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股骨头坏死动物模型的分类研究与应用
Chinese Journal of Tissue Engineering Research ｜Vol 25｜No.26｜September 2021｜4211股骨头坏死动物模型的分类研究与应用魏 巍1，楼鹏强1，侯德才2文题释义：动物模型：是指目前实验室中建立的具有人类疾病模拟表现的动物模型，可模拟或者复制人类疾病的病理生理过程。
股骨头坏死动物模型多样化，但并没有统一标准，总结动物模型的优缺点，有利于开展实验研究，并为临床诊疗提供依据。
股骨头坏死：是一种骨质的坏死与修复同时存在的病理演变过程。
人类的股骨头坏死发病率越来越高，要想攻克股骨头坏死这一难题，可使用不同方法在动物实验中寻求突破口，因此构建完美的股骨头坏死模型是问题的关键所在。
摘要背景：目前在基础研究方面，股骨头坏死的模型建立进展较缓慢，并且大多数研究并未阐述各种动物模型的优缺点及优选方法。
目的：探索目前最适合的股骨头坏死动物造模方法及思路，综述各模型的优缺点，并提出目前所存在的问题及解决方法，为以后的疾病研究提供参考及观点依据。
方法：应用计算机检索2009至2020年CNKI 、PubMed 数据库、万方数据知识服务平台，英文检索词 “osteonecrosis of the femoral head ，animal model ，osteoarthritis ，collapse of the femoral head ，trauma ，corticosteroid ，alcohol ，liquid nitrogen ”；中文检索词“股骨头坏死，动物模型，创伤，激素，酒精，液氮”，最终共纳入55篇文献。
结果与结论：①股骨头坏死的常见原因包括酒精、激素以及机械创伤；②其中大鼠无呕吐反射中枢，在酒精型造模中较为常用；③激素引起股骨头坏死的机制尚不明确，主要有脂肪代谢紊乱、血管内凝血和骨质疏松3种假说；动物实验中激素造模的发病机制与人类相似，也可以反映后期股骨头坏死的病理变化；④创伤后股骨头发生软骨下骨折、局部塌陷导致血液循环障碍，引起股骨头缺血坏死；但在动物实验中，创伤型动物的发病机制与人类不相同；⑤但是目前各类动物模型中都存在问题：在激素模型的造模中，激素种类的选取与用量未有统一的标准，模型坏死的部位具有不确定性；犬冷冻模型近几年较受欢迎，但缺乏成熟且精确的操作方法，冷冻范围的不确定对造模后期有一定影响；大部分创伤模型与人类的发病机制不同。
华南农业大学植物生理学2007年考研真题考研试题
页码，1/2考试时间：３小时 满分150分（机 密）华 南 农 业 大 学年攻读硕士学位研究生入学考试试题考试科目：植物生理学用题专业：生态学（考生注意：全部答案必须写在答卷纸上，写在试题上无效。
答案要注明题号，不用抄题。
答卷纸封面需填写自己的考生编号与试题一并交回。
）一、把下列名词先翻译成中文，然后再解释（4×5＝20分）free water Emerson effectphotoperiod grand period of growthplant hormone二、是非判断题并改正（每题2分，共20分，对者打√，错者打╳）1、氮是灰分元素。
( )2、PSⅡ的光反应是短波光反应，其主要特征是ATP的形成。
（）3、叶绿素分子的头部是金属卟啉环，呈极性，因而具有亲水性。
（）4、小麦品种冬性愈强，所需春化温度愈低，春化天数愈短。
( )5、马铃薯块茎，苹果削皮或受伤后出现褐色，就是多酶氧化酶作用的结果。
( )6、ABA具有极性运输的特点。
( )7、生长的最适温度是指生长最快的温度，对健壮生长来说，也是最适宜的。
( )8、抗寒的植物，在低温下合成饱和脂肪酸较多。
( )9、糖酵解是指在线粒体内所发生的，将葡萄糖分解为丙酮酸的过程。
( )10、红光能加速叶片衰老。
( )三、填空题（每空1分，共35分）1、在植物细胞内具有第二信使作用的必需矿质元素是。
2、肉质果实的生长曲线呈型；核果的生长曲线呈型。
3、在光合电子传递中最终电子供体是，最终电子受体是。
2013-07-16。
07-激素(第七章)
（2）性激素
• 性激素属于类固醇类激素，可分为雄性激 素（睾酮）和雌性激素（雌二醇、孕酮） 两类。它们与动物的性别及第二性征的发 育有关。
• 性激素的分泌受垂体的促性腺激素调节。
第20页，共53页。
（三）、脂肪酸衍生物激素
前列腺素
