005Graphene on Pt(111)_Growth and substrate interaction

生态毒理学报Asian Journal of Ecotoxicology第19卷第2期2024年4月V ol.19,No.2Apr.2024㊀㊀基金项目:国家自然科学基金项目(31960154);中央引导地方科技发展资金项目(2023ZY0004);内蒙古自治区 草原英才 工程青年创新创业人才项目(Q2022085);内蒙古自治区高等学校科学研究项目(NJZZ23017);内蒙古自治区自然科学基金项目(2023QN03047);内蒙古医科大学面上项目(YKD2022MS033)㊀㊀第一作者:王惠增(1997 ),女,硕士研究生,研究方向为生殖毒理㊁分子诊断,E -mail:******************* ㊀㊀*通信作者(Corresponding author ),E -mail:*********************.cnDOI:10.7524/AJE.1673-5897.20230413002王惠增,刘秉春,陈红,等.苯并[a]芘对生殖系统的毒性作用及其机制研究进展[J].生态毒理学报,2024,19(2):165-183Wang H Z,Liu B C,Chen H,et al.Research progress on toxic effects of benzo(a)pyrene on reproductive system and its mechanism [J].Asian Journal of Ecotoxicology,2024,19(2):165-183(in Chinese)苯并[a ]芘对生殖系统的毒性作用及其机制研究进展王惠增1,刘秉春2,陈红1,徐沛欣1,郭鑫1,袁建龙1,*1.内蒙古医科大学附属医院检验科,呼和浩特0100502.内蒙古医科大学附属医院干细胞实验室/内蒙古自治区肿瘤细胞基因检测应用与研究工程实验室,呼和浩特010050收稿日期:2023-04-13㊀㊀录用日期:2023-11-02摘要:苯并[a]芘(benzo(a)pyrene,BaP)作为多环芳烃(polycyclic aromatic hydrocarbons,PAHs)的成员,是最早发现也是最具有代表性的环境污染物,通过空气㊁食物㊁水源等途径进入人体,引起细胞氧化应激损伤㊁DNA 损伤和基因异常表达导致细胞死亡㊂研究表明雄性与雌性动物经BaP 染毒后,其生殖器官㊁生殖细胞甚至激素水平均会受到影响,进而影响受精卵形成和胚胎发育,造成不良妊娠结局㊂因此,近年来BaP 的生殖毒性受到广泛关注,其作用机制包括改变胞内活性氧水平㊁诱导细胞DNA 损伤以及调控生殖发育相关基因㊁类固醇合成相关基因和促凋亡基因影响生殖发育㊂BaP 作为环境毒物,不仅可以影响生态环境的稳定性,还可以影响生物的生殖发育,损害生态环境中的物种多样性,从长远来看,BaP 的不良影响不但会威胁到陆地与海洋生物种群的稳定,还会破坏陆地和海洋生态系统的功能㊂本文将从生殖健康㊁配子与合子形成以及胚胎发育的角度,详细阐述BaP 染毒对生殖系统的毒性作用与机制,为预防BaP 引起的生殖危害㊁减少不良妊娠结局提供理论依据,旨在为BaP 的环境毒性行为和对生物的毒性研究提供有效借鉴,为合理预防和缓解因接触BaP 等环境毒物而带来的健康影响提供参考㊂关键词:苯并[a]芘(BaP);生殖细胞;生殖毒性;生殖器官;激素;细胞毒性文章编号:1673-5897(2024)2-165-19㊀㊀中图分类号:X171.5㊀㊀文献标识码:AResearch Progress on Toxic Effects of Benzo (a )pyrene on Reproductive System and Its MechanismWang Huizeng 1,Liu Bingchun 2,Chen Hong 1,Xu Peixin 1,Guo Xin 1,Yuan Jianlong 1,*1.Department of Laboratory Medicine,The Affiliated Hospital of Inner Mongolia Medical University,Hohhot 010050,China2.Stem Cell Research Center,The Affiliated Hospital of Inner Mongolia Medical University/Inner Mongolia Autonomous Region Tumor Cell Gene Detection Application and Research Engineering Laboratory,Hohhot 010050,ChinaReceived 13April 2023㊀㊀accepted 2November 2023Abstract :Benzo(a)pyrene (BaP),as a member of the polycyclic aromatic hydrocarbons (PAHs),is the earliest dis -covered and most representative environmental pollutant.It enters the human body through the air,food,and water,causing cellular oxidative stress damage,DNA damage,and abnormal gene expression,leading to cell death.Studies have shown that when male and female animals are exposed to BaP,their reproductive organs,cells,and hormone166㊀生态毒理学报第19卷levels are affected,which in turn will affect the formation of fertilized eggs and embryonic development,resulting in adverse pregnancy outcomes.Hence,the reproductive toxicity of BaP has received more attention in recent years.Its mechanism of action on reproductive development includes alteration of intracellular reactive oxygen species levels,induction of cellular DNA damage,and modulation of genetic changes related to reproductive development,steroid synthesis and pro-apoptosis.BaP,as an environmental toxicant,could influence the stability of the ecological environment,the reproductive development of organisms and destroy the diversity of species in the ecosystems.In this review,we will detailly elaborate on the toxic effects and mechanisms of BaP on the reproductive system,and provide a theoretical evidence for prevention reproductive harm caused by BaP and the reduction of adverse pregnancy outcomes,with the aim to providing an effective reference for the study of BaP s toxicity to the environment and organisms,and for the rational prevention and mitigation of the health effects of exposure to BaP or other environmental toxins.Keywords:benzo(a)pyrene;germ cell;reproductive toxicity;genital organ;hormone;cytotoxicity㊀㊀PAHs是由2个或2个以上的稠环芳烃组成的有机化合物[1],由于其化学性质稳定且具有疏水性[2],因此多环芳烃可以在环境中稳定存在,是常见的环境污染物,广泛存在于油炸烧烤食物㊁香烟烟雾[3]㊁汽车尾气[4]㊁煤炭燃烧[5]等中㊂人类可以通过空气㊁饮用水㊁食物等不同方式暴露于多环芳烃[6]㊂此外,多环芳烃的亲脂性有利于它们在水生生物的脂肪中积累[7],并随着食物链进入人体,对人类健康产生威胁㊂BaP是多环芳烃中最具有代表性也是毒性最大的致癌物[8],可以诱发肺癌[9]㊁乳腺癌[10]等癌症,危害人类健康㊂BaP广泛存在于人类生活环境中,2019年公布的美国毒物和疾病登记机构物质优先清单中,BaP被列为第8名,在污染的空气[11]㊁土壤[12]㊁水源[13]㊁食物[14]中均可以检测到BaP㊂近年来,越来越多研究表明BaP与胚胎畸形[15]和不良妊娠[16]有着密切的联系㊂在妊娠早期暴露于BaP会导致小鼠胎儿畸形率增高[17]㊂此外,一项病例对照研究表明,接触BaP与早孕流产之间存在联系,妊娠女性发生流产的风险与血中BaP-DNA加合物的浓度成正比,这进一步说明BaP除了致癌性也具有生殖毒性㊂目前对于BaP的研究多聚焦于其诱发癌症[18-19]尤其是肺癌[20]这一方面,虽有研究表明BaP 具有生殖毒性,其生殖毒性机理尚未研究透彻㊂本综述的目的是总结BaP生殖毒性相关文章,讨论BaP导致生殖毒性的潜在分子机制㊂1㊀BaP在生殖方面的主要致毒途径(The main toxic pathway of BaP in reproduction)近些年研究发现,BaP发挥其致毒作用主要有3种途径:(1)通过氧化应激影响细胞正常代谢;(2)BaP可以与DNA形成加合物,进而导致DNA损伤;(3)BaP可以通过调控基因表达,发挥其毒性作用㊂BaP致毒途径是多种机制相辅相成㊂由于生殖对繁育后代具有重要意义,因此研究BaP的生殖毒性已成为科学家们的研究重点,下文将重点总结BaP的生殖毒性机制㊂1.1㊀氧化应激(Oxidative stress)BaP进入细胞后,通过AHR途径诱导细胞发生氧化应激反应,其主要过程为:BaP刺激细胞质中的一种转录因子 芳香族化合物受体(aryl hydrocar-bon receptor,AHR)[21],使其转入到细胞核后,再与芳香族化合物受体核转运蛋白(aryl hydrocarbon recep-tor nuclear transporter,ARNT)结合形成异二聚体[22],结合在下游靶基因上,激活细胞色素P450目标基因的异常表达,包括细胞色素P4501A1(cytochrome P450family1subfamily A member1,CYP1A1)㊁细胞色素P4501A1(cytochrome P450family1subfamily A member2,CYP1A2)㊁细胞色素P4501B1(cyto-chrome P450family1subfamily B member1, CYP1B1)[21,23],进而引起细胞产生大量活性氧(reac-tive oxygen species,ROS),使机体发生氧化应激反应,如果体内的活性氧产生过多,超出了细胞的清除能力,会影响细胞的正常代谢甚至会破坏细胞结构㊂低㊁高剂量的BaP均可导致小鼠卵母细胞功能障碍,降低精卵结合与融合率,这与线粒体ROS水平增加和卵膜脂质过氧化密切相关[24]㊂Zhang等[25]发现BaP可以削弱雌鼠的繁殖能力,通过增加雌鼠卵母细胞中ROS,扰乱纺锤体组装,染色体配对,阻滞卵母细胞减数分裂过程㊂BaP诱导的氧化应激不仅仅通过产生ROS这一条途径,还可以通过降低过氧第2期王惠增等:苯并[a]芘对生殖系统的毒性作用及其机制研究进展167㊀化氢酶(catalase,CAT)㊁抗坏血酸过氧化物酶(ascor-bate