Immuno Res_2014_review on__ inflammatory diseases_Hirsch
IMMUNOLOGY AT THE UNIVERSITY OF IOWA
Follistatin-like protein 1and its role in in?ammation and in?ammatory diseases
Yury Chaly ?Bruce Hostager ?Sonja Smith ?
Raphael Hirsch
óSpringer Science+Business Media New York 2014
Abstract Follistatin-like protein 1(FSTL1)is a secreted glycoprotein produced mainly by cells of mesenchymal origin.FSTL1has been shown to play an important role during embryogenesis;FSTL1-de?cient mice die at birth from multiple developmental abnormalities.In the last decade,FSTL1has been identi?ed as a novel in?ammatory protein,enhancing synthesis of proin?ammatory cytokines and chemokines by immune cells in vitro and in vivo.FSTL1mediates proin-?ammatory events in animal models of in?ammatory diseases,particularly in collagen-induced arthritis in mice.FSTL1is elevated in various in?ammatory conditions and decreased during the course of treatment.FSTL1may therefore be a valuable biomarker for such diseases.Moreover,a variety of experiments suggest that targeting of FSTL1may be useful in the treatment of diseases in which in?ammation plays a central role.Keywords
Follistatin-like protein 1áCytokines áMonocytes áMacrophages áIn?ammasome
Introduction
Follistatin-like protein 1(FSTL1),a secreted glycoprotein,also named TSC-36[transforming growth factor (TGF)b 1-stimulated clone 36]or FRP (follistatin-related protein),was originally cloned from the mouse osteoblastic cell line MC3T3-E1[1].FSTL1is widely expressed in human and mouse tissues,although its expression was found to be restricted to cells of non-hematopoietic lineage,in partic-ular to cells of the mesenchymal lineage [2–6].Recent studies have demonstrated that
cells of the mesenchymal lineage may actively regulate immune cell function and exert both pro-and anti-in?ammatory effects [7–9].These effects are mediated by various soluble factors released by cells within the in?ammatory milieu.
Although the function of FSTL1is not completely understood,it appears to play roles in the regulation of cell
survival,proliferation,differentiation,migration,organ development,carcinogenesis,and metastasis [10–14].FSTL1knockout (KO)mice die at birth and display mul-tiple developmental abnormalities of the respiratory and skeletal system [2,3,15].It has been shown that FSTL1affects mesenchymal tissue development by regulating signaling induced by proteins of the TGF b superfamily [2,3,15].
In the past decade,a number of reports have been published,suggesting a role for FSTL1in in?ammation and the regulation of immune cell function.Speci?cally,several studies have demonstrated that FSTL1is important in the pathogenesis of rheumatoid arthritis (RA)and other autoimmune diseases.In this review,we summarize the literature on potential roles of FSTL1in the in?ammatory response.
FSTL1gene structure
FSTL1is encoded on chromosome 16in mice and on chromosome 3in humans.The human FSTL1gene con-sists of 11exons (exon 1is noncoding)spanning nearly
Y.Chaly (&)áB.Hostager áS.Smith áR.Hirsch
Stead Family Department of Pediatrics,University of Iowa Carver College of Medicine,2191ML,500Newton Road,Iowa City,IA 52242,USA
e-mail:yury-chaly@https://www.wendangku.net/doc/2e17856998.html,
Raphael Hirsch
Yury Chaly
Immunol Res
DOI 10.1007/s12026-014-8526-z
60kb [14].Sundaram et al.[14]demonstrated that in humans,the FSTL1transcript encodes two products,the FSTL1protein and microRNA-198(miR-198).The latter is encoded in exon 11of the human FSTL1gene.The miR-198was shown to inhibit migration of keratinocyte and wound repair,whereas FSTL1exerted the opposite effect [14].
FSTL1protein structure and domains
The FSTL1protein is
308amino acids in length with an N-terminal signal peptide of 20amino acids.The non-gly-cosylated protein has a predicted molecular weight of 35kDa.The protein is found in two differentially glycos-ylated isoforms with similar functional activity [16].Zwijsen et al.[17]and Tanaka et al.[4]documented the molecular weights of different isoforms of FSTL1ranging from 40to 55kDa.Three putative N-glycosylation sites and phosphorylation sites have been predicted [17].Zwijsen et al.[17]did not reveal any membrane-spanning or membrane-associated sequences in the FSTL1molecule.FSTL1is highly conserved across mam-malian species with the human and mouse FSTL1proteins sharing 91%homology in their amino acid sequence [18].Follistatin-like protein 1contains a follistatin-like domain and an extracellular calcium-binding domain (Fig.1).Based on protein domain similarity between FSTL1and secreted protein acidic and rich in cysteine (SPARC/BM-40/osteonectin),FSTL1belongs to the SPARC family of proteins.The family also includes BM-40/SPARC/osteonectin,follistatin-like protein 4,follista-tin-like protein 5,SC1/hevin,testican-1,testican-2,testi-can-3,SMOC1,and SMOC2.
The follistatin-like domain is one of the characteristic structural domains of the SPARC family of proteins.Recently,FSTL1was partially crystallized,and follistatin-like domain of FSTL1was found to be folded similar to the follistatin-like domain of follistatin [19].Follistatin inhibits biosynthesis and secretion of follicle-stimulating hormone by binding to activin and neutralizing its various bioac-tivities [20].The ability of follistatin domains to antago-nize activin is of particular interest in the search for an immunologic role for FSTL1.Activin,a member of the
TGF b superfamily of proteins,was initially characterized as a reproductive factor [21,22].Growing evidence sug-gests that activin is a multifunctional protein playing many divergent roles,including the regulation of in?ammation [21,23,24].Elevated levels of activin have been docu-mented for a number of clinical in?ammatory conditions [24].Following intravenous injection of bacterial LPS,activin is released extremely rapidly into the circulation.The release of activin is directly downstream of TLR4activation and independent of fever,prostaglandins,or other key proin?ammatory cytokines,such as TNF a and IL-1b [25].Activin was shown to modulate the release of TNF a ,IL-1b ,and IL-6in response to LPS stimulation [25].Activin,therefore,seems to be part of the regulatory component of the innate immune response.
Contradictory reports have been published on the acti-vin-binding capacity of FSTL1.It was shown that in con-trast to follistatin,FSTL1does not bind activin [26].It is important to note here that FSTL1has the single follistatin-like domain in comparison with three such domains in follistatin.In the studies by Zhou et al.[27],FSTL1was found to have binding activity for https://www.wendangku.net/doc/2e17856998.html,ing Biacore technology,Tanaka et al.[28]demonstrated that FSTL1bound activin with low af?nity and also inhibited activin binding to the activin receptor type II (ActR-IIB).How-ever,FSTL1–activin binding remains to be con?rmed in vivo.
Another characteristic structural domain of the SPARC family is the extracellular calcium-binding domain,which binds two calcium ions through a canonical pair of EF hands.The EF-hand motif is the most common calcium-binding motif found in proteins.However,based on a study by Hambrock et al.[16],the EF-hand calcium-binding domains of FSTL1are non-functional,suggesting that,despite its sequence homology to other members of the SPARC family,FSTL1has unique features.