• 前列腺素（简称PG）是一类具有生理活性物质的总称， 现在已发现有几十种，广泛存在于生殖系统和其它组织中， 通过对激素的调节起作用。
• 激素的功能：调节代谢途径；促进细胞和组织的生长；控 制和调节组织器官的生理功能等。
第3页，共53页。
人体内分泌腺
在动物体内，有 些能够分泌激素 的特殊分化细胞 集中在一起构成 内分泌腺；有些 细胞则分散存在； 有些细胞兼具其 他功能。
第4页，共53页。
• 激素被分泌后，可以三种不同的方式作用于靶细胞：
2．信号传递过程： P572 图17－18
H+R
Gs↑
AC↑
cAMP↑
PKA↑
酶蛋白磷酸化
物质代谢改变
糖原分解↑，糖原合成↓ 糖异生↑，糖酵解↓ 脂肪分解↑，脂肪酸合成↓ 胆固醇合成↓ 类固醇激素、儿茶酚胺合成↑
CREB 磷酸化而激活，磷蛋白磷酸酶-1 磷酸化而失活
基因转录表达↑
核蛋白体蛋白质磷酸化
第18页，共53页。
糖皮质激素
• 调节糖代谢：抑制糖的氧化，使血糖升高；促进蛋白质转化 为糖。具有这种功能的包括皮质酮、11-脱氢皮质酮、17-羟 皮质酮（皮质醇、氢化可的松）和17-羟-11-脱氢皮质酮 （可的松）。这类激素还具有良好的抗炎，抗过敏，利尿作 用，是常用的激素药物。
盐皮质激素
• 调节水盐代谢：促使体内保留钠离子及排出过多的钾离子， 调节水盐代谢。这类激素包括醛固酮、17-羟-11-脱氧皮 质酮和11-脱氧皮质酮。
抗缪勒氏管激素的结构、功能、临床应用与检测技术
㊃综述㊃抗缪勒氏管激素的结构㊁功能㊁临床应用与检测技术*林丽淑1,龙韵洪1,徐丽惠1,冯振通1,陈睿1综述,王革非2ә审校(1.深圳市福田区第二人民医院检验科,广东深圳518049;2.汕头大学医学院微生物学与免疫学教研室,广东汕头515041)摘要:抗缪勒氏管激素(AMH)又称缪勒氏管抑制物(M I S),发现于上世纪50年代,是两个相对分子质量为140ˑ103组成的糖蛋白二聚体,属于转化生长因子β超家庭㊂它在胚胎性别分化中起着非常重要作用,参与早期性器官定向分化㊂出生后主要功能为调节男性睾丸间质细胞的功能及参与女性卵泡的规律生成㊂AMH的检测由早期的放射免疫分析技术发展为现在的化学发光技术,在检测灵敏度等方面都有了很大的飞跃㊂其常见临床应用为评估女性的卵巢储备功能㊁诊断早衰㊁多囊卵巢综合征及卵巢颗粒细胞瘤㊁应用于辅助生殖技术等㊂关键词:抗缪勒氏管激素;结构;功能;临床应用;检测技术D O I:10.3969/j.i s s n.1673-4130.2019.24.029中图法分类号:R446.6文章编号:1673-4130(2019)24-3061-07文献标识码:AS t r u c t u r e,f u n c t i o n,c l i n i c a l s i g n i f i c a n c e a n d d e t e c t i o n t e c h n i q u e o f A n t i-Mül l e r i a n h o m o n e*L I N L i s h u1,L O N G Y u n h o n g1,X U L i h u i1,F E N G Z h e n t o n g1,C H E N R u i1,WA N G G e f e i2ә(1.D e p a r t m e n t o f C l i n i c a l L a b o r a t o r y,S e c o n d P e o p l e s H o s p i t a l o f F u t i a nD i s t r i c t,S h e n z h e n,G u a n g d o n g518049,C h i n a;2.D e p a r t m e n t o f M i c r o b i o l o g y a n d I mm u n o l o g y,S h a n t o u U n i v e r s i t y M e d i c a l C o l l e g e,S h a n t o u,G u a n g d o n g515041,C h i n a)A b s t r a c t: A n t i-Mül l e r i a n h o r m o n e(AMH),w h i c h i s a l s o c a l l e d Mül l e r i a n h o r m o n e i n h i b i t i n g s u b s t a n c e (M I S),w a s f o u n d i n