peroxidase,AP)㊁谷胱甘肽过氧化物酶(glutathione peroxidase,GPX)㊁超氧化物歧化酶(superoxide dis-mutase,SOD)㊁谷胱甘肽还原酶(glutathione reductase, GR)等抗氧化酶的活性[26-27]以及促进炎症细胞因子表达[28]导致氧化应激的发生,最终引起细胞功能受损㊂1.2㊀BPDE引起DNA损伤(BPDE induces DNA damage)BaP进入体内经过一系列氧化代谢反应,生成二羟环氧苯并[a]芘(BaP-7,8-dihydrodiol-9,10-epoxide, BPDE),进而发挥其毒性,Penning[29]认为生成BPDE 的主要途径是在细胞色素P450酶的催化下,BaP末端的苯环上发生单加氧化反应,生成BaP-7,8-环氧化物(BaP-7,8epoxide),在环氧化物水解酶作用下转化为BaP-7,8-二氢二醇(BaP-7,8diol),该过程循环往复最终形成致癌物 BPDE[30-32]㊂BPDE可以与DNA共价结合形成加合物,造成DNA损伤㊂Shiizaki等[33]提出一个关于BaP-DNA加合物成因的假设,即CYP1A1是BaP被激活形成BPDE反应中的关键酶,这与Bukowska等[32]提出的观点一致㊂Einaudi等[34]通过建立BaP染毒的雌性小鼠模型,发现BaP可以导致卵母细胞与卵丘细胞DNA损伤,并且他们认为导致DNA断裂的主要原因是由于细胞中的修复机制对BPDE-DNA加合物切除和修复导致的㊂Zhan等[35]研究表明BaP形成的DNA加合物可以干扰DNA复制,进一步引起胚胎的DNA损伤,影响胚胎的发育㊂Zhan等[35]进一步研究发现DNA加合物与ROS共同造成基因组严重损伤,还可以引起卵裂球的端粒功能障碍,最终引起胚胎的异常㊂Miao等[36]发现BaP会引起猪卵母细胞纺锤体组装缺陷进一步引起减数分裂停滞,而导致这一结果的原因可能是DNA加合物引起的㊂Zhang 等[25]将小鼠卵母细胞暴露于BaP后,发现纺锤体的组装㊁染色体的排列和着丝点-微管附着均被破坏,这可能与DNA加合物的形成有关联,与Miao的设想一致㊂1.3㊀基因表达调控(Regulation of gene expression)基因表达调控是生物学研究的重要内容之一,在细胞分化发育的不同时期,基因表达的种类和强度各不相同,共同决定着细胞的形态与功能;细胞为了适应环境变化改变自身的基因表达有利于生存,因而基因表达调控十分重要㊂海洋污染问题日趋严重,BaP具有水生生物生殖毒性,是造成海洋污染的重要原因之一,受到广泛关注㊂有研究发现BaP生殖毒性的潜在分子机制是通过调控相关基因表达㊂数字基因表达技术表明BaP对雄性栉孔扇贝睾丸中的生殖基因有影响,其中热休克蛋白90㊁细胞色素P4503A㊁凋亡抑制蛋白3个基因的改变会引起睾丸组织损伤,此外BaP与性激素合成和睾丸发育相关基因有密切联系[37]㊂Albornoz-Abud等[38]研究表明苯并芘可以通过调控GH/IGF轴发挥其生殖毒性,急性暴露于BaP会导致尼罗罗非鱼睾丸中内分泌相关基因:胰岛素样生长因子1(insulin-likegrowth factor1,IGF1)和生长激素受体基因1(growth hormone receptor1,GHR1)基因表达降低,并造成发育问题㊂BaP通过基因调控引起的生殖毒性不仅仅在海洋生物中体现,陆地生物也同样受这一机制调控㊂BaP通过影响父本基因,最终影响胚胎发育㊂用BaP染毒的雄性小鼠进行体外受精后,发现在8-细胞期和囊胚期存在基因表达异常,包括调控细胞周期以及DNA修复的基因[39]㊂妊娠黄体可以分泌雌孕激素,在生殖系统中发挥重要作用,黄体的发育与血管内皮生成因子有着密切联系[40]㊂苯并芘可以使血管内皮生成因子相关基因,如血管生成素-1(an-giopoietin-1,Ang-1)㊁血管内皮细胞生长因子受体(vascular endothelial growth factor,VEGFR)㊁内皮细胞TEK酪氨酸激酶表达下调,并增加抗血管生成因子血小板反应蛋白(recombinant thrombospondin1, THBS1)的表达,还影响了对黄体血管系统建立至关重要的基因Notch1㊁DLL4㊁Jag1和Hay2的表达,破坏了黄体血管网络系统的形成,最终影响了妊娠过程中黄体的内分泌功能[41]㊂综上所述,在3种BaP发挥致毒作用的机制中(图1),BaP诱导生殖发育相关基因表达异常或提高促凋亡基因表达起主导作用,也是目前研究较为透彻的机制(图2),下面将从雄性生殖㊁雌性生殖以及胚胎发育3个角度详述BaP的毒性机制㊂2㊀BaP的雄性生殖毒性(Male reproductive toxici-ty of BaP)2.1㊀BaP对雄性激素的毒性(Toxicity of BaP to an-drogens)BaP作为内分泌干扰物主要影响睾酮水平[42],睾酮主要是由睾丸间质细胞合成分泌的,其主要成分为类固醇㊂BaP可以降低睾酮的转化率[43]和(或)睾酮的浓度[44]㊂有研究表明睾丸巨噬细胞分泌的白介素1β(interleukin-1β,IL-1β)和肿瘤坏死因子α168㊀生态毒理学报第19卷(tumor necrosis factor α,TNF α)通过抑制类固醇生成急性调节蛋白(steroidogenic acute regulatory protein,STAR)表达进一步抑制间质细胞合成睾酮[45]㊂Zheng 等[46]发现BaP 通过增加IL -1β的表达,显著抑制雄性大鼠睾酮的产生,他们还发现BaP 可以改变睾丸巨噬细胞亚群,激活ED2+睾丸巨噬细胞并促进了IL -1β的产生,最终抑制雄性大鼠睾酮合成㊂此外,3β-羟基类固醇脱氢酶(3β-hydroxysteroid dehy -drogenase,3β-HSD)与细胞色素P450胆固醇侧链裂解酶(cholesterol side -chain lyase P450scc,P450scc)在间质细胞合成睾酮中起着重要作用[47],其表达改变时会影响睾酮水平;STAR 表达的下调也可以导致睾酮合成减少[48-49]㊂雄性大鼠用BaP 灌胃90d 后,检测到BaP 下调间质细胞中的STAR ㊁3β-HSD 以及细胞色素P45017A1(cytochrome P450family 17subfamily A member 1,CYP17A1)表达,并上调P450scc 表达,进而降低大鼠睾丸间质细胞生成睾酮的能力[50]㊂Sheweita 等[51]发现BaP 降低类固醇合成酶CYP17A1和17β-羟基类固醇脱氢酶(17β-hydroxysteroid dehydrogenase,17β-HSD)蛋白表达,使大鼠血浆睾酮浓度降低㊂Banerjee 等[52]进一步验证了BaP 通过抑制类固醇生成蛋白表达,如细胞色素P450ⅡA1(cytochrome P450family Ⅱsubfamily A member 1,CYP ⅡA1)㊁STAR ㊁3β-HSD ㊁17β-HSD ,进一步降低血清睾酮水平,2021年Daoud 等[53]再一次证实了上述观点㊂Yang 等[54]发现BaP 也可以通过影响3β-HSD ㊁CYP17和17β-HSD 表达进一步扰乱雄性栉孔扇贝的激素水平㊂Booc 等[55]研究发现BaP 可降低雄性底鳉的睾酮水平,与其他动物不同的是BaP 并非通过调控类固醇相关基因表达造成这一结果,而是可能通过精原细胞包囊大小进而影响睾酮水平㊂综上所述,BaP 主要通过改变类固醇生成相关基因与酶的表达,抑制睾酮的生成,对雄性的生殖发育产生不利影响㊂epoxide(BPDE)图1㊀BaP 致毒途径机制注:AHR 表示芳香族化合物受体,ARNT 表示芳香族化合物受体核转运蛋白,HSP90表示热休克蛋白90,CYP450表示细胞色素P450,CYP17A1表示细胞色素P45017A1,STAR 表示类固醇生成急性调节蛋白,3β-HSD 表示3β-羟基类固醇脱氢酶,17β-HSD 表示17β-羟基类固醇脱氢酶,Caspase -3表示半胱氨酸蛋白酶-3,Caspase -9表示半胱氨酸蛋白酶-9,Bax 表示Bcl -2相关X 蛋白㊂Fig.1㊀Mechanism of BaP toxicity pathwayNote:AHR represents aryl hydrocarbon receptor,ARNT represents aryl hydrocarbon receptor nuclear transporter,HSP90represents heat shock protein 90,CYP450represents cytochrome P450family,CYP17A1represents cytochrome P450family 17subfamily A member 1,STAR represents steroidogenic acute regulatory protein,3β-HSD represents 3β-hydroxysteroid dehydrogenase,17β-HSD represents 17β-hydroxysteroid dehydrogenase,and Bax represents Bcl -2associated X protein.第2期王惠增等:苯并[a]芘对生殖系统的毒性作用及其机制研究进展169㊀图2㊀BaP通过基因调控引起生殖毒性注:GnRH2表示促性腺激素释放激素,GnRH3表示促性腺激素释放激素,IL-1β表示白介素1β,CYP17A1表示细胞色素P45017A1,STAR表示类固醇生成急性调节蛋白,3β-HSD表示3β-羟基类固醇脱氢酶,17β-HSD表示17β-羟基类固醇脱氢酶,CYP1A1代表细胞色素P4501A1, P450scc代表细胞色素P450胆固醇侧链裂解酶,Adcy-PKA代表上游腺苷环化酶-蛋白激酶,Caspase-3表示半胱氨酸蛋白酶-3,Caspase-9表示半胱氨酸蛋白酶-9,Bax表示Bcl-2相关X蛋白,Hsp90aB1代表90kDa热休克蛋白aB1,VTG代表卵黄蛋白原,CD34代表分化簇34, AMH代表抗缪勒管激素,CCND2代表细胞周期蛋白D2,FOXO1代表叉头框蛋白O1,HoxA10代表同源盒基因,BMP2代表骨形态发生蛋白-2,IBA1代表离子钙结合衔接分子1,SNCA代表重组人α-突触核蛋白,CYP19a代表细胞色素P450家族19亚家族a㊂Fig.2㊀BaP causes reproductive toxicity through gene regulationNote:GnRH2represents gonadotropin-releasing hormone2,GnRH3represents gonadotropin-releasing hormone3,IL-1βrepresents interleukin-1β, CYP17A1represents cytochrome P450family17subfamily A member1,STAR represents steroidogenic acute regulatory protein,3β-HSD represents3β-hydroxysteroid dehydrogenase,17β-HSD represents17β-hydroxysteroid dehydrogenase,CYP1A1represents cytochrome P450family1subfamily A member1,P450scc represents cholesterol side-chain lyase P450scc,Adcy-PKA represents adenylate cyclase-protein kinase,Bax represents Bcl-2associated X protein,Hsp90aB1represents recombinant heat shock protein90kDa alpha B1, VTG represents vitellogenin,CD34represents cluster designation34,AMH represents anti-Müllerian hormone, CCND2represents cyclin-D2,FOXO1represents