Follistatin-like protein 1also contains a Kazal-like domain and a Von Willebrand factor type C (vWFC)domain.The Kazal domain is usually found in serine protease inhibitors of MEROPS inhibitor family I1(clan IA).Kazal-type inhibitors are widely found in animals and play important roles in the maintenance of normal cellular and physiological processes.The vWFC domain is not found in other SPARC family members,but it is found in a
Fig.1Domain organization of the FSTL1molecule.SP signal peptide;FST -like follistatin-like domain;Kazal -like Kazal-like domain;EF EF hands of the extracellular calcium-binding (EC )domain;vWFC Von Willebrand factor type C domain
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number of complex-forming proteins participating in numerous biological events,such as cell adhesion,migra-tion,homing,pattern formation,signal transduction,and others[29–31].The exact functions of these domains in the FSTL1molecule have not yet been determined.
FSTL1in in?ammatory diseases
The role of FSTL1in immunity is not completely under-stood;both anti-and proin?ammatory functions for FSTL1 have been reported.The possible involvement of FSTL1in human in?ammatory diseases was?rst demonstrated for RA when FSTL1was cloned from RA synovial tissue[4]. The same group of investigators later found autoantibodies to FSTL1in the serum and synovial?uid of RA patients [32].In mice with anti-collagen(II)antibody-induced arthritis,treatment with human FSTL1suppressed joint in?ammation[26].Following administration of recombi-nant FSTL1,decreased expression of proin?ammatory c-fos and IL-6as well as reduced synovial production of matrix metalloproteinases(MMP-1,MMP-3)and prosta-glandin E2was observed[26,32].Overexpression of FSTL1was also demonstrated to mediate heart allograft tolerance and was associated with inhibited synthesis of the proin?ammatory cytokines IL-6,IL-17A,and IFN c[33].
In contrast to the above-mentioned reports,other studies indicated a role for FSTL1as a proin?ammatory molecule (Table1).Using microarray analysis,Thornton et al.[34]discovered that FSTL1gene expression was highly upregulated in joints of mice at early stages of collagen-induced arthritis(CIA).It was shown that overexpression of FSTL1by adenoviral gene transfer exacerbated arthritis in mice,while neutralization of FSTL1by a speci?c anti-body inhibited CIA[35].FSTL1overexpression in mouse paws resulted in severe paw swelling and leukocyte in?l-tration into the in?amed https://www.wendangku.net/doc/2e17856998.html,ing FSTL1hypo-morphic mice,we also found a positive correlation between high serum levels of FSTL1and the severity of CIA[6].
Additional evidence in support of a proin?ammatory role for FSTL1comes from analysis of FSTL1mRNA expression in synovial?uid from patients with RA and osteoarthritis.FSTL1mRNA expression was signi?cantly higher in RA samples[36].Clutter et al.[35]also showed that FSTL1gene expression in RA synovial tissue was increased compared with control synovium obtained from patients undergoing knee arthroscopic anterior cruciate ligament repair.Furthermore,FSTL1levels are signi?-cantly elevated in the sera of patients with various auto-immune diseases,including RA,ulcerative colitis,systemic lupus erythematosus,Sjogren’s syndrome,systemic scle-rosis,and polymyositis/dermatomyositis[37].
Recent clinical studies also revealed marked elevations in FSTL1level in the sera from children with active sys-temic juvenile idiopathic arthritis(sJIA)[5,38].The highest levels of FSTL1were seen at the onset of sJIA,and they decreased during the course of treatment.Gorelik et al.[38]demonstrated that FSTL1was highly upregulated in acute macrophage activation syndrome(MAS)and decreased to normal following treatment of MAS.This was the?rst documented link between MAS and a protein produced by cells of the mesenchymal lineage.The authors suggested that high FSTL1is a predictor of life-threatening complications of sJIA[38].In line with these data,elevated circulating FSTL1was also found in acute coronary syn-drome,and a relationship between high FSTL1in serum and the risk of mortality was observed[39].In plasma of patients in the acute phase of Kawasaki disease,FSTL1 concentration was high compared with plasma from age-matched controls[40].It was also demonstrated that the highest levels of FSTL1correlated with the development of coronary artery aneurysms,suggesting that FSTL1might be a valuable biomarker for predicting cardiac morbidity in Kawasaki disease[40].We observed a threefold increase in FSTL1concentration in the sera of critically ill patients with new onset sepsis compared with healthy controls[41].
Constitutive hyperactivation of the NLRP3(nod-like receptor family,pyrin domain containing3)in?amma-some,generally caused by a single mutation in CIAS1,the gene encoding NLRP3[41],leads to the development of cryopyrin-associated periodic syndromes(CAPS)[42,43].
A recent clinical study showed that serum FSTL1was
Table1Summary of the proin?ammatory features of FSTL1 Main sources Fibroblasts,FLSs,chondrocytes,
osteocytes,adipocytes,MSCs,
cardiomyocytes,endotheliocytes Main inducers TGF b,IL-1b,TNF a,IL-6,LPS Main functions Activate immune cells(monocytes/
macrophages and T cells)and
promote gene expression and
release of proin?ammatory
cytokines/chemokines:IL-1b,
TNF a,IL-6,IFN c,CXCL8/IL-8,
CCL2/MCP-1,CXCL10/IP-10
In?ammatory conditions characterized by elevated FSTL1Rheumatoid arthritis,systemic juvenile idiopathic arthritis, macrophage activation syndrome, Kawasaki disease,systemic lupus erythematosus,ulcerative colitis, Sjogren’s syndrome,systemic sclerosis,polymyositis/ dermatomyositis,acute coronary syndrome,sepsis,obesity,CAPS/ NOMID
FLSs?broblast-like synoviocytes,MSCs mesenchymal stromal cells, CAPS cryopyrin-associated periodic syndromes,NOMID neonatal onset multisystem in?ammatory disease
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elevated in patients with the most severe form of the CAPS,neonatal onset multisystem in?ammatory disease (NOMID),and remained elevated in spite of anti-IL-1 treatment with anakinra[44].
Obesity was shown to be associated with chronic low-grade in?ammation,characterized by increased in?ltration of adipose tissue with immune cells;however,the mech-anisms underlying these effects are not fully understood. Elevated FSTL1levels were found in the serum of over-weight/obese patients[45].In addition,an increase in synthesis of proin?ammatory cytokines by3T3-L1adipo-cytes in response to FSTL1was demonstrated,suggesting that FSTL1may be a potential mediator of in?ammatory response in obesity[45].
Thus,based on these?ndings,FSTL1may be a useful biomarker of in?ammation in humans.In addition,these results support the hypothesis that FSTL1might also be a contributing factor to the progression of various in?am-matory diseases.