t h e1950s.I t i s a g l y c o p r o t e i n w i t h a m o l e c u l a r w e i g h t o f140ˑ103p e p t i d e b e l o n g i n g t o a s u p e r f a m i l y o f g r o w t h f a c t o r s T G F-β.AMH p l a y s a n i m p o r t a n t r o l e i n t h e d i f f e r e n t i a t i o n o f s e x y d e v e l o p m e n t a n d t a k e s p a r t i n t h e s e x u a l o r g a n o r i e n t e d d i f f e r e n t i a t i o n.A f t e r b i r t h,AMH a d j u s t s t h e f u n c t i o n o f L e y d i g c e l l s i n m a l e a n d p a r t i c i p a t e s i n f o l l i c l e d e v e l o p m e n t r e g u l a r l y.AMH w a s d e t e c t e d b y r a d i o i mm u n o a s s a y (R I A)i n t h e p a s t,a n d n o w i s d e t e c t e d b y C h e m i l u m i n e s c e n t i mm u n o a s s a y w i t h h i g h s e n s i t i v i t y.AMH i s u s e d i n e v a l u a t i n g f u n c t i o n o f o v a r y,d i a g n o s i n g p r e m a t u r e o v a r i a n f a i l u r e,p o l y c y s t i c o v a r i a n s y n d r o m e,g r a n u l a r c e l l t u m o r a n d a s s i s t e d r e p r o d u c t i v e t e c h n o l o g y.K e y w o r d s:A n t i-Mül l e r i a n h o m o n e;s t r u c t u r e;f u n c t i o n;c l i n i c a l s i g n i f i c a n c e;d e t e c t i o n t e c h n i q u e抗缪勒氏管激素(AMH)又称缪勒氏管抑制物(M I S)㊂它在胚胎的性别分化中参与早期性器官定向分化[1]㊂在男性睾丸生成过程,AMH于胎儿8周时开始表达,由胎儿睾丸曲精小管的支持细胞生成,诱导男性缪勒氏管退化,而乌尔夫管则发育成男性生殖系统,出生后AMH可调节男性睾丸间质细胞的功能㊂女性AMH于胎儿36周表达,促使缪勒氏管发育成子宫㊁输卵管和阴道的上部分[1]㊂女性AMH的来源主要为卵巢颗粒细胞生成,主要作用机制是通过减少卵泡刺激素(F S H)对窦前和小窦状卵泡的刺激生成作用抑制始基卵泡的募集和窦卵泡的发育,防止卵泡过早耗竭,从而参与卵泡的规律生成[2-3]㊂由此可见,AMH调节生殖管道的分化和发育,对于胚胎早期性别分化至关重要㊂AMH的浓度能准确地反映卵巢的储备功能,比女性年龄㊁抑制素B㊁月经第3天F S H水平㊁雌二醇(E2)㊁B超下窦状卵泡数量(A F C)更灵敏地反映卵巢储备功能,可以诊断卵巢早衰(P O F)㊁应用于辅助生殖技术(A R T)对患者制定个体化促排卵方案㊁诊断多囊卵巢综合征(P O C S)㊁还可作为指导颗粒细胞瘤(G C T)的诊疗手段㊁为其他疾病治疗过程中的女性卵巢储备功能及生育能力评估提供依据[4],以及男性隐睾症与无睾症的区别等[1]㊂可㊃1603㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24*基金项目:国家自然科学基金项目(31170852);广东省自然科学基金项目(2018A030313548)㊂ә通信作者,E-m a i l:g e f e i w a n g@s t u.e d u.c n㊂本文引用格式:林丽淑,龙韵洪,徐丽惠,等.