forkhead box O1,HoxA10represents homeobox A10,BMP2represents bone morphogenetic protein-2,IBA1represents ionized calcium-binding adapter molecule1,SNCA representsrecombinant human alpha-synuclein,CYP19a represents cytochrome P450family19subfamily a.2.2㊀BaP对精子的毒性(Toxicity of BaP to sperm) 2.2.1㊀BaP减少精子生成(BaP reduces spermato-genesis)哺乳动物雄性生殖器官主要有睾丸㊁附睾㊁输精管等,其中睾丸的主要作用是生成精子和产生雄性激素,BaP主要通过损害睾丸进一步影响精子生成㊂BaP通过氧化应激或基因调控介导睾丸细胞凋亡,影响睾丸功能受损,减少精子数量㊂Banerjee等[52]证实BaP激活P38蛋白激酶(P38mitogen activated protein kinase,P38MAPK)通路来增加睾丸细胞内ROS,并降低细胞中的抗氧化酶活性[56],使睾丸细胞氧化应激损伤,减少精子的生成㊂Sheweita等[51]研究发现BaP通过降低睾丸组织中抗氧化酶CAT㊁SOD㊁GPX的活性,增加ROS水平,导致睾丸细胞线粒体膜破裂,进而引起睾丸组织凋亡㊂BaP还可通过AHR途径降低睾丸中CAT㊁SOD活性,升高H2O2含量,诱导睾丸细胞氧化应激,影响睾丸功能[57]㊂上述均为BaP对小鼠的生殖毒性,Tian等[58]发现BaP通过可引起雄性栉孔扇贝精巢氧化应激损伤,进一步减少精子生成㊂此外,BaP可以通过基因调控诱导睾丸细胞凋亡,提高睾丸细胞内的凋亡蛋白半胱氨酸蛋白酶-3(Caspase-3)和半胱氨酸蛋白170㊀生态毒理学报第19卷酶-9(Caspase-9)表达;促进细胞色素C转位到细胞质,启动线粒体凋亡途径,导致睾丸细胞凋亡,进一步导致精子生成减少[52,59]㊂BaP不仅通过影响睾丸功能减少精子生成,而且可以直接影响精子生成过程㊂Verhofstad等[60]的研究表明在精子发育各个阶段均可以检测到BPDE 导致的精子DNA损伤,这也是精子数量减少的原因之一㊂BaP可以导致雄鼠精子功能缺陷以及生育能力下降,并且Mohamed等[61]的实验证明了BaP的生殖毒性具有遗传性,但毒性随着子代数增加逐渐减弱㊂BaP可以减少精母细胞和次级精母细胞进入中晚期粗线期,阻止减数分裂过程的完成,导致精子生成减少[62]㊂此外,BaP诱导的氧化应激会降低精原细胞的存活率,并且通过下调基质金属蛋白酶(matrix metalloproteinase,MMP)水平以及上调促凋亡因子Caspase-3和Caspase-9表达促进精原细胞凋亡[63]㊂BaP作为广泛存在于生态系统中的环境污染物,不仅使陆地雄性动物精子生成异常,还影响水生生态系统中的雄性动物的精子生成㊂BaP可以通过基因调控扰乱雄性栉孔扇贝的精子发生相关基因:细胞周期蛋白D2(cyclin-D2,CCND2),联会复合体3㊁核呼吸因子1和水通道蛋白9,进一步减少精子生成[54]㊂斑马鱼胚胎暴露于BaP后,其睾丸中生殖细胞特异基因的启动子发生甲基化上调,进一步下调相关基因表达,最终抑制精子生成,影响雄性斑马鱼的生殖能力[64]㊂2.2.2㊀BaP降低精子活力(BaP reduces sperm motility)BaP可以损害睾丸和附睾的内分泌功能,从而导致储存的精子活力下降[65-67]㊂睾丸的质量和大小与精子的数量和活力成正比[68],雄性小鼠用BaP连续灌胃60d,检测到小鼠的睾丸质量明显降低,精子的活力也随之降低[69]㊂小鼠暴露于BaP后,其睾丸支持细胞和间质细胞均凋亡,进而影响精子发生过程,最终导致精子活力减弱[69]㊂畸形精子的活力及存活率显著低于正常精子,BaP暴露会导致精子形态异常,畸形精子大幅增加,主要异常表现为无尾㊁双头㊁中段弯曲[57]㊂Xu等[59]验证了BaP可导致精子活动力降低,精子头㊁尾部畸形率以及总畸形率均显著升高㊂最新研究表明BaP改变睾丸激素水平引起雄性交配强度减弱,降低精子质量,引起畸形精子增多[70-71]㊂有研究表明精子短端粒可能是导致男性不育的原因之一[72],Ling等[73]研究发现BaP可以使精子端粒变短,且与剂量成反比㊂3㊀BaP的雌性生殖毒性(Female reproductive toxicity of BaP)3.1㊀BaP对雌性激素的毒性(Toxicity of BaP to es-trogen)BaP作为一种常见的环境污染物,是海洋环境污染原因之一,影响水生动物的繁殖㊂雌孕激素对雌性发育有不可或缺的作用,而BaP作为内分泌干扰物可以降低水生动物血浆中的孕酮㊁雌激素和催乳素浓度[74]㊂为进一步探究其发生机制,Tian等[58]用不同浓度的BaP处理雌性栉孔扇贝,发现BaP可以导致类固醇合成相关酶(3β-HSD㊁CYP17㊁17β-HSD)表达下降,并呈剂量依赖性;高浓度的BaP还可抑制AHR㊁ARNT㊁CYP1A1以及17β-雌二醇-雌激素受体转录,2种机制相辅相成,共同抑制雌孕激素的生成㊂BaP通过干扰激素膜受体降低三疣梭子蟹的雌二醇(estradiol,E2)浓度[75]㊂斑马鱼胚胎暴露于BaP会导致成年雌鱼卵巢中E2水平下降,其机制为雌鱼脑中促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)基因中的GnRH3的甲基化水平显著升高,并下调GnRH3mRNA表达,从而影响E2的产生[76]㊂与斑马鱼报道相反的是BaP可促进雌性海马GnRH2和GnRH3mRNA的表达,并导致血浆中E2水平显著下降[77]㊂2种相反结果可能与BaP的浓度㊁作用时间以及实验对象不同有关㊂Yang等[78]发现BaP抑制雌性栉孔扇贝的上游腺苷环化酶-蛋白激酶(adenylate cyclase-protein ki-nase,Adcy-PKA)信号通路,下调促性腺激素受体转录水平,如促卵泡激素受体(follicle-stimulating hor-mone receptor,FSHR)和黄体生成素/绒毛膜促性腺激素受体(luteinizing hormone/choriogonadotropin re-ceptor,LHCGR),导致类固醇生成酶(3β-HSD㊁CYP17㊁17β-HSD)表达减少,最终引起抗雌激素效应㊂Kennedy和Smyth[79]发现雌鲑鱼体内E2的减少并非是通过常规的BaP作用于类固醇机制,而是通过其他内分泌干扰机制来对抗雌激素的方式改变了血浆E2的浓度,这种机制有待进一步研究㊂综上所述,BaP主要通过基因表达调控这一途径降低雌性体内E2和孕酮水平,进而影响雌性的生殖发育㊂3.2㊀BaP对卵巢的毒性(Toxicity of BaP to ovary)卵巢是雌性生殖发育中最重要的生殖器官,具有排卵和内分泌功能,对维持雌性激素水平至关重要,暴露于BaP会扰乱卵巢的结构与功能,进一步第2期王惠增等:苯并[a]芘对生殖系统的毒性作用及其机制研究进展171㊀影响生育㊁妊娠㊂高剂量的BaP可以导致卵巢细胞退化并出现管状结构,而这些组织学变化属于癌前病变[80]㊂Rahmani等[81]发现BaP通过氧化应激导致卵巢表面上皮内陷㊁细胞堆积㊁管状结构形成,卵巢间质出现间质水肿㊁出血等病理学改变,并且BaP 诱导卵巢中Caspase-3表达升高,影响卵巢的生理功能,与睾丸相比,BaP对卵巢的危害更严重,这是因为在BaP处理后,胎儿卵巢中促细胞凋亡蛋白Bcl-2相关X蛋白(Bcl-2associated X protein,Bax)表达增加,并激活下游Caspase-3和Caspase-9,导致卵巢细胞凋亡[82]㊂卵黄蛋白原(vitellogenin,VTG)和CCND2是雌激素介导的卵巢发育相关基因[83],BaP可以下调VTG和CCND2表达,造成雌性栉孔扇贝卵巢受损,组织学检查发现,BaP可引起卵巢发育延迟和卵母细胞退化,并且卵巢的病变情况随着染毒时间和染毒剂量的增加而严重[78]㊂研究发现BaP可以抑制脂联素受体1(adiponectin receptor protein1,AdipoR1)和脂联素受体2(adiponectin receptor protein2,AdipoR2)表达,进而影响卵巢功能[84]㊂最新研究表明,BaP及其代谢产物BPDE可抑制妊娠小鼠卵巢中腺嘌呤核苷酸转运体1(adenine nucleotide translocator1,ANT1)的表达,进一步研究发现ANT1的过表达可以修复BPDE引起的有丝分裂缺陷,恢复卵巢功能[85]㊂3.3㊀BaP对雌性生殖细胞的毒性(Toxicity of BaP to female germ cells)3.3.1㊀BaP影响卵泡发生和发育(BaP affects folli-cular genesis and development)卵泡发育是女性正常的生理过程,卵泡的发育情况直接关系到后代繁殖㊂卵泡作为卵巢的功能单位,支持卵母细胞的发育和成熟[86]㊂卵泡的生长发育过程相当复杂,原始卵泡经历初级卵泡㊁窦前㊁窦卵泡才能发育为成熟卵泡[87-88]㊂有报道称BaP作为卵毒物质,可以破坏原始卵泡[89],或者使原始卵泡迅速枯竭[90],BaP还可以通过香烟烟雾进入卵泡液中,对卵泡发育产生不利影响[91]㊂Sobinoff等[24]研究了BaP卵毒性的机制,连续用BaP处理雌性小鼠7d会导致卵巢中的原始卵泡显著减少,卵泡闭锁,其具体机制为BaP通过干扰AHR发育信号破坏卵泡形成㊂有报道称BaP不仅可以减少或耗尽原始卵泡和初级卵泡的数量[89-90],还可以抑制卵泡生长发育,即BaP处理过的卵泡均发育不到窦前阶段[92]㊂Sadeu和Foster[93]将小鼠卵泡暴露于不同浓度的BaP,发现卵泡存活率均降低,其中高浓度的BaP会抑制窦卵泡发育,使卵泡停滞于窦前卵泡阶段,窦卵泡比例显著减少㊂抗缪勒管激素(anti-Müllerian hormone,AMH)浓度增加与卵泡发育停滞有关[94-95]㊂有研究表明BaP可以通过减少AMH生成,促进卵泡募集到卵泡池中,最终加快卵泡枯竭的速度[96]㊂Sadeu和Foster[93]进一步探索了BaP诱导卵泡发育异常的关键分子途径,发现BaP暴露通过激活窦前㊁窦卵泡和成熟卵泡中AHR信号通路,进一步促进促凋亡因子Bax激活,此外,BaP暴露还会导致90kDa热休克蛋白aB1(recombinant heat shock protein90kDa alpha B1,Hsp90aB1)基因表达上调,导致卵泡生长延迟和存活率下降㊂卵泡生长和卵泡发育在雌性哺乳动物生殖中有着重要地位,BaP不但可以通过基因表达调控导致卵泡生长发育异常,还可能通过氧化应激影响卵泡发育㊂3.3.2㊀BaP影响卵母细胞功能(BaP affects oocyte function)BaP可使卵母细胞线粒体内ROS水平升高,导致精-卵结合和融合障碍,影响动物的繁殖[24,36]㊂BaP可导致卵母细胞和卵丘细胞DNA断裂,细胞功能障碍,影响卵母细胞进一步发育,精卵融合失败[34],这也是BaP生殖毒性机制之一㊂卵母细胞的减数分裂在卵母细胞成熟与成功受精中起着重要作用[97],BaP诱导卵母细胞减数分裂异常,卵母细胞功能障碍,不利于动物繁殖㊂BaP可以阻滞猪卵母细胞减数分裂,使部分卵母细胞停滞在MⅡ期,进一步检测发现BaP通过降低乙酰化α-微管蛋白,导致微管不稳定,损害纺锤体组装,从而干扰卵母细胞减数分裂过程[36]㊂Sui等[98]通过将雌鼠暴露于BaP检测其对子代的影响,验证了BaP对卵母细胞的遗传毒性:生发泡破裂(germinal vesicle breakdown, GVBD)是卵母细胞成熟的关键事件,母体暴露BaP 会降低子代GVBD率[99];并且BaP会扰乱子代卵母细胞的纺锤体组装和染色体配对,使卵母细胞减数分裂停滞;最后,雌鼠暴露于BaP可导致子代卵母细胞基因组高甲基化,损害卵母细胞的发育能力㊂综上所述,母系BaP暴露损害了子代卵母细胞的进一步发育,这与上文Miao等[36]研究结果相一致㊂4㊀BaP对胎儿或胚胎的生殖毒性(Reproductive toxicity of BaP to the fetus or embryo)4.1㊀BaP的胚胎发育毒性(Embryonic developmen-tal toxicity of BaP)早有研究表明,吸烟损害身体健康,还对孕妇以。

