It was necessary that this should be done in one single day on account of the extreme urgency and rigor of events. Other key positions were filled yesterday. I am submitting a further list to the King tonight. I hope to complete the appointment of principal Ministers during tomorrow. 由于事态的极端紧急和严峻，新阁政府须于一天之内组成，其他的关键岗位也于昨日安排就绪。今晚还要向国王呈报一份名单。我希望明天就能完成几位主要大臣的任命。 The appointment of other Ministers usually takes a little longer. I trust when Parliament meets again this part of my task will be completed and that the administration will be complete in all respects. 其余大臣们的任命照例得晚一些。我相信，在国会下一次召开时，任命将告完成，臻于完善。

魔鬼爱人游戏中文攻略

魔鬼爱人psp游戏中文攻略：[1]长子逆卷修 ? ?| ?浏览：6431 ?| ?更新：2013-10-17 20:56 ?1 ?2 ?3 ?4 ?5 ?6 ?7 分步阅读

看了近十年动漫的外貌协会重口味……最近果然又控制不住自己，跑去下了psp 模拟器就为了玩这个魔鬼爱人。玩游戏果然还是需要攻略的啊……特别是类似于这种文字恋爱养成，厨娘觉得更多的是为了剧情才玩这个游戏吧……长子逆卷修，先来一炮。方法/步骤 1.psp模拟器键位使用：（没有用过psp模拟器的基友们可以看一看这个步骤，老鸟直接跳过吧。）前面的解说什么的可以用【空格键】跳掉。Z：是退出或者返回。X：是确认。这个三个键必须知道，不然你会觉得无比疼痛。方向键控制箭头。A：切换画面速度。W：切换章节（魔鬼恋人分为DARK、Maniac、Ecstasy三个章节）。 2.玩之前大家要看一下这个，不然可能要重新玩。这里有四个图，大家仔细看 wow。第一个图：吸爱度是指好感度，S：攻值、M：受值（很奇怪，M全满就是好结局，S全满就是悲剧，一半一半的时候就变成还不错的结局。）

第二个图：这个东西还没有碰到之前不知道丫是神马……一般在每个章节的第七个到第十个部分出现。有三个选项选两个对的（不要按照常理来选看攻略吧），选对了会有出血动画。第三和第四图：第三个图用方向键选择，然后看到第四个图那个蓝色的灯没有，这个表示你选的选项是+M值的，反之就会出现+S值的紫色灯。

3.魔鬼爱人的每个人物都有三个结局可以攻略：best end（俗称最好的结局）、 better end（过得去的结局）、sad end（悲剧）。