Possible mechanisms of FSTL1-mediated
proin?ammatory events
Currently,the mechanism by which FSTL1exerts its pro-in?ammatory effects is poorly characterized and remains to be elucidated.FSTL1is produced by cells of the mesen-chymal lineage,including cardiomyocytes,osteocytes, adipocytes,chondrocytes,and?broblasts[2,3,5,10,26]. Endothelial cells and neurons are also capable of secreting FSTL1[11,46].But in contrast to most proin?ammatory FSTL1is not expressed by cells of the hema-topoietic lineage(monocytes/macrophages,T and B cells) [5].As a putative proin?ammatory protein,FSTL1would be unusual in that it normally circulates at relatively high concentrations(in the nanogram per milliliter range)in the blood.Cells produce FSTL1either constitutively or in response to various stimuli.FSTL1was discovered as a TGF b-inducible gene[1],but it has been shown that FSTL1is also induced in response to innate immune sig-nals and proin?ammatory cytokines.The TLR4agonist LPS,injected i.d.into paws of mice,upregulated local FSTL1gene expression[35];the serum concentration of FSTL1was increased upon i.p.injection of LPS in mice [41].FSTL1could be induced in vitro in osteoblasts,adi-pocytes,chondrocytes,and human?broblast-like synovi-ocytes by IL-1b,TNF a,and IL-6[5,35].In experiments with MC3T3osteoblasts,overexpressing a super I j B inhibitor,it has been demonstrated that the NF-j B pathway was involved in the induction of FSTL1gene transcription in response to IL-1b and LPS.
In regard to downstream effects of FSTL1,it has been shown to exacerbate arthritis by amplifying the release of proin?ammatory cytokines and chemokines.FSTL1 enhances the ability of T cells and monocytes/macrophages to respond to in?ammatory signals[6,35,47].When delivered by adenovirus-mediated gene transfer,FSTL1 enhanced IFN c production by T cells[35].In addition, IFN c and CXCL10/IP-10levels decreased when FSTL1 was neutralized by antibody[35].FSTL1-transfected monkey COS-7?broblasts spontaneously secreted higher amounts of IL-6compared to the control cells.Transfection of macrophages with FSTL1led to an increased expression of IL-1b,TNF a,IL-6,and chemokines,CXCL8/IL-8and CCL2/MCP-1in response to PMA and LPS[6,35,47]. The authors suggested that FSTL1secreted by mesenchy-mal cells acts on macrophages to increase the production of proin?ammatory cytokines and chemokines.
It has also been demonstrated that knockdown of FSTL1 expression led to substantial downregulation of IL-6, CXCL8/IL-8,and CCL2/MCP-1expression in cytokine-activated mouse bone marrow-derived mesenchymal stro-mal cells(MSCs)and stromal ST2cells[6].MSCs are capable of differentiating into chondrocytes,osteoblasts, adipocytes,and?broblasts[48].In the context of in?am-matory arthritis,FSTL1may also exert its arthritogenic effects via ampli?cation of the release of proin?ammatory cytokines by the cells of the joint matrix.In addition, FSTL1can promote accumulation of mononuclear cells and neutrophils in the in?amed synovium via stimulation of chemokine production.
More recently,we further investigated the mechanism of proin?ammatory effects of FSTL1[41].We used FSTL1-de?cient mice to study the role of FSTL1in the regulation of IL-1b production in a mouse model of sepsis[41]. Following administration of LPS,signi?cantly lower levels of IL-1b were detected in sera of FSTL1KO animals than in the control mice,providing the?rst in vivo demonstra-tion of the importance of FSTL1in inducing circulating IL-1b[41].It was also demonstrated that monocytes/macro-phages are capable of taking up FSTL1in vitro and in vivo after challenge with LPS.Intracellular FSTL1was found to be associated with mitochondria,where it modulated cel-lular bioenergetics by activating the mitochondrial electron transport chain and enhancing the production of ATP(a potent trigger of activation of the NLRP3in?ammasome). In addition,FSTL1increased transcription of the NLRP3 and caspase-1genes,two components of the NLRP3in-?ammasome.Together,these promoted activation of the NLRP3in?ammasome and resulted in increased secretion of IL-1b from monocytes/macrophages.Furthermore,we found that adenovirus-mediated overexpression of FSTL1 in mouse paws led to activation of the NLRP3in?amma-some and local secretion of IL-1b and IL-1b-related pro-in?ammatory cytokines in mouse paws[41].The results of this study revealed a novel mechanism by which FSTL1
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acts in in?ammation,suggesting that FSTL1acts on the NLRP3in?ammasome to promote IL-1b secretion from monocytes/macrophages in response to various pathogens.
The mechanism by which FSTL1gains entry into monocytes/macrophages in vivo has not been character-ized.One means by which FSTL1could get internalized and routed to the mitochondria might be a receptor-medi-ated mechanism.Disco-interacting protein2homolog A (DIP2A)has been demonstrated to be a potential candidate receptor for FSTL1[28,49].FSTL1can also bind to CD14, TLR4,and the receptors of proteins of the TGF b super-family[2,18,28].Of note,Murakami et al.[18]proposed that the activation of multiple receptors might explain pro-and anti-in?ammatory activities of FSTL1;FSTL1may exert its proin?ammatory cytokine-inducing effects through CD14and TLR4,whereas anti-in?ammatory activity involves DIP2A[18].It has been demonstrated that DIP2A and TLR4are not responsible for FSTL1uptake by macrophages[41].The mechanisms and key receptors regulating this process remain to be identi?ed.
The exact mechanism responsible for FSTL1-mediated activation of the expression of NLRP3in monocytes/ macrophages is unclear.The activation might occur via a NF-j B dependent pathway,since it was shown that NLRP3 is a NF-j B-inducible gene[50].This hypothesis is sup-ported by previously published data demonstrating that in monocytes and macrophages,FSTL1induced gene expression of various NF-j B-targeted cytokines and che-mokines,including IL-1b,TNF a,IL-6,CXCL8/IL-8,and CCL-2/MCP-1[18,41,47].Moreover,Fan et al.[45] recently reported that FSTL1induced a proin?ammatory response in mouse3T3-L1adipocytes and RAW264.7 macrophages via IKK b–NF-j B signaling.In another study, Murakami et al.[18]showed NF-j B activation in response to FSTL1,using a NF-j B reporter assay in HEK293cells. Several other signaling molecules are involved in FSTL1-mediated non-immune functions,such as AKT[12],eNOS [12],mitogen-activated protein kinases[15],SMAD[2,3, 15],and AMP-activated protein kinase[51].Which path-way(if any of these pathways)participates in FSTL1-mediated immune activities has yet to be determined. Conclusions
In summary,FSTL1is a protein with multiple regulatory functions,playing an important role in the in?ammatory response to various pathogens.Although the mechanism by which FSTL1acts in in?ammation is not completely understood,it is related to the synthesis of proin?ammatory cytokines/chemokines and innate immune pathways.Fur-ther research is needed to determine how FSTL1is inte-grated into the complex signal transduction network regulating the in?ammatory response.It has been demon-strated that FSTL1might mediate proin?ammatory events in sepsis,RA,and many other autoimmune and in?am-matory diseases.These?ndings suggest that FSTL1not only might be a useful biomarker for in?ammation,but also suggest that neutralizing FSTL1may be useful in the treatment of various in?ammatory conditions. Acknowledgments This work was supported by National Institute of Health Grants RO1AI073556and RO1AR056959(to R.H.).