第 63 卷第 2 期2024 年 3 月Vol.63 No.2Mar.2024中山大学学报（自然科学版）（中英文）ACTA SCIENTIARUM NATURALIUM UNIVERSITATIS SUNYATSENI红树植物桐花树内生真菌Talaromyces amestolkiae 30的次生代谢产物*刘洪亮1，赵飞1，唐凤婷1，李锦俊1，张轩1，杜志云1，黄华容1，佘志刚21. 广东工业大学生物医药学院，广东广州 5100062. 中山大学化学学院，广东广州 510006摘要：对红树植物桐花树内生真菌Talaromyces amestolkiae 30的次级代谢产物进行了研究。

采用大米固体发酵培养，色谱分离技术纯化单体，ESIMS和NMR等波谱数据分析，鉴定了12个异香豆素单体化合物：aspergillumarin A（1）、aspergillumarin B（2）、 5，6-dihydroxy-3-（4-hydroxypentyl）-isochroman-1-one（3）、 mucoriso‐coumarin A（4）、 peniisocoumarin H（5）、 peniisocoumarin E（6）、 dichlorodiaportin（7）、 mucorisocoumarin C（8）、 peniiso‐coumarin G（9）、 talumarin A（10）、 5，6，8-trihydroxy-4-（1'-hydroxyethyl）-isocoumarin（11）和sescandelin（12），其中化合物4、6、7首次从篮状属真菌中分离得到。

二倍稀释法抑菌活性测试显示，化合物4、6、7有抑制金黄色葡萄球菌作用；MTT法测试细胞毒活性，表明化合物7对前列腺癌PC-3细胞和VCaP细胞有细胞毒活性。

关键词：红树林真菌；篮状菌；次级代谢产物；异香豆素类中图分类号：O629.9 文献标志码：A 文章编号：2097 - 0137（2024）02 - 0160 - 08The metabolites from mangrove endophytic fungus Talaromyces amestolkiae30LIU Hongliang1， ZHAO Fei1， TANG Fengting1， LI Jinjun1，ZHANG Xuan1， DU Zhiyun1， HUANG Huarong1， SHE Zhigang21. School of Biomedicine， Guangdong University of Technology， Guangzhou 510006， China2. School of Chemistry， Sun Yat-sen University， Guangzhou 510006， ChinaAbstract：The metabolites of the endophytic fungus Talaromyces amestolkiae30 from the mangrove plant Aegiceras corniculatum (L.) Blanco were investigated. The fungus was cultured in rice medium, the monomeric compounds were isolated and purified by the chromatographic technique, and the structures of the compounds were identified by analysis of spectroscopy such as ESIMS and NMR.Twelve known analogues of isocoumarins (1-12) were isolated and identified as aspergillumarin A (1), aspergillumarin B (2), 5,6-dihydroxy-3-(4-hydroxypentyl)-isochroman-1-one (3), mucorisocoumarin A(4), peniisocoumarin H (5), peniisocoumarin E (6), dichlorodiaportin (7), mucorisocoumarin C (8), peni‐isocoumarin G (9), talumarin A (10), 5,6,8-trihydroxy-4-(1'-hydroxyethyl)-isocoumarin (11) and sescan‐delin (12). Among them, compounds 4, 6 and 7 were obtained from the genus Talaromyces for the first time. The antibacterial activities of these compounds were tested in vitro using the twofold dilution method. Compounds 4, 6, and 7 showed inhibitory activity against Staphylococcus aureus. The cytotoxic activity was tested by the MTT assay. Compound 7 showed cytotoxicity against prostate cancer PC-3DOI：10.13471/ki.acta.snus.2023E048*收稿日期：2023 − 10 − 14 录用日期：2023 − 11 − 30 网络首发日期：2024 − 01 − 08基金项目：广东省海洋经济发展（海洋六大产业）专项资金（粤自然资合［2021］48号）作者简介：刘洪亮（1996年生），男；研究方向：制药工程；E-mail：*********************通信作者：黄华容（1978年生），女；研究方向：天然药物化学；E-mail：****************.cn第 2 期刘洪亮，等：红树植物桐花树内生真菌Talaromyces amestolkiae 30的次生代谢产物cells and VCaP cells.Key words ： mangrove fungus ； Talaromyces amestolkiae ； secondary metabolites ； isocoumarin 异香豆素是一类重要的天然产物，广泛存在于自然界中，其种类繁多（Saddiqa et al.，2017； Reveglia et al.，2020；Shabir et al.，2021；Aierken et al.，2023）。

Clinical Policy: Ado-Trastuzumab Emtansine (Kadcyla) Reference Number: CP.PHAR.229Effective Date: 06.01.16Last Review Date: 05.23Line of Business: Commercial, HIM, Medicaid Coding ImplicationsRevision LogSee Important Reminder at the end of this policy for important regulatory and legal information.DescriptionAdo-trastuzumab emtansine (Kadcyla®) is a human epidermal growth factor receptor 2 protein (HER2)-targeted antibody and microtubule inhibitor conjugate.FDA Approved Indication(s)Kadcyla is indicated as a single agent for the:•Adjuvant treatment of patients with HER2-positive early breast cancer who have residual invasive disease after neoadjuvant taxane and trastuzumab-based treatment.•Treatment of patients with HER2-positive, metastatic breast cancer who previously received trastuzumab and a taxane, separately or in combination. Patients should have either:o Received prior therapy for metastatic disease, oro Developed disease recurrence during or within six months of completing adjuvant therapy.Policy/CriteriaProvider must submit documentation (such as office chart notes, lab results or other clinical information) supporting that member has met all approval criteria.It is the policy of health plans affiliated with Centene Corporation® that Kadycla is medically necessary when the following criteria are met:I.Initial Approval CriteriaA.Breast Cancer (must meet all):1.Diagnosis of HER2-positive breast cancer;2.Prescribed by or in consultation with an oncologist;3.Age ≥ 18 years;4.Prescribed as a single agent;5.Documentation of prior use of trastuzumab-based therapy and a taxane;6.Request meets one of the following (a, b, or c):*a.As adjuvant: Dose does not exceed 3.6 mg/kg every 21 days for a maximum of 14doses;b.For metastatic: Dose does not exceed 3.6 mg/kg every 21 days;c.Dose is supported by practice guidelines or peer-reviewed literature for therelevant off-label use (prescriber must submit supporting evidence).*Prescribed regimen must be FDA-approved or recommended by NCCNApproval duration:Medicaid/HIM – 6 monthsCLINICAL P OLICYAdo-Trastuzumab EmtansineCommercial – 6 months or to the member’s renewal date, whichever is longerB.Additional NCCN Recommended Uses (off-label) (must meet all):1.Diagnosis of one of the following (a or b):a.Recurrent, advanced, or metastatic HER2-positive non-small cell lung cancer(NSCLC);b.Recurrent HER2-positive salivary gland tumor;2.Prescribed by or in consultation with an oncologist;3.Age ≥ 18 years;4.Prescribed as a single agent;5.Request meets one of the following (a or b):*a.Dose does not exceed 3.6 mg/kg every 21 days;b.Dose is supported by practice guidelines or peer-reviewed literature for therelevant off-label use (prescriber must submit supporting evidence).*Prescribed regimen must be FDA-approved or recommended by NCCNApproval duration:Medicaid/HIM – 6 monthsCommercial – 6 months or to the member’s renewal date, whichever is longerC.Other diagnoses/indications (must meet 1 or 2):1.If this drug has recently (within the last 6 months) undergone a label change (e.g.,newly approved indication, age expansion, new dosing regimen) that is not yetreflected in this policy, refer to one of the following policies (a or b):a.For drugs on the formulary (commercial, health insurance marketplace) or PDL(Medicaid), the no coverage criteria policy for the relevant line of business:CP.CPA.190 for commercial, HIM.PA.33 for health insurance marketplace, andCP.PMN.255 for Medicaid; orb.For drugs NOT on the formulary (commercial, health insurance marketplace) orPDL (Medicaid), the non-formulary policy for the relevant line of business:CP.CPA.190 for commercial, HIM.PA.103 for health insurance marketplace, andCP.PMN.16 for Medicaid; or2.If the requested use (e.g., diagnosis, age, dosing regimen) is NOT specifically listedunder section III (Diagnoses/Indications for which coverage is NOT authorized) ANDcriterion 1 above does not apply, refer to the off-label use policy for the relevant lineof business: CP.CPA.09 for commercial, HIM.PA.154 for health insurancemarketplace, and CP.PMN.53 for Medicaid.II.Continued TherapyA.All Indications in Section I (must meet all):1.Currently receiving medication via Centene benefit, or documentation supports thatmember is currently receiving Kadcyla for a covered indication and has received thismedication for at least 30 days;2.Member is responding positively to therapy;3.If request is for a dose increase, request meets one of the following (a, b, or c):*a.As adjuvant therapy for breast cancer: New dose does not exceed 3.6 mg/kg every21 days for a maximum of 14 doses;CLINICAL P OLICYAdo-Trastuzumab Emtansineb.For all other indications: New dose does not exceed 3.6 mg/kg every 21 days;c.New dose is supported by practice guidelines or peer-reviewed literature for therelevant off-label use (prescriber must submit supporting evidence).*Prescribed regimen must be FDA-approved or recommended by NCCNApproval duration:Medicaid/HIM – 12 monthsCommercial – 6 months or to the member’s renewal date, whichever is longerB.Other diagnoses/indications (must meet 1 or 2):1.If this drug has recently (within the last 6 months) undergone a label change (e.g.,newly approved indication, age expansion, new dosing regimen) that is not yetreflected in this policy, refer to one of the following policies (a or b):a.For drugs on the formulary (commercial, health insurance marketplace) or PDL(Medicaid), the no coverage criteria policy for the relevant line of business:CP.CPA.190 for commercial, HIM.PA.33 for health insurance marketplace, andCP.PMN.255 for Medicaid; orb.For drugs NOT on the formulary (commercial, health insurance marketplace) orPDL (Medicaid), the non-formulary policy for the relevant line of business:CP.CPA.190 for commercial, HIM.PA.103 for health insurance marketplace, andCP.PMN.16 for Medicaid; or2.If the requested use (e.g., diagnosis, age, dosing regimen) is NOT specifically listedunder section III (Diagnoses/Indications for which coverage is NOT authorized) ANDcriterion 1 above does not apply, refer to the off-label use policy for the relevant lineof business: CP.CPA.09 for commercial, HIM.PA.154 for health insurancemarketplace, and CP.PMN.53 for Medicaid.III.D iagnoses/Indications for which coverage is NOT authorized:A.Non-FDA approved indications, which are not addressed in this policy, unless there issufficient documentation of efficacy and safety according to the off label use policy –CP.CPA.09 for commercial, HIM.PA.154 for health insurance marketplace, andCP.PMN.53 for Medicaid, or evidence of coverage documents.IV.Appendices/General InformationAppendix A: Abbreviation/Acronym KeyFDA: Food and Drug AdministrationHER2: human epidermal growth factor receptor 2 proteinNSCLC: non-small cell lung cancerAppendix B: Therapeutic AlternativesNot applicableAppendix C: Contraindications/Boxed Warnings•Contraindication(s): none reported•Boxed warning(s): hepatotoxicity, cardiac toxicity, and embryo-fetal toxicityC LINICAL P OLICYAdo-Trastuzumab EmtansineV. Dosage and AdministrationMaximum Dose Breast cancer Adjuvant therapy for early breast cancer with residual disease3.6 mg/kg IV Q3WK (21-day cycle) for a total of 14 cycles unless there is disease recurrence or unmanageable toxicity.Metastatic breast cancer3.6 mg/kg IV Q3WK (21-day cycle) until disease progression or unmanageable toxicity.3.6 mg/kgVI. Product AvailabilitySingle-use vial: 100 mg, 160 mgVII. References1. Kadcyla Prescribing Information. South San Francisco, CA: Genentech, Inc.; February 2022. Available at: https:///download/pdf/kadcyla_prescribing.pdf. Accessed January 4, 2023.2. Ado-trastuzumab emtansine. In: National Comprehensive Cancer Network Drugs andBiologics Compendium. Available at: /professionals/drug_compendium. Accessed February 7, 2023.3. Minckwitz GV, Huang CS, Mano MS, et al. Trastuzumab emtansine for residual invasive HER2-positive breast c ancer. N Engl J Med 2019;380:617-28.4. National Comprehensive Cancer Network Guidelines. Breast Cancer Version 2.2023. Available at https:///professionals/physician_gls/pdf/breast.pdf. Accessed February 7, 2023.Coding ImplicationsCodes referenced in this clinical policy are for informational purposes only. Inclusion orexclusion of any codes does not guarantee coverage. Providers should reference the most up-to-date sources of professional coding guidance prior to the submission of claims for reimbursement of covered services. J9354 Injection, ado-trastuzumab emtansine, 1 mgDate P&TApproval Date2Q 2019 annual review: expanded COC to all covered indications from just breast cancer; references reviewed and updated.02.05.19 05.19 Criteria added for new FDA indication: adjuvant therapy in early breast cancer with residual disease; references reviewed and updated.06.11.19 08.19C LINICAL P OLICYAdo-Trastuzumab EmtansineDateP&T Approval Date2Q 2020 annual review: no significant changes; revised HIM-medical benefit to HIM line of business; references reviewed and updated. 02.18.20 05.20 2Q 2021 annual review: combined NSCLC and new off-label salivary gland tumor indications supported by NCCN into one off-label section under I.B.; references for HIM line of business off-label use revised from HIM.PHAR.21 to HIM.PA.154; references reviewed and updated.02.05.2105.212Q 2022 annual review: added criterion for single-agent therapy for off-label indications of NSCLC and salivary gland tumor per NCCN; references reviewed and updated.02.15.22 05.22 Template changes applied to other diagnoses/indications.10.07.22 2Q 2023 annual review: no significant changes; clarified for NSCLC that disease is recurrent, advanced, or metastatic per NCCN; references reviewed and updated.01.04.2305.23Important ReminderThis clinical policy has been developed by appropriately experienced and licensed health care professionals based on a review and consideration of currently available generally accepted standards of medical practice; peer-reviewed medical literature; government agency/program approval status; evidence-based guidelines and positions of leading national health professional organizations; views of physicians practicing in relevant clinical areas affected by this clinical policy; and other available clinical information. The Health Plan makes no representations and accepts no liability with respect to the content of any external information used or relied upon in developing this clinical policy. This clinical policy is consistent with standards of medicalpractice current at the time that this clinical policy was approved. “Health Plan” means a health plan that has adopted this clinical policy and that is operated or administered, in whole or in part, by Centene Management Company, LLC, or any of such health plan’s affiliates, as applicable.The purpose of this clinical policy is to provide a guide to medical necessity, which is a component of the guidelines used to assist in making coverage decisions and administering benefits. It does not constitute a contract or guarantee regarding payment or results. Coverage decisions and the administration of benefits are subject to all terms, conditions, exclusions and limitations of the coverage documents (e.g., evidence of coverage, certificate of coverage, policy, contract of insurance, etc.), as well as to state and federal requirements and applicable Health Plan-level administrative policies and procedures.This clinical policy is effective as of the date determined by the Health Plan. The date of posting may not be the effective date of this clinical policy. This clinical policy may be subject to applicable legal and regulatory requirements relating to provider notification. If there is a discrepancy between the effective date of this clinical policy and any applicable legal orregulatory requirement, the requirements of law and regulation shall govern. The Health Plan retains the right to change, amend or withdraw this clinical policy, and additional clinical policies may be developed and adopted as needed, at any time.CLINICAL P OLICYAdo-Trastuzumab EmtansineThis clinical policy does not constitute medical advice, medical treatment or medical care. It is not intended to dictate to providers how to practice medicine. Providers are expected to exercise professional medical judgment in providing the most appropriate care, and are solely responsible for the medical advice and treatment of members. This clinical policy is not intended to recommend treatment for members. Members should consult with their treating physician in connection with diagnosis and treatment decisions.Providers referred to in this clinical policy are independent contractors who exercise independent judgment and over whom the Health Plan has no control or right of control. Providers are not agents or employees of the Health Plan.This clinical policy is the property of the Health Plan. Unauthorized copying, use, and distribution of this clinical policy or any information contained herein are strictly prohibited. Providers, members and their representatives are bound to the terms and conditions expressed herein through the terms of their contracts. Where no such contract exists, providers, members and their representatives agree to be bound by such terms and conditions by providing services to members and/or submitting claims for payment for such services.Note: For Medicaid members, when state Medicaid coverage provisions conflict with the coverage provisions in this clinical policy, state Medicaid coverage provisions take precedence. Please refer to the state Medicaid manual for any coverage provisions pertaining to this clinical policy.©2016 Centene Corporation. All rights reserved. All materials are exclusively owned by Centene Corporation and are protected by United States copyright law and international copyright law. No part of this publication may be reproduced, copied, modified, distributed, displayed, stored in a retrieval system, transmitted in any form or by any means, or otherwise published without the prior written permission of Centene Corporation. You may not alter or remove any trademark, copyright or other notice contained herein. Centene® and Centene Corporation® are registered trademarks exclusively owned by Centene Corporation.。