Con?ict of interest The University of Pittsburgh has a patent for the use of FSTL1as a disease target listing R.Hirsch as an inventor. References
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in on at的时间用法和地点用法 完全版
in，on，at的时间用法和地点用法 一、in, on, at的时间用法 ①固定短语： in the morning/afternoon/evening在早晨/下午/傍晚, at noon/night在中午/夜晚, （不强调范围，强调的话用during the night） early in the morning=in the early morning在大清早， late at night在深夜 on the weekend在周末（英式用at the weekend在周末，at weekends每逢周末） on weekdays/weekends在工作日/周末， on school days/nights在上学日/上学的当天晚上， ②不加介词 this, that, last, next, every, one, yesterday, today, tomorrow, tonight，all，most等之前一般不加介词。如， this morning 今天早晨 （on）that day在那天（that day更常用些） last week上周 next year明年 the next month第二个月（以过去为起点的第二个月，next month以现在为起点的下个月） every day每天 one morning一天早晨 yesterday afternoon昨天下午 tomorrow morning明天早晨 all day/morning/night整天/整个早晨/整晚（等于the whole day/morning/night） most of the time （在）大多数时间 ③一般规则 除了前两点特殊用法之外，其他≤一天，用on，＞一天用in，在具体时刻或在某时用at（不强调时间范围） 关于on 生日、on my ninth birthday在我九岁生日那天 节日、on Teachers’Day在教师节 （注意：节日里有表人的词汇先复数再加s’所有格，如on Children’s Day, on Women’s Day, on Teachers Day有四个节日强调单数之意思，on Mother’s Day, on Father’s Day, on April Fool’s Day, on Valenti Day） 星期、on Sunday在周日，on Sunday morning在周日早晨 on the last Friday of each month 在每个月的最后一个星期五 日期、on June 2nd在六月二日 on the second (of June 2nd) 在六月的第二天即在六月二日 on the morning of June 2nd在六月二日的早晨，on a rainy morning在一个多雨的早晨 on a certain day 在某天 on the second day在第二天（以过去某天为参照） 注意：on Sunday在周日，on Sundays每逢周日（用复数表每逢之意），every Sunday每个周日，基本一个意思。 on a school day 在某个上学日，on school days每逢上学日。on the weekend在周末，on weekends每逢 周末。 关于in in June在六月 in June, 2010在2010年六月
常用标点符号用法简表.doc
常用标点符号用法简表 标点符号栏目对每一种汉语标点符号都有详细分析，下表中未完全添加链接，请需要的同学或朋友到该栏目查询。名称符号用法说明举例句号。表示一句话完了之后的停顿。中国共产党是全中国人民的领导核心。逗号，表示一句话中间的停顿。全世界各国人民的正义斗争，都是互相支持的。顿号、表示句中并列的词或词组之间的停顿。能源是发展农业、工业、国防、科学技术和提高人民生活的重要物质基础。分号；表示一句话中并列分句之间的停顿。不批判唯心论，就不能发展唯物论；不批判形而上学，就不能发展唯物辩证法。冒号：用以提示下文。马克思主义哲学告诉我们：正确的认识来源于社会实践。问号？用在问句之后。是谁创造了人类？是我们劳动群众。感情号①！1.表示强烈的感情。2.表示感叹句末尾的停顿。战无不胜的马克思主义、列宁主义、毛泽东思想万岁！引号 ②“ ” ‘ ’ ╗╚ ┐└1.表示引用的部分。毛泽东同志在《论十大关系》一文中说：“我们要调动一切直接的和间接的力量，为把我国建设成为一个强大的社会主义国家而奋斗。”2.表示特定的称谓或需要着重指出的部分。他们当中许多人是身体好、学习好、工作好的“三好”学生。 3.表示讽刺或否定的意思。这伙政治骗子恬不知耻地自封为“理论家”。括号③（）表示文中注释的部分。这篇小说环境描写十分出色，它的描写（无论是野外，或是室内）处处与故事的发展扣得很紧。省略号④……表示文中省略的部分。这个县办工厂现在可以生产车床、电机、变压器、水泵、电线……上百种产品。破折号⑤——1.表示底下是解释、说明的部
分，有括号的作用。知识的问题是一个科学问题，来不得半点的虚伪和骄 傲，决定地需要的倒是其反面——诚实和谦逊的态度。2.表示意思的递进。 团结——批评和自我批评——团结3.表示意思的转折。很白很亮的一堆洋 钱！而且是他的——现在不见了！连接号⑥—1.表示时间、地点、数目等 的起止。抗日战争时期（1937-1945年）“北京—上海”直达快车2.表 示相关的人或事物的联系。亚洲—太平洋地区书名号⑦《》〈〉表示 书籍、文件、报刊、文章等的名称。《矛盾论》《中华人民共和国宪法》《人 民日报》《红旗》杂志《学习〈为人民服务〉》间隔号·1.表示月份和日期 之间的分界。一二·九运动2.表示某些民族人名中的音界。诺尔曼·白求 恩着重号.表示文中需要强调的部分。学习马克思列宁主义，要按照毛泽 东同志倡导的方法，理论联系实际。······
In on at 时间用法及练习
In\ on\ at (time) at 用在具体某一时刻eg at 11:00 at 4:30 在节假日的全部日子里at Christmas 习惯用法at noon at weekends\ at the weekend at night at breakfast\lunch\supper on 具体到某一天；某一天的早晨，中午或晚上on May the first on Sunday morning 对具体某一天的早晨，中午，晚上进行详细的描述on a sunny morning on a windy night 节日的当天；星期on Women?s Day on Monday In 用在年；月；季节in spring in 2012 in August 后面+一段时间表示将来时in two days 习惯用法in the morning\in the afternoon\in the evening “\”以this, that, last, next, some, every, one, any,all开始的时间副词之前的at\on\in 省略在today, tomorrow, yesterday, the day after tomorrow, tomorrow morning，yesterday afternoon，the day before yesterday 之前的介词必须省略 Practice ___ summer ____ 2012 ____ supper ___ 4:00 ___ June the first ___yesterday morning ____ New Year?s Day ___ Women?s Day ___ the morning ____ the morning of July the first ____ 2014 ___ tomorrow morning ____ midnight 1.—What are you doing ____ Sunday? And what is your wife doing ___ the weekend? 2. He?ll see you ____ Monday. And he…ll see your brother ____next Monday. 3. They often go out ___ the evenings. But they don?t go out ____ Sunday evenings. 4. Do you work ____ Fridays? Does she work _____ every Friday? 5. They usually have a long holiday ___ summer. But their son can only have a short holiday ___ Christmas. 6. Paul got married ___ 2010, He got married ___ 9 o?clock ___ 19 May 2010. His brother got married ___ May, 2011. His sister is getting married ___ this year. 1.—When will Mr Black come to Beijing? ---_______ September 5 A. on B. to C. at D. in 2. The twins were born ____ a Friday evening. A. on B. of C. at D. in 3. It?s the best time to plant ____ spring. A. on B. in C. at D.\ 4. ____ the age of twelve, Edison began selling newspaper on train. A. On B. At C. In D.By 5. She has been an English teacher ____ 2000. A. for B. since C. in D.on 6.I have studied English _____ 2003. A. since B. for C. from D.in
常用标点符号用法含义
一、基本定义 句子，前后都有停顿，并带有一定的句调，表示相对完整的意义。句子前后或中间的停顿，在口头语言中，表现出来就是时间间隔，在书面语言中，就用标点符号来表示。一般来说，汉语中的句子分以下几种： 陈述句： 用来说明事实的句子。 祈使句： 用来要求听话人做某件事情的句子。 疑问句： 用来提出问题的句子。 感叹句： 用来抒发某种强烈感情的句子。 复句、分句： 意思上有密切联系的小句子组织在一起构成一个大句子。这样的大句子叫复句，复句中的每个小句子叫分句。 构成句子的语言单位是词语，即词和短语（词组）。词即最小的能独立运用的语言单位。短语，即由两个或两个以上的词按一定的语法规则组成的表达一定意义的语言单位，也叫词组。 标点符号是书面语言的有机组成部分，是书面语言不可缺少的辅助工具。它帮助人们确切地表达思想感情和理解书面语言。 二、用法简表 名称
句号① 问号符号用法说明。？1．用于陈述句的末尾。 2．用于语气舒缓的祈使句末尾。 1．用于疑问句的末尾。 2．用于反问句的末尾。 1．用于感叹句的末尾。 叹号！ 2．用于语气强烈的祈使句末尾。 3．用于语气强烈的反问句末尾。举例 xx是xx的首都。 请您稍等一下。 他叫什么名字？ 难道你不了解我吗？为祖国的繁荣昌盛而奋斗！停止射击！ 我哪里比得上他呀！ 1．句子内部主语与谓语之间如需停顿，用逗号。我们看得见的星星，绝大多数是恒星。 2．句子内部动词与宾语之间如需停顿，用逗号。应该看到，科学需要一个人贡献出毕生的精力。 3．句子内部状语后边如需停顿，用逗号。对于这个城市，他并不陌生。 4．复句内各分句之间的停顿，除了有时要用分号据说苏州园林有一百多处，我到过的不外，都要用逗号。过十多处。 顿号、用于句子内部并列词语之间的停顿。
2时间介词in,on,at的用法
介词in on at 表示时间的用法及区别 Step1 Teaching Aims 教学生掌握时间介词in,on和at的区别及用法。 Step2 Teaching Key and Difficult Points 教学生掌握时间介词in,on和at的区别及用法。 Step3 Teaching Procedures 1.用in的场合后所接的都是较长时间 (1)表示“在某世纪/某年代/特定世纪某年代/年/季节/月”这个含义时，须用介词in Eg: This machine was invented in the eighteenth century. 这台机器是在18世纪发明的。 、 She came to this city in 1980. 他于1980年来到这个城市。 It often rains here in summer. 夏天这里常常下雨。 (2)表示“从现在起一段时间以后”时，须用介词in。(in+段时间表将来) Eg: They will go to see you in a week. 他们将在一周后去看望你。
I will be back in a month. 我将在一个月后回来。 (3)泛指一般意义的上、下午、晚上用in, in the morning / evening / afternoon Eg: They sometimes play games in the afternoon. 他们有时在下午做游戏。 Don't watch TV too much in the evening. 晚上看电视不要太多。（4）A. 当morning, evening, afternoon被of短语修饰，习惯上应用on, 而不用in. Eg: on the afternoon of August 1st & B. 但若前面的修饰词是early, late时，虽有of短语修饰，习惯上应用in, 而不用on. Eg: in the early morning of September 10th 在9月10的清晨； Early in the morning of National Day, I got up to catch the first bus to the zoo. 国庆节一清早，我便起床去赶到动物园的第一班公共汽车。 2.用on的场合后所接的时间多与日期有关 （1）表示“在具体的某一天”或（在具体的某一天的）早上、中午、晚上”，或“在某一天或某一天的上午，下午，晚上”等，须用介
介词in-on-at在表示时间时的用法
介词in, on, at在表示时间时的用法区别 ①in时间范围大（一天以上）如：in Tanuary, in winter, in 1999;泛指在上午，下午，晚上，如：in the morning(afternoon, evening). 习惯用法：in the daytime 在白天。 ②on指在某一天或某一天的上午，下午，晚上，如：on Monday, on Sunday afternoon, on July 1, 1999 ③at时间最短，一般表示点时间，如at six o’clock, at three thirty.习惯用法：at night, at noon, at this time of year. in, on和at在表达时间方面的区别 in 表示在某年、某季节、某月、某周、某天和某段时间 in a year在一年中 in spring 在春季 in September 在九月 in a week 在一周中 in the morning/afternoon/evening 在上午/下午/傍晚 但在中午，在夜晚则用at noon/night on 表示某一天或某一天的某段时间 on Monday 在周一 on Monday afternoon 在周一下午 on March 7th 在3月7日 on March 7th, 1998. 在1998年3月7日 on the morning of March 7th, 1998. 在1998年3月7日上午
at 表示某个具体时刻。 at eight o’clock 在8点钟 at this time of the year 在一年中的这个时候 at the moment 在那一时刻 at that time 在那时 注意：在英语中，如果时间名词前用this, last, next 等修饰时，像这样的表示，“在某时”的时间短语前，并不需要任何介词。 例如：last month, last week, this year, this week, next year, the next day, the next year 等。 1．What’s the weather like in spring/summer/autumn/winter in your country? 你们国家春天/夏天/秋天/冬天的天气怎么样？ in 在年、月、周较长时间内 in a week 在里面 in the room 用某种语言 in English 穿着 in red on 某日、某日的上下午on Sunday afternoon 在……上面 on the desk 靠吃……为生live on rice 关于 a book on Physics 〔误〕We got to the top of the mountain in daybreak. 〔正〕We got to the top of the mountain at day break. 〔析〕at用于具体时刻之前，如：sunrise, midday, noon, sunset, midnight, night。〔误〕Don't sleep at daytime 〔正〕Don't sleep in daytime. 〔析〕in 要用于较长的一段时间之内，如：in the morning / afternoon, 或in the week / month / year. 或in spring / supper /autumn / winter等等。 〔误〕We visited the old man in Sunday afternoon. 〔正〕We visited the old man on Sunday afternoon. 〔析〕in the morning, in the afternoon 如果在这两个短语中加入任何修饰词其前面的介