生物学英语复试题及答案一、选择题（每题2分，共20分）1. Which of the following is not a characteristic of living organisms?A. ReproductionB. GrowthC. Response to stimuliD. Inanimate objects2. What is the basic unit of life?A. CellB. OrganC. TissueD. Organ system3. What is the process by which organisms convert sunlight into chemical energy?A. RespirationB. PhotosynthesisC. FermentationD. Cellular respiration4. Which of the following is not a type of symbiotic relationship?A. MutualismB. CommensalismC. ParasitismD. Competition5. What is the term for the study of the structure of organisms?A. AnatomyB. PhysiologyC. EcologyD. Taxonomy6. What is the primary function of the mitochondria in a cell?A. Protein synthesisB. DNA replicationC. Energy productionD. Cell wall synthesis7. What is the term for the process by which new speciesevolve from existing ones?A. AdaptationB. Natural selectionC. SpeciationD. Genetic drift8. Which of the following is not a type of genetic mutation?A. DeletionB. InsertionC. DuplicationD. Mitosis9. What is the term for the study of the diversity of life on Earth?A. BiodiversityB. BiotechnologyC. BioinformaticsD. Biogeography10. What is the process by which organisms obtain nutrients from their environment?A. IngestionB. AssimilationC. DigestionD. Absorption二、填空题（每题2分，共20分）1. The scientific method involves making observations, forming hypotheses, conducting experiments, and then drawing ________.2. The genetic material in cells is composed of molecules called ________.3. The process by which organisms produce offspring that are similar to themselves is known as ________.4. In an ecosystem, the transfer of energy from one trophic level to the next is called ________.5. The study of the interactions between organisms and their environment is known as ________.6. The smallest unit of matter that retains the properties of an element is called a(n) ________.7. The process by which plants absorb water and nutrients from the soil is called ________.8. The study of the classification of organisms based ontheir evolutionary relationships is known as ________.9. The process by which organisms break down complex organic molecules into simpler ones is called ________.10. The study of the nervous system, including the brain,spinal cord, and nerves, is known as ________.三、简答题（每题10分，共40分）1. Explain the difference between asexual and sexual reproduction.2. Describe the role of DNA in the inheritance of traits.3. Discuss the importance of biodiversity for ecosystems.4. Explain how natural selection contributes to the evolution of species.四、论述题（20分）Discuss the impact of genetic engineering on modern agriculture and its potential ethical concerns.答案：一、选择题1. D2. A3. B4. D5. A6. C7. C8. D9. A10. A二、填空题1. Conclusions2. DNA3. Reproduction4. Energy flow5. Ecology6. Atom7. Absorption8. Phylogenetics9. Catabolism10. Neuroscience三、简答题1. Asexual reproduction involves a single organism producing offspring that are genetically identical to itself, while sexual reproduction involves the combination of genetic material from two organisms to produce offspring with a mix of traits from both parents.2. DNA carries the genetic information that determines the traits of an organism. It is passed down from parents to offspring during reproduction, allowing for the inheritance of specific characteristics.3. Biodiversity is crucial for ecosystems as it contributes to their stability, resilience, and ability to adapt to changes. It also supports a wide range of ecosystem services that are essential for human well-being.4. Natural selection is the process by which organisms that are better adapted to their environment are more likely to survive and reproduce, passing on their advantageous traits to their offspring. Over time, this leads to the evolution of species as traits that enhance survival and reproduction become more common.四、论述题Genetic engineering has revolutionized modern agriculture byenabling the development of crops with desirable traits such as disease resistance, pest resistance, and higher yields. However, it also raises ethical concerns regarding the potential impact on biodiversity, the possibility of creating "superweeds" or "superbugs," and the long-term health effects of genetically modified organisms on humans and the environment. It。

第 33 卷 第 5 期Vol.33，No.5106-1142024 年 5 月草业学报ACTA PRATACULTURAE SINICA 邹晓璐， 张文静， 吕红， 等. 醉鱼草内生细菌 ZJ1的生物学特性及防病促生效果. 草业学报， 2024， 33（5）： 106−114.ZOU Xiao -lu ， ZHANG Wen -jing ， LYU Hong ， et al . Biological characteristics and plant growth -promoting and biocontrol properties of endophytic bacterium ZJ1 from Buddleja lindleyana . Acta Prataculturae Sinica ， 2024， 33（5）： 106−114.醉鱼草内生细菌 ZJ1的生物学特性及防病促生效果邹晓璐，张文静，吕红，秦楠，赵晓军，殷辉，任璐*（山西农业大学植物保护学院，山西 太原 030031）摘要：为推动生物防治技术在实践中的应用，本研究采用稀释涂布平板法对菌株ZJ1的致死温度和酸碱度进行测定，结果表明，菌株ZJ1的致死温度为96 ℃、可存活20 min ；致死pH 为≤4或≥14。

而生物膜能够吸附土壤中的金属离子改善植物生长环境，帮助生防菌株更好地定殖在植物表面，发挥生防功能。

采用菌膜检测法观察几种常见金属离子对菌株ZJ1生物膜形成的影响，结果表明，0.3%的Zn 2+、Mn 2+、Cu 2+ 和 Fe 2+会抑制菌株ZJ1产生生物膜 ，但同浓度的Mg 2+、K +均可促进菌株ZJ1生物膜的产生；进一步采用96微孔板定量法对菌株ZJ1生物膜形成能力进行了测定，结果表明，菌株ZJ1为强生物膜形成株。

室内盆栽试验结果表明，菌株ZJ1发酵液10×稀释液对番茄早疫病的保护效果最高，达到95.54%，菌株ZJ1发酵液200×稀释液对番茄灰霉病的治疗效果最高达，到91.99%。