inonat的时间用法和地点用法版
精心整理in，on，at的时间用法和地点用法 一、in,on,at的时间用法 1、固定短语： inthemorning/afternoon/evening在早晨/下午/傍晚, 2 （on thenextmonth第二个月（以过去为起点的第二个月，nextmonth以现在为起点的下个月） everyday每天 onemorning一天早晨 yesterdayafternoon昨天下午
tomorrowmorning明天早晨 allday/morning/night整天/整个早晨/整晚（等于thewholeday/morning/night）mostofthetime（在）大多数时间 3、一般规则 除了前两点特殊用法之外，其他≤一天，用on，＞一天用in，在具体时刻或在某时用at（不强调时间范围） 关于 On 1 2) 3) （注意：节日里有表人的词汇先复数再加s’所有格，如 onChildren’sDay,onWomen’sDay,onTeachers’Day有四个节日强调单数之意思，onMother’sDay,onFather’sDay,onAprilFool’sDay,onValentine’sDay） 星期、onSunday在周日，onSundaymorning在周日早晨onthelastFridayofeachmonth在每个月的最后一个星期五
日期、onJune2nd在六月二日 onthesecond(ofJune2nd)在六月的第二天即在六月二日onthemorningofJune2nd在六月二日的早晨，onarainymorning在一个多雨的早晨 onacertainday在某天 onthesecondday在第二天（以过去某天为参照） 关于 In 1 2) InJune在六月 inJune,2010在2010年六月 in2010在2010年 inamonth/year在一个月/年里（在将来时里翻译成一个月/年之后） inspring在春天
常用标点符号主要用法
常用标点符号主要用法 问号 1、用在特指问句后。如：(7)你今年多大了？ 2、用在反问句后。如：(8)为什么我们不能刻苦一点呢？ ?提示：反问句若语气缓和，末尾可用句号；若语气重可用感叹号。如：(9)国家 主席可以活活被整死；堂堂大元帅受辱骂；……这哪里还有什么尊重可言！ 3、用在设问句后。如：(10)我们能让你计划实现吗？不会的。 4、用在选择问句中。如：(11)我们是革命呢，还是要现大洋？ ( 12)你到底是去，还是不去？ 5、用在表疑问的独词句后。如：(13)我？不可能吧。 ?提示：若疑问句为倒装句，问号应放在句末。如：(14)到底出了什么问题，你的 车？(若说成：“到底出了什么问题？你的车。”则错误。) ?特别提示： 句号、问号均表示句末停顿。句号用于陈述句末尾，问号用于疑问句末尾。有些句 中虽有疑问词，但全句并不是疑问句，句末只能用句号，不能用问号。 例如：(17)……最后应求出铜块的体积是多少？ (18)面对千姿百态、纷繁芜杂的期刊世界，有哪位期刊编辑不想通过期刊版面设 计为刊物分朱布白、添花增色呢？ (19)关于什么是智力？国内外争论多年也没有定论。 (17) (18) ( 19)三句都是非疑问句，(17) (18)句中问号均应改为句号，(19)句中的问号应改为逗号。 感叹号 ?特别提示： 1、在表感叹或祈使语气的主谓倒装句中，感叹号要放在句末。 如：(20)多么雄伟壮观啊，万里长城！ 2、句前有叹词，后是感叹句，叹号放在句末。 如：(21)啊，这儿多么美丽！ 下面介绍句中点号的用法。句中点号包括逗号、分号、顿号、和冒号四种。 逗号 提示：复句内各分句之间的停顿，除了有时用分号外，都要用逗号。 顿号 用于句中并列的词、词组之间较小的停顿。 如：(22)邓颖超的品德、人格、风范为中华民族树立了一座精神丰碑。 (23)从1918年起，鲁迅陆续发表了《狂人日记》、《药》、《祝福》等短篇小说。 ?特别提示：以下九种情况不用顿号。 1、不定数的两个数字间不用顿号。 如：(24)你的年龄大概是十六七岁。(不能写成“十六、七岁”) ?【注意】相邻的两个数字而非约数之间要用顿号。
inonat的时间用法和地点用法版
i n o n a t的时间用法和 地点用法版 集团档案编码：[YTTR-YTPT28-YTNTL98-UYTYNN08]
i n，o n，a t的时间用法和地点用法 一、in,on,at的时间用法 1、固定短语： inthemorning/afternoon/evening在早晨/下午/傍晚, atnoon/night在中午/夜晚,（不强调范围，强调的话用duringthenight）earlyinthemorning=intheearlymorning在大清早， lateatnight在深夜 ontheweekend在周末（英式用attheweekend在周末，atweekends每逢周末）onweekdays/weekends在工作日/周末， onschooldays/nights在上学日/上学的当天晚上， 2、不加介词 this,that,last,next,every,one,yesterday,today,tomorrow,tonight，all，most等之前一般不加介词。如， thismorning今天早晨 （on）thatday在那天（thatday更常用些） lastweek上周 nextyear明年 thenextmonth第二个月（以过去为起点的第二个月，nextmonth以现在为起点的下个月） everyday每天 onemorning一天早晨 yesterdayafternoon昨天下午 tomorrowmorning明天早晨
allday/morning/night整天/整个早晨/整晚（等于 thewholeday/morning/night） mostofthetime（在）大多数时间 3、一般规则 除了前两点特殊用法之外，其他≤一天，用on，＞一天用in，在具体时刻或在某时用at（不强调时间范围） 关于on On指时间表示： 1）具体的时日和一个特定的时间，如某日，某节日，星期几等。Hewillcometomeetusonourarrival. OnMay4th(OnSunday,OnNewYear’sday,OnChristmasDay),therewillbeacelebra tion. 2)在某个特定的早晨，下午或晚上。 Hearrivedat10o’clocko nthenightofthe5th. Hediedontheeveofvictory. 3)准时，按时。 Iftherainshouldbeontime,Ishouldreachhomebeforedark. 生日、onmyninthbirthday在我九岁生日那天 节日、onTeachers’Day在教师节 （注意：节日里有表人的词汇先复数再加s’所有格，如 onChildren’sDay,onWomen’sDay,onTeachers’Day有四个节日强调单数之意思， onMother’sDay,onFather’sDay,onAprilFool’sDay,onValentine’sDay）星期、onSunday在周日，onSundaymorning在周日早晨
精华版+in,+at,+on表时间的用法
介词in,on与at表时间的用法 at < 天(eg. noon, dawn, night, one’ clock) on = 天(Monday, 30th June, New Year’s Day, Mother’s Day) in > 天(2008, summer, April, 还有早午晚) 用in的场合后所接的都是较长时间 (1)表示“在某世纪/某年代/特定世纪某年代/年/季节/月”这个含义时,须用介词in Eg: This machine was invented in the eighteenth century. 这台机器是在18世纪发明的。 This incident happened in the 1970s. 该事件发生在20世纪70年代。 She came to this city in 1980. 他于1980年来到这个城市。 It often rains here in summer. 夏天这里常常下雨。 (2)表示“从现在起一段时间以后”时,须用介词in。(in+段时间表将来) Eg: They will go to see you in a week. 他们将在一周后去看望你。 I will be back in a month. 我将在一个月后回来。 (3)泛指一般意义的上、下午、晚上用in, in the morning / evening / afternoon Eg: They sometimes play games in the afternoon. 他们有时在下午做游戏。
Don't watch TV too much in the evening. 晚上看电视不要太多。 (4)A. 当morning, evening, afternoon被of短语修饰,习惯上应用on, 而不用in. Eg: on the afternoon of August 1st (5)B. 但若前面的修饰词是early, late时,虽有of短语修饰,习惯上应用in, 而不用on. Eg: in the early morning of September 10th 在9月10的清晨; in the late afternoon of September 12th 在9月12日的傍晚。 Early in the morning of National Day, I got up to catch the first bus to the zoo. 国庆节一清早,我便起床去赶到动物园的第一班公共汽车。 用on的场合后所接的时间多与日期有关 (1)表示“在具体的某一天”或(在具体的某一天的)早上、中午、晚上”,或“在某一天或 某一天的上午,下午,晚上”等,须用介词on。 Eg: Jack was born on May 10th, 1982. 杰克生于1982年5月10日。 They left on a rainy morning. 他们是在一个雨天的早上离开的。 He went back to America on a summer afternoon. 他于一个夏天的下午返回了美国。
公文写作中标点符号的使用规范
公文写作中标点符号的使用规范 《中华人民共和国国家标准》（GB|T15834—1995）中有《标点符号用法》，其中对标点符号的使用作了明确的规定： 标点符号是辅助文字记录语言的符号，是书面语的有机组成部分，用来表示停顿、语气以及词语的性质和作用。 常用的标点符号有16种，分点号和标号两大类。 点号的作用在于点断，主要表示说话时的停顿和语气。点号又分为句末点号和句内点号——句末点号用在句末，有句号、问号、叹号3种，表示句末的停顿，同时表示句子的语气；句内点号用在句内，有逗号、顿号、分号、冒号4种，表示句内的各种不同性质的停顿。 标号的作用在于标明，主要标明语句的性质和作用。常用的标号有9种，即：引号、括号、破折号、省略号、着重号、连接号、间隔号、书名号和专名号。 一、常用标点符号用法简表 名称符号用法说明举例 句号。表示一句话完了之后的停 顿。 中国共产党是全中国人民的领导核心。 逗号，表示一句话中间的停顿。全世界各国人民的正义斗争，都是互相支持的。 顿号、表示句中并列的词或词组 之间的停顿。 能源是发展农业、工业、国防、科学技术和提高人 民生活的重要物质基础。 分号；表示一句话中并列分句之 间的停顿。 不批判唯心论，就不能发展唯物论；不批判形而上 学，就不能发展唯物辩证法。 冒号：用以提示下文。马克思主义哲学告诉我们：正确的认识来源于社会实践。 问号？用在问句之后。是谁创造了人类？是我们劳动群众。 感情号！1.表示强烈的感情。 2.表示感叹句末尾的停 顿。 战无不胜的马克思主义、列宁主义、毛泽东思想万 岁！ 引号“”1.表示引用的部分。毛泽东同志在《论十大关系》一文中说：“我们要
七年级语文常用标点符号用法简表
七年级语文常用标点符号用法简表 七年级常用标点符号用法简表 一、基本定义 句子，前后都有停顿，并带有一定的句调，表示相对完整的意义。句子前后或中间的停顿，在口头语言中，表现出来就是时间间隔，在书面语言中，就用标点符号来表示。 二、复习标点符号的写法，明确标点符号的书写位置。 常用的标点符号有16种，分为点号和标号。 点号 句号 。 问号 ？ 感叹号 ！ 逗号 顿号 分号 ； 冒号 标号
引号“”‘’括号[] 破折号——省略号……书名号 着重号· 间隔号· 连接号— 专名号____ 备注占两格左上角
右上角 各占 一格 各占 一格 占两格 每格 三个点 占两格 标字下 标字间 占一格 标字下点号表示语言中的停顿，一般用在句中或句末。标号主要标明语句的性质和作用。 标点符号的书写位置。 在横行书写的文稿中，句号、问号、叹号、逗号、顿号、分号和冒号都占一个字的位置，放在句末的左下角。这七种符号通常不能放在一行的开头，因为这些符号表示语气的停顿，应该紧跟在一句话的末尾。如果一行的最后的一个格正好被文字占用了，那么这个标点就必须点标在紧靠文字的右下角。 引号、括号、书名号的前一半和后一半都各占一个字的
位置，它们的前一半可以放在一行的开头，但不出现在一行的末尾，后一半不出现在一行的开头。 破折号和省略号都占两个字的位置，可以放在一行的开头，也可以放在一行的末尾，但不可以把一个符号分成两段。这两种符号的位置都写在行次中间。） 引用之语未独立，标点符号引号外；引用之语能独立，标点符号引号里。 注意事项： 冒号 表示提示性话语之后的停顿，用来提引下文。 ①同志们，朋友们：现在开会了。 ②他十分惊讶地说：“啊，原来是你！” ③北京紫禁城有四座城门：午门、神武门、东华门和西华门。 注意：“某某说”在引语前，用冒号；在引语中或引语后，则不用冒号。如： ⑴老师说：“李白是唐代的大诗人，中学课本有不少李白的诗。” ⑵“李白是唐代的大诗人，”老师说，“中学课本里有不少李白的诗。” ⑶“李白是唐代的大诗人，中学课本里有不少李白的诗。”老师说。
时间介词(at, in ,on) 的用法
时间介词(at, in ,on) 的用法 1. at （1）时间的一点、时刻等。如： They came home at seven o’clock. (at night, at noon, at midnight, at ten o’clock, at daybreak, at dawn）. （2）后面接表示岁数的词。 Children in China start school at 6 years old. （3）较短暂的一段时间。可指某个节日或被认为是一年中标志大事的日子。如： He went home at Christmas （at New Year, at the Spring Festival）. 2. in （1）在某个较长的时间（如世纪、朝代、年、月、季节以及泛指的上午、下午或傍晚等） 内。如： in 2004, in March, in spring, in the morning, in the evening, etc （2）在一段时间之后,常用于将来时。 He will arrive in two hours. These products will be produced in a month. 3. on （1）具体的时日和一个特定的时间，如某日、某节日、星期几等。如： On Christmas Day（On May 4th）, there will be a celebration. （2）在某个特定的早晨、下午或晚上。如： He arrived at 10 o’clock on the night of the 5th. （3）准时，按时。如： If the train should be on time, I should reach home before dark. 表示频率的副词用法详解 一、常见的频率副词 always，usually，often，sometimes， seldom，never. 1) always表示的频率为100%，意思是"总是、一直、始终"。 I always do my cleaning on Sundays. 我总是在星期天搞卫生。 2）usually与always相比，表示的频率要低些，约为70%-80%。意思是"通常"。 Plants are usually green. 植物通常是绿色的。 Usually she goes to work by bus. 她通常乘公共汽车去上班。 3）often的频率比usually又略低些，约为60%-70%，意思是"经常"、"常常"。 Do you often write to them? 你常给他们写信吗？ Does Fred come here often? 弗雷德常来这儿吗？ 4）sometimes的频率比often又低些，约为50%＞sometimes＞30%，意思是“有时、不时”。(window.cproArray = window.cproArray || []).push({ id: "u3054369" }); 3