2023-2024学年上海市静安区高三上学期期末教学质量调研考试英语试题Directions: After reading the passage below, fill in the blanks to make the passages coherent and grammatically correct. For the blanks with a given word, fill in each blank with the proper form of the given word; for the other blanks, use one word that best fits each blank.Japan’s robot revolution in senior careJapan’s artificial intelligence expertise is transforming the elder care industry, with 1 (specialize) robotic care accomplishing more than just taking pressure off the critical shortage of caregivers. Senior care facilities across Japan are testing out such new robots 2 deliver a collection of social and physical health care and the government-backed initiative has been met with positive reviews by elderly residents.The rapidly graying population 3 (eye) by the government as a potential market for medical technology now. Disappointing government predictions show that by 2025, Japan's first baby boomers will have turned 75 and about 7 million people are likely to suffer from some form of dementia (痴呆). The nation won't be able to avoid a dementia crisis 4 an additional 380,000 senior care workers.The long-standing shortage of professional care workers has encouraged the Japanese government 5 (simplify) procedures for foreign caregivers to be trained and certified. The current Technical Intern Training Program between Vietnam, the Philippines, and Indonesia, under 6 Economic Partnership Agreement, was extended to include nursing care as well as agriculture, fishery, and construction sectors.7 the government made efforts to increase the numbers of senior care workers, the target number of foreign graduates has still fallen flat, with the national caregiver examination proving a major obstacle to pass. The success rate for foreign students was a merely 106 students last year, 8 has slightly improved to 216 students this year. Another depressing reality is that 19 to 38 percent of foreign nurses who pass the exam opt to leave the industry and return home, 9 (cite) tough work conditions and long hours. Given the challenges, this is 10 the government believes care robots will be able to step in.Directions: Complete the following passage by using the words in the box. Each word can only be used once. Note that there is one word more than you need.A. smoothingB. remainC. switchedD. likelihoodE. impactF. tipG. broadly H. headed I. booming J. positioning K. reliablySea-level rise predictionsA team of University of Idaho scientists is studying a fast-moving glacier in Alaska in hopes of developing better predictions on how quickly global sea levels will rise.Tim Bartholomaus, a professor in the Department of Geography and Geological Sciences, spent several weeks on Turner Glacier in Alaska’s southeastern 11 near Disenchantment Bay. The glacier is unique because, unlike other glaciers, it rises greatly every five to eight years.A surging glacier is defined, 12 , as one that starts flowing at least 10 times faster than normal. But the how and why of that glacial movement is poorly understood, although recent research suggests that global climate change increases the 13 of glacial surging.During Turner’s surges, the mass of ice and rock will increase its speed from roughly 3 feet a day to 65 feet per day.All of that is important because glaciers falling into the ocean are a major contributor to sea level rise, and current clima te change models don’t 14 account for these movements. For example, Greenland’s glaciers are one of the leading contributors to global sea-level rise. Since the early 2000s, Greenland 15 from not having any effect on world sea levels, to increasing sea level by about 1 millimeter per year. Half of that yearly increase is due to warmer average temperatures, which leads to more ice melting. The other half, however, is because glaciers in Greenland are, as a whole, moving faster and running into the ocean more frequently.Glacial movement has something to do with water running underneath the glacier. Glaciers are full of holes, and water runs through those holes. When the water pressure is high underneath a glacier, it starts to move, partly because it’s li fting the mass of ice and rock off the ground and partly because it’s 16 the underside of the glacier.But how exactly does that water move through the glacier, and how does the movement 17 the glacier’s speed? Those are the questions the scientists ho pe to answer.Bartholomaus, some graduate students and researchers from Boise State University, 18 onto the ice in August. They set up a base camp at the toe of the glacier and spent their days flying in on helicopters. They placed roughly 30 instruments, burying them deeply into the glacier and 19 them on rock outcroppings (露岩) alongside the glacier. This summer the team will return to get the instruments and replace batteries. Those instruments will 20 on and around the glacier until the glacier surge stops, providing researchers with before and after data.Investors probably expect that following the suggestions of stock analysts would make them better off than doing the exact opposite. _________, recent research by Nicola Gennaioli and his colleagues shows that the best way to gain excess return s would be to invest in the shares least favored by analysts. They compute that, during the last 35 years, investing in the 10 percent of U. S. stocks analysts were most _________ about would have yielded on average 3 percent a year._________, investing in the 10 percent of stocks analysts were most pessimistic about would have yielded a surprising 15 percent a year.Gennaioli and colleagues shed light on this _________ with the help of cognitive sciences and, in particular, using Kahneman and Tversky's concept of representativeness. Decision makers, according to this view, _________ the representative features of a group or a phenomenon. These are defined as the features that occur more frequently in that group than in a baseline reference group.After observing strong earnings growth—the explanation goes—analysts think that the firm may be the next Google. “Googles” are in fact more frequent among firms experiencing strong growth, which makes them _________. The problem is that “Googles” are very _________ in absolute terms. As a result, expectations become too optimistic, and future performance_________. A model of stock prices in which investor beliefs follow this logic can account both qualitatively and quantitatively for the beliefs of analysts and the dynamics (动态变化) of stock returns.In related work, the authors also show that the same model can _________ booms and busts in the volume of credit and interest rate spreads.These works are part of a research project aimed at taking insights from cognitive sciences and at__________them into economic models. Kahneman and Tversky's concept of “representativeness” lies at the heart of this effort. “In a classical example, we __________ to think of Irishmen as redheads because red hair is much more frequent among Irishmen than among the rest of the world,” Prof. Gennaioli says. “However, only 10 percent of Irishmen are redheads. In our work, we develop models of belief formation that show this logic and study the __________ of this important psychological force in different fields.”Representativeness helps describe __________ and behavior in different fields, not only in financial markets. One such field is the formation of stereotypes about social groups. In a recent experimental paper, Gennaioli and colleagues show that representativeness can explain self-confidence, and in particular the __________ of women to compete in traditionally male subjects, such as mathematics.A slight prevalence of __________ male math ability in the data is enough to make math ability un-representative for women, driving their under confidence in this particular subject.21.A．Consequently B．Furthermore C．Nevertheless D．Meanwhile22.A．curious B．controversial C．concerned D．optimistic23.A．In brief B．By contrast C．In addition D．Without doubt 24.A．engagement B．concentration C．puzzle D．definition25.A．memorize B．prioritize C．modernize D．fertilize26.A．representative B．argumentative C．executive D．sensitive27.A．harsh B．adaptable C．crucial D．rare28.A．cheers B．disappoints C．stabilizes D．improves29.A．account for B．count on C．suffer from D．hold up30.A．pouring B．admitting C．integrating D．tempting31.A．pretend B．afford C．offer D．tend32.A．effects B．delights C．intervals D．codes33.A．companions B．scales C．expectations D．findings34.A．necessity B．involvement C．perseverance D．reluctance35.A．equivalent B．exceptional C．mysterious D．distressing Montessori was born in Italy in 1870 with progressive parents, who frequently communicated with the country’s leading thinkers and scholars. This enlightened family environment provided Montessori with many advantages over other young girls of the time.Her mother’s support was vital for some impo rtant decisions, such as her enrolment in a technical school after her elementary education. Her parents’ support also proved to be essential for her decision to study medicine, a field that was dominated by men.Soon after graduating, in 1896, Montessori began work as a voluntary assistant in a clinic at the University of Rome, where she cared for children with learning difficulties. The rooms were bare, with just a few pieces of furniture. One day, she found that the children were enthusiastically playing with breadcrumbs (面包屑) that had dropped on the floor. It then occurred to her that the origin of some intellectual disabilities could be related with poverty. With the right learning materials, these and other young minds could be nurtured, Montessori concluded.The observation would lead Montessori to develop a new method of education that focused on providing optimal stimulation during the sensitive periods of childhood.At its centre was the principle that all the learning materials should be child-sized and designed to appeal to all the senses. In addition, each child should also be allowed to move and act freely, and use their creativity and problem-solving skills. Teachers took the role of guides, supporting the children without press or control.Mont essori opened her first Children’s House in 1907. When the Fascists (法西斯主义者) first came into power in Italy in 1922, they initially embraced her movement. But they soon came to oppose the emphasis on the children’s freedom of expression. Montessori’s value s had always been about human respect, and the rights of children and women, but the Fascists wanted to use her work and her fame.Things reached a breaking point when the Fascist tried to influence the schools’ educational content, and in 1934 Montessori and her son decided to leave Italy. She didn’t return to her homeland until 1947, and she continued to write about and develop her method until her death in 1952, at the age of 81.36. The primary reason for Montessori to develop a new educational method was ______.A．her family’s supportive influence on her educationB．her experience as a voluntary assistant in a clinicC．her observation of children playing with breadcrumbs happilyD．her decision to study medicine, a field dominated by men37. What was a central principle of Montessori’s educational method as described in the passage?A．Providing standardized, one-size-fits-all learning materials.B．Encouraging strict discipline and control over children’s actions.C．Focusing on rote memorization and competition.D．Creating a free and children-centered learning environment.38. Montessori decided to leave Italy in 1934 because .A．she wanted to explore other countries and culturesB．she wanted to avoid the Fascist’s influence on her workC．she was offered a better job in a different countryD．she wanted to retire and enjoy a peaceful life in another country39. Which of the following words can best describe Montessori in this passage?A．Observant and innovative. B．Traditional and emotional.C．Progressive and dependent. D．Open-minded and indifferent. Reducing the workweek to four days could have a climate benefit. In addition to improving the well-being of workers, cutting working hours may reduce carbon emissions. But those benefits would depend on a number of factors, experts emphasize, including how people choose to spend nonworking time.Commuting and travelTransportation is the biggest contributor to greenhouse emissions. A November 2021 survey of2,000 employees and 500 business leaders in the United Kingdom found that if all organizations introduced a four-day week, the reduced trips to work would decrease travel overall by more than 691 million miles a week.But the climate benefits of less commuting could be eliminated, experts said, if people choose to spend their extra time off traveling, particularly if they do so by car or plane.Energy usageShorter working hours could lead to reductions in energy usage, experts said. According to a 2006 paper, if the United States adopted European work standards, the country would consume about 20 percent less energy.Energy could also be conserved if fewer resources are needed to heat and cool large office buildings, reducing demands on electricity. For example, if an entire workplace shuts down on the fifth day, that would help lower consumption — less so if the office stays open to accommodate employees taking different days off.Lifestyle changesIt’s possible that fewer working hours may lead some people to have a larger carbon footprint, bu t experts say research suggests that most people are likely to shift toward more sustainable lifestyles.One theory is that people who work more and have less free time tend to do things in more carbon-intensive ways, such as choosing faster modes of transportation or buying prepared foods. Convenience is often carbon-intensive and people tend to choose convenience when they're time-stressed. Meanwhile, some research suggests that those who work less are more likely to engage in traditionally low-carbon activities, such as spending time with family or sleeping.“When we talk about the four-day workweek and the environment, we focus on the tangible, but actually, in a way, the biggest potential benefit here is in the intangible,” experts said.40. What is identified as the leading cause of greenhouse emissions according to the passage?A．The well-being of employees.B．The conservation of energy.C．Commuting and travel.D．The European work standard.41. What can be inferred from the underlined sentence “the biggest potential benefit here is in the intangible” in the last paragraph?A．People will have big potential in achieving intangible benefits while working.B．People are more likely to engage in carbon-intensive activities due to time constraints.C．People may shift toward more sustainable lifestyles and lower carbon footprints.D．People may travel more frequently by car or plane during their extra time off.42. The passage is mainly written to .A．highlight the importance of shortening working time in the context of well-beingB．provide an overview of transportation emissions worldwideC．analyze the impact of reduced working hours on mode of businessD．illustrate factors affecting the climate benefits of a shorter workweekThe cultivation of plants by ants is more widespread than previously realized, and has evolved on at least 15 separate occasions.There are more than 200 species of ant in the Americas that farm fungi (真菌) for food, but this trait evolved just once sometime between 45 million and 65 million years ago. Biologists regard the cultivation of fungi by ants as true agriculture appearing earlier than human agriculture because it meets four criteria: the ants plant the fungus, care for it, harvest it and depend on it for food.By contrast, while thousands of ant species are known to have a wide variety of interdependent relationships with plants, none were regarded as true agriculture. But in 2016, Guillaume Chomicki and Susanne Renner at the University of Munich, Germany, discovered that an ant in Fungi cultivates several plants in a way that meets the four criteria for true agriculture.The ants collect the seeds of the plants and place them in cracks in the bark of trees. As the plants grow, they form hollow structures called domain that the ants nest in. The ants defecate (排便) at designated absorptive places in these domain, providing nutrients for the plant. In return, as well as shelter, the plant provides food in the form of fruit juice.This discovery prompted Chomicki and others to review the literature on ant-plant relationships to see if there are other examples of plant cultivation that have been overlooked. “They have never really been looked at in the framework of agriculture,” says Chomicki, who is now at the University of Sheffield in the UK. “It’s definitely widespread.”The team identified 37 examples of tree-living ants that cultivate plants that grow on trees, known as epiphytes (附生植物). By looking at the family trees of the ant species, the team was able to determine on how many occasions plant cultivation evolved and roughly when. Fifteen is a conservative estimate, says Campbell. All the systems evolved relatively recently, around 1million to 3 million years ago, she says.Whether the 37 examples of plant cultivation identified by the team count as true agriculture depends on the definitions used. Not all of the species get food from the plants, but they do rely on them for shelter, which is crucial for ants living in trees, says Campbell. So the team thinks the definition of true agriculture should include shelter as well as food.43. According to biologists, why is ant-fungus cultivation considered as a form of true agriculture?A．Because it occurred earlier than human agriculture.B．Because it fulfills the standards typical of agricultural practices.C．Because it redefines the four criteria for true human agriculture.D．Because it is less common than previously thought.44. What motivated Chomicki and others to review the literature on ant-plant relationships?A．They determined on new family trees of the ant species.B．They overlooked some tree-living ants that provided nutrients for the plants.C．They never studied the ant-plant relationships within the context of agriculture.D．They never identified any an t species that engaged in cultivation of fungi.45. Which of the following statements is supported by the team's findings according to the passage?A．Ants’ cultivation of plants is limited to a few specific species.B．The cultivation of fungi by ants is considered the earliest form of agriculture.C．True agriculture in ants involves only food-related interactions with plants.D．Ants have independently cultivated plants on at least 15 distinct occasions.46. What is the passage mainly about?A．The evolution of ants in the plant kingdom.B．The widespread occurrence of ant-plant cultivation.C．The discovery of a new ant species engaging in agriculture.D．The contrast between ant agriculture and human agriculture.What is the likelihood of you having someone who looks just like you? Would it be a good thing? And if you did have one, would you want to meet them?Consider how often your facial features are used to identify you. Your passport, ID card and driving license all feature your face. 47 You may need your face to unlock your smartphone and possibly even need it to exclude you from being present at a crime scene.The word “doppelgänger” refers to a person who looks the same as you, essentially sharing your features; those that you thought were unique to you and your identity. Not identical twins, as a doppelgänger has no relation to you. The idea originated in German folklore. 48So, let's get real. What are the chances of you having one in the first place? There's said to be a one in 135 chance of an exact match for you existing anywhere in the world, so the chances are pretty low, despite folk wisdom promising you otherwise. And the chances of meeting? The mathematical certainty of finding this particular person is supposedly less than one in a trillion.That said, these statistics may be a good thing. Historically, having a double wasn't always a positive. Back in 1999, an innocent American man, indistinguishable from the real criminal, was sent to prison for robbery, where he stayed for 19 years. 49 . In a different case, a woman in New York was accused of trying to poison her doppelgänger with deadly cheesecake so that she could steal her identity!50 The fascination with doppelgängers may be rooted in historical beliefs that facial resemblance meant they were from the same family or had a common ancestor. It leads to the hope that one day you will meet your lookalike, creating the thrill of a potentially strange meeting. However, as these encounters can be both interesting and disturbing, we understand that after such an experience, you might not want to meet your doppelgänger again.passage in no more than 60 words. Use your own words as far as possible.Competitive CheerleadingOver the years, cheerleading has taken two primary forms: game-time cheerleading and competitive cheerleading. Game-time cheerleaders’ main goal is to entertain the crowd and lead them with team cheers, which should not be considered a sport. However, competitive cheerleading is more than a form of entertainment. It is really a competitive sport.Competitive cheerleading includes lots of physical activity. The majority of the teams require a certain level of tumbling (翻腾运动) ability. It’s a very common thing for gymnasts, so it’s easy for them to go into competitive cheerleading. Usually these cheerleaders integrate lots of their gymnastics experience including their jumps, tumbling, and overall energy. They also perform lifts and throws.Competitive cheerleading is also an activity that is governed by rules under which a winner can be declared. It is awarded points for technique, creativity and sharpness. Usually the more difficult the action is, the better the score is. That’s why cheerleaders are trying to experience great difficulty in their performance. Besides, there is also a strict rule of time. The whole performance has to be completed in less than three minutes and fifteen seconds, during which the cheerleaders are required to stay within a certain area. Any performance beyond the limit of time is invalid.Another reason for the fact that competitive cheerleading is one of the hardest sports is that it has more reported injuries. According to some research, competitive cheerleading is the number one cause of serious sports injuries to women. Generally, these injuries affect all areas of the body, including wrists, shoulders, ankles, head, and neck.There can be no doubt that competitive cheerleading is a sport with professional skills. It should be noted that it is a team sport and even the smallest mistake made by one teammate can bring the score of the entire team down. So without working together to achieve the goal, first place is out of reach. ________________________________________________________________________________ ________________________________________________________________________________ ________________________________________________________________________________ ________________________________________________________________________________ ________________________________________________________________________________ ___________________________________________________________________________52. 如果不好好准备，周五的演讲可能会变得一塌糊涂。