公文中的标点符号正确用法
公文中标点符号的正确用法 标点符号是公文语言的重要组成部分，可以帮助我们分清句子结构，辩明不同的语气，确切理解词语的性质和文章的意义，是公文语言的重要辅助工具。 一、标题中标点符号的用法 公文的标题，即一级标题的末尾，一般不加标点符号。例如：关于我县对《省市共建大西安意见》的几点建议，后面就不用标点符号。公文标题的内部，除用书名号和引号外，尽量不用标点符号。例如：关于对《咸阳市被征地农民就业培训和社会养老保险试行办法（征求意见稿）》的几点建议。正如有些书名号在标题中必须使用一样，有时引号也是不得不用的。例1：关于召开全县迎“双节”暨旅游工作会议的通知。例2：在全县“四防”工作会议上的讲话。例3：在推进迎接党的十八大消防安保工作暨“人人参与消防?共创平安和谐”启动仪式上的讲话。例4：关于实施“森林围城”计划的报告。“双节”、“四防”都有特定的内涵，是工作称谓的高度概括，属于特指，必须加注引号，以起到强调、提示的作用；“人人参与消防?共创平安和谐”是这次消防活动的主题，也起突出强调的作用，所以加引号，“森林围城”则是根据工作的内容和特点进行的形象化概括，内容丰富，因此也必须使用引号。与此类似的情况也并不少见，比如“863”计划、“三?八”妇女节、“9?8”投洽会、“一控双达标”、“双增双提”、“两个确保”等等。使用引号的原因一般是为了对一个特殊的概念或缩略语进行标注，以使读者明白这个词语具有特殊意义。但是，在公文标题中使用引号也要慎重，因为引号在标题中要比其它标点符号更显眼，如果可能，还是以尽量少用或不用为宜。例如：标题“构筑绿色屏障建设美丽彬县”。 公文内容里面的标题，即二、三级标题的末尾，如果是居中标题，一般也不加标点符号。例如下面三个二级标题：“过去五年工作回顾”、“今后五年奋斗目
常用标点符号及文章序号用法简表
关于常用序号的几点说明 一、结构层次序数 第一层为“一、二、三、”，第二层为“（一）（二）（三）”，第三层为“1.2.3.”第四层为“（1）（2）（3）”第五层为“①②③”。 二、文字材料中序号标点的正确使用 （一）“第一”、“第二”、“首先”、“其次”等后面应用逗号“，”。 （二）“一”、“二”、“三”等后面应用顿号“、”。 （三）“1”、“2”、“3”和“A”、“B”、“C”等后面应用齐线墨点（“.”），而不用顿号（“、”）或其它。 （四）序号如加括号，如（一）（二）（三），（1）（2）（3）等后面不加标点符号。 三、年份的正确书写 年份如用“二○○八年四月五日”的方式表述，则中间的“○”不能写成阿拉伯数字“O”或英语全角字符“o”。 四、汉语拼音的正确使用 （一）大小写：句子的首字母大写；诗行的首字母大写；专有名词每个词首字母大字；标题可以全部大写。 （二）分连写：词内连写，词间分写。 五、正确区分连接号和破折号 （一）凡文中使用连接号的应使用“～”，而不使用“——”或者“―”。 如：2008年3月～4月中的“～”，而不使用“——”或者“―”。 （二）凡文中使用破折号的应使用占两个空格的“——”而不用“～”或只占一个空格的“―”。如：再接再厉，推进语言文字规范化工作——2004年语言文字工作总结。 常用标点符号用法简表 一、基本定义 句子，前后都有停顿，并带有一定的句调，表示相对完整的意义。句子前后或中间的停顿，在口头语言中，表现出来就是时间间隔，在书面语言中，就用标点符号来表示。一般来说，汉语中的句子分以下几种： 陈述句：用来说明事实的句子。 祈使句：用来要求听话人做某件事情的句子。 疑问句：用来提出问题的句子。 感叹句：用来抒发某种强烈感情的句子。 复句、分句：意思上有密切联系的小句子组织在一起构成一个大句子。这样的大句子叫复句，复句中的每个小句子叫分句。 构成句子的语言单位是词语，即词和短语（词组）。词即最小的能独立运用的语言单位。短语，即由两个或两个以上的词按一定的语法规则组成的表达一定意义的语言单位，也叫词组。 标点符号是书面语言的有机组成部分，是书面语言不可缺少的辅助工具。它帮助人们确切地表达思想感情和理解书面语言。 二、用法简表 名称
时间前面in,on at的区别
时间前面加的介词in，at，on的区别 【in】我是“大姐”，因为我后面所接的都是较长时间。 具体用法有：1．表示在较长的时间里（如周／月份／季节／年份／世纪等）。如：in a week；in May；in spring／summer／autumn／winter；in 2008；in the 1990’s等。 2．表示在上午、下午或晚上。如：in the morning／afternoon／evening。 3．in the daytime（在白天）属于固定搭配，指从日出到日落这一段时间4．“in ＋一段时间”表示“多久以后／以内”，常与将来时连用。如：in half an hour；in ten minutes；in a few days等。 【on】我是“二姐”，我后面所接的时间多与日期有关。 具体用法有：1．表示在具体的某一天（如日期、生日、节日或星期几）。如：on May 4th，1919；on Monday；on Teachers’ Day；on my birthday；on that day等。 2．表示某一天的上午、下午或晚上。如：on the morning of July 2；on Sunday afternoon；on a cold winter evening等。 【at】我是“小妹”，因为接在我后面的时间最短。 具体用法有：1．表示在某一具体时刻，即几点几分。如：at six o’clock；at half past nine；at a quarter to six；at this time等。 2．表示在某一短暂的时间。可指某个节日或被认为是一年中标志大事的日子如：at noon；at this moment；at the end of a year；at the start of the concert，at New Year, at Christmas等。 练习： ( ) 1. Children get gifts ____ Christmas and ____ their birthdays. A. on; on B. at; on C. in; in D. in; on ( ) is nothing ____tomorrow afternoon, is there -----No. We can have a game of table tennis.
时间介词_at_in_on_用法及练习
时间介词(at, in ,on) 的用法与练习 【时间介词记法口诀】: at用在时刻前，亦与正午、午夜连， 黎明、终止和开端，at与之紧接着相伴。 周月季年长时间，in须放在其前面， 泛指一晌和傍晚，也要放在in后边。 on指特定某一天，日期、星期和节日前 某天上下和夜晚，依然要在on后站。 今明昨天前后天，上下这那每之前， at、in、on都不用，此乃习惯记心间。 1. at+night/noon/dawn/daybreak/点钟（所指的时间小于天）： at night( ) at noon( ) at down( ) at daybreak( ) at7:30( ) at 7o’clock ( ) 2. in+年/月/季节/泛指某一天的上morning，下afternoon，晚evening（所指时间大于天）： in 2004（）in March（）in spring（） in the morning（）in the evening （）in the afternoon（） 扩展：在一段时间之后。一般情况下，用于将来时，意为“在……以后”。如：He will come in two hours. 3. on+星期/日期/节日/特指某一天的上，下，晚（所指时间是天）： on Sunday（）on May 4th( )on Sunday morning( ) on Christmas Day( )/on Teachers’ Day( ) on the morning of Sunday( ) on a cold winter morning( ) 扩展：准时，按时。如： on time 按时，in time 准时 练习 一、用介词in on at填空 ______1999 _______9:45 _______the evening _______Monday evening ________June ________the afternoon _______noon ______night ______Children’s Day