专题一阅读理解[全国卷3年考情分析]题型与题量卷别细微环节理解题推理推断题主旨大意题词义揣测题考情分析从统计表可以看出，高考英语阅读理解的题型设置以细微环节理解题和推理推断题为主，兼顾主旨大意题和词义揣测题。

细微环节理解题相对简洁，而其他三种题型相对较难。

在近两年的考查趋向上，细微环节理解题的答案更加隐藏，叙述含蓄，干脆信息题会越来越少，取而代之的将是事实细微环节题加入很多推理、推断、归纳等元素；推理推断题的难度会适当加大。

本专题将对这四种题型进行递进式的指导。

2024 卷Ⅰ7 5 2 1 卷Ⅱ9 3 2 1 卷Ⅲ9 3 2 12024 卷Ⅰ7 6 1 1 卷Ⅱ 5 6 2 2 卷Ⅲ 6 6 2 12024 卷Ⅰ10 3 1 1卷Ⅱ7 5 1 2卷Ⅲ8 4 1 2第一讲细微环节理解题——定位信息巧比对细微环节理解题在英语高考阅读理解中占了较大的比重，而且此类题型相对比较简洁，只须要依据题干中的关键词，回到原文定位信息区间，稍加比对，就能得出正确答案。

因此，对于这类题目要力求读得快、找得准、答得对，力争不丢分，保住基本分才能得高分。

但有些细微环节理解题由于命题人有意设置障碍，把有用信息分散在文章不同位置，有时又有转折、否定等，因此有些题目须要细致地思索、对比、计算、对上下文关键信息把握和分析。

尽管细微环节理解题相对简洁，但不行掉以轻心。

细微环节理解题常见的考查题型有：干脆信息题、间接信息题、概括细微环节理解题和正误推断题。

一、题型特点要知晓(一)细微环节理解题常见设问方式1．特别疑问句形式。

以when, where, what, which, who, how much/many等疑问词引出的问题。

2．推断是非形式。

含有TRUE/FALSE, NOT true或EXCEPT等的推断是非的问题。

此时要留意题干中是否含有否定词，如not, never等。

3．以“According to ...”开头的提问形式。

核农学报37 卷［10］吕茹洁，商庆银，陈乐，曾勇军，胡水秀，杨秀霞. 基于临界氮浓度的水稻氮素营养诊断研究［J］. 植物营养与肥料学报， 2018， 24（5）：1396-1405［11］陆军胜，耿晨鸣，崔晓路，李梦月，胡田田. 基于叶面积指数的夏玉米叶片临界氮浓度稀释曲线研究［J］. 农业机械学报， 2021，52（4）： 319-326［12］苏文楠，解君，韩娟，刘铁宁，韩清芳. 夏玉米不同部位干物质临界氮浓度稀释曲线的构建及对产量的估计［J］. 作物学报，2021， 47（3）： 530-545［13］刘秋霞，任涛，张亚伟，廖世鹏，李小坤，丛日环，鲁剑巍. 华中区域直播冬油菜临界氮浓度稀释曲线的建立与应用［J］. 中国农业科学， 2019， 52（16）： 2835-2844［14］张娟娟，杜盼，郭建彪，曹锐，张捷，马新明. 不同氮效率小麦品种临界氮浓度模型与营养诊断研究［J］. 麦类作物学报， 2017， 37（11）：1480-1488［15］石小虎，蔡焕杰. 基于叶片SPAD估算不同水氮处理下温室番茄氮营养指数［J］. 农业工程学报， 2018， 34（17）： 116-126［16］张加康，李斐，李跃进，杨海波，贾禹泽，刘玉峰，石焱. 基于全株生物量和全株氮浓度的马铃薯氮临界浓度稀释模型的构建及验证［J］. 植物营养与肥料学报， 2020， 26（9）： 1691-1701［17］陈艺文，李用财，余凌羿，王叶琼，倪洪涛. 中国三大主产区甜菜糖业发展分析［J］. 中国糖料， 2017， 39（4）： 74-76， 80［18］王荣华，李守明，艾依肯，丁兆斐，林明. 新疆甜菜产业发展现状与展望［J］. 中国糖料， 2022， 44（1）： 81-86［19］王健，陈花山，王宝，吴子毅，张思聪，赵抒娜. 我国精炼糖厂发展现状、问题及对策［J］. 中国糖料， 2020， 42（3）： 73-80［20］李文生. 塔城盆地地膜甜菜亩产4-5吨、含糖17.0%栽培技术规程［J］. 农村科技， 1995（10）： 12-13［21］张加康，李斐，史树德，杨海波. 内蒙古地区甜菜临界氮浓度稀释模型的构建及应用［J］. 作物学报， 2022， 48（2）： 488-496［22］安志超，黄玉芳，汪洋，赵亚南，岳松华，师海斌，叶优良. 不同氮效率夏玉米临界氮浓度稀释模型与氮营养诊断［J］. 植物营养与肥料学报， 2019， 25（1）： 123-133［23］鲍士旦. 土壤农化分析［M］. 北京：中国农业出版社， 2007［24］Justes E，Mary B，Meynard J M，Machet J M，Thelier-Huche L.Determination of a critical nitrogen dilution curve for winter wheatcrops［J］. 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Five N application rates ［0 （N0），75 （N75）， 150 （N150）， 225 （N225） and 300 kg·hm-2（N300）］ were set. The biomass and N concentration of plant in seedling stage，root tuber formation and differentiation stage，leaf cluster rapid growth stage，root tuber expansion stage， sugar accumulation stage and harvest stage were analyzed. The critical N concentration model based on biomass were developed.The N nutrition index （NNI）was calculated，and its relationship with relative biomass and relative yield were analyzed， respectively， and then the N nutrition status of sugar beet was determined.The results showed the plant biomass increased significantly when N application rate increased from 0 to 225 kg·hm-2， while it did not increase significantly when N application rate increased from 225 to 300 kg·hm-2. From the seedling stage to harvest stage， the plant biomass of N225 treatment increased by 82.55%， and that of N300 treatment increased by 86.44%， respectively， compared with N0 treatment. The variation of plant biomass with different N application rates during the whole growth stage of sugar beet followed the order of N0<N75<N150<N225≈N300. The determination coefficient R2 of critical N concentration model of sugar beet based on biomass was 0.974 and the test parameter n-RMSE was 16.26%， which showed good stability of the model. The recommended N application rate based on NNI was 150~225 kg·hm-2. A significant correlationship was found between NNI and relative biomass， relative yield. In conclusion， the developed critical N concentration model and NNI could be used as a diagnostic tool to detect N nutrition and a theoretical basis to manage nitrogen fertilizer of sugar beet in Ta’e basin， Xinjiang.Keywords：drip-irrigated sugar beet， critical nitrogen dilution curve， nitrogen nutrition index， nitrogen diagnosis核农学报2023，37（5）：1056~1066Journal of Nuclear Agricultural Sciences遮阴时期对不同小麦品种淀粉组成结构及淀粉品质的影响赵佳蓉马宏亮吴东明刘琼樊高琼 *（西南作物基因资源发掘与利用国家重点实验室/四川农业大学农学院/农业农村部西南作物生理生态与耕作重点实验室，四川成都611130）摘要：为研究花前和花后弱光对不同小麦品种淀粉特性的影响，2020—2021年在西昌农科院基地设计二因素裂区试验，主区因素为品种，分别为蜀麦482、川麦39、绵麦51和昌麦34，副区因素为遮阴时期，分别为拔节-开花期（S1）和拔节-成熟期（S2），自然光强为对照（CK）。

胶质瘤是起源于神经上皮的肿瘤，是中枢神经系统最常见的肿瘤［1,2］。

中国国家癌症中心注册中心2015年度报告显示，胶质瘤的发病率和死亡率呈逐年上升的趋势［3,4］。

脑胶质瘤患者的平均生存期小于1年，5年生存率低于5%，目前采用的放疗、化疗、手术切除等多重治疗模式并没有改善胶质瘤患者的预后和生存［5,6］。

因此Lactate-induced up-regulation of PLEKHA4promotes proliferation and apoptosis of human glioma cellsYE Jingjing 1,2,XU Wenqin 1,2,XI Bangsheng 1,3,WANG Nengqian 1,4,CHEN Tianbing 1,21Key Laboratory of Non-coding RNA Transformation Research of Anhui Higher Education Institution,Wannan Medical College,Wuhu 241001,China;2Central Laboratory,3Clinical Laboratory,4Department of Pediatrics,Yijishan Hospital,Wannan Medical College,Wuhu 241001,China摘要：目的探究乳酸诱导的PLEKHA4基因表达对胶质瘤细胞的生物学功能的影响以及潜在分子机制。

方法通过GEO 数据库以及GEPIA2在线网站分析PLEKHA4表达水平与胶质瘤病理分级的关系，通过RNA 小干扰技术设计PLEKHA4siRNA 后分别转染胶质瘤U251和T98G 细胞处理1d ，通过实时细胞分析技术以及Edu 实验检测胶质瘤细胞的增殖能力，平板克隆实验检测细胞的克隆形成能力，流式分析技术检测胶质瘤细胞的周期和凋亡；PCR 检测临床收集的胶质瘤样本和对照以及乳酸和葡萄糖治疗下胶质瘤细胞中PLEKHA4的mRNA 表达量的变化；体内异种移植小鼠实验检测裸鼠成瘤能力；Western blot 检测cyclinD1、CDK2、Bcl2、β-catenin 表达和MAPK 信号通路关键蛋白的磷酸化表达水平。