抗缪勒氏管激素的结构㊁功能㊁临床应用与检测技术[J].国际检验医学杂志,2019,40(24): 3061-3066.见,AMH不仅是胚胎性别分化的重要激素,还在女性内分泌激素中扮演着重要角色,还能辅助诊断男性隐睾症与无睾症,是临床某些疾病诊疗重要及可靠的手段㊂1抗缪勒氏管激素的结构AMH是由二硫键连接而成的同源糖蛋白二聚体,相对分子质量140ˑ103,由两个相对分子质量为70ˑ103的单体组成,属于转化生长因子β超家庭(T G F-β),人类AMH基因位于19号染色体短臂(19p l13.3),长2.4~2.8ˑ103,由含5个外显子的275对碱基组成,AMH的基因产物是AMH蛋白前体(p r o-AMH),含560个氨基酸[5]㊂AMH蛋白前体可水解成 可变区 N端(AMH N,相对分子质量115ˑ103)与 成熟区 C端(AMH C,相对分子质量25ˑ103),AMH C端作为活性作用端与受体结合㊂不像其他β转化生长因子,其活化需要AMH N端残基存在[1]㊂AMH颗粒的生物学作用由AMH受体Ⅰ(AMH RⅠ)和AMH受体Ⅱ(AMH RⅡ)的提供,它们存在于间叶细胞㊂AMH R I有激活素受体类激酶A L K2㊁A L K3和A L K63种不同类型㊂AMH与AMH R-I没有直接的亲和力,主要通过AMH RⅡ发挥生物学作用㊂AMH通过与AMHⅡ受体的结合启动与AMH RⅠ受体结合因此形成了二聚体的AMH 复合物,该结合诱导AMH R-I酪氨酸磷酸化导致S m a d蛋白复合体进入细胞核参与基因翻译[1]㊂2抗缪勒氏管激素的生理功能在男性胚胎睾丸生成过程,AMH由胎儿睾丸曲精小管的支持细胞生成,始于胚胎第8周分泌AMH,通过旁分泌作用于缪勒氏管,从而诱导男性缪勒氏管退化,而胚胎另一套生殖管道 乌尔夫管则发育成男性生殖系统(输精管㊁附睾㊁精囊);女性于胚胎第36周卵巢初级卵泡的颗粒细胞开始分泌AMH,缪勒氏管因此得以保留下来并发育成子宫㊁输卵管和阴道的上部分[1]㊂如果男性胚胎的睾丸支持细胞未成分泌AMH或分泌的AMH无生物学活性,或因AMH受体缺陷无法与分泌的AMH结合发挥生物学作用,将会导致如缪勒氏管永存综合征(P M D S)假两性畸形[5]㊂由此可见,AMH参与胚胎早期性器官定向分化,在胚胎的性别早期分化中起着非常重要作用㊂AMH在女性卵巢卵泡发育中起了自分泌与旁分泌的作用,它不表达于始基卵泡,AMH由卵巢初级窦前卵泡和小窦状卵泡的颗粒细胞分泌作用抑制始基卵泡的启动;并且通过抑制细胞色素芳香化酶P450在基因和蛋白质的表达水平[5],减少窦前和小窦卵泡对F S H的依赖性生长,参与生理性卵泡形成过程的始基卵泡募集和优势卵泡募集㊂从而参与优势卵泡的规律生成,是卵泡生长发育的调节因子,防止更多的卵泡消耗,保留卵巢储备[1]㊂免疫组织化学实验结果表明,AMH首先在初级卵泡的颗粒细胞出现,约75%次级卵泡AMH免疫组化呈阳性反应,而最强的免疫组化反应出现于窦前和小窦状卵泡,在人类卵巢,AMH持续表达于卵泡发育直到4~6mm优势卵泡的产生㊂当卵泡发育至大于8mm时,进入F S H 依赖性发育阶段,几乎不分泌AMH,此阶段卵泡发育依赖F S H的分泌[6]㊂可见,AMH表达始于始基卵泡池的募集,不表达于优势卵泡㊂此外,AMH不表达于闭锁卵泡或卵泡膜细胞㊂体外实验证明,无AMH分泌时,初级窦状卵泡细胞到次级窦状卵泡细胞的过渡加强,导致初级卵泡池枯竭㊂小鼠实验还表明,不分泌AMH的鼠窦状卵泡细胞对F S H更敏感,AMH窦状卵泡细胞会降低F S H的阈值,从而增强窦卵泡对F S H的敏感性,增强小窦卵泡的生长发育,在高水平外源性F S H的作用下,卵泡的生长更快,导致更多的卵泡消耗,损耗卵巢储备功能[7]㊂综上所述,AMH主要通过抑制卵泡募集和发育,防止过多的卵泡消耗,使卵泡规律生成,从而发挥保留卵巢储备的作用㊂AMH在人体的生理波动体现在以下方面:在男性,出生前后血清AMH水平出现短暂下降,之后升至高水平,并且持续维持稳定至青春期,随着睾酮分泌的增加,血清AMH迅速下降,并于成年后稳定维持在极低水平[8]㊂男性AMH的生理功能主要表现在调节睾丸间质细胞的功能,睾丸间质细胞受垂体前叶嗜碱性细胞分泌的间质细胞刺激素(黄体生成素)的作用,能合成分泌雄性激素,可促进精子的发生和男性生殖器官发育,以及维持第二性征和性功能㊂女性出生时血清AMH水平远远低于男性出生时的水平,出生后至青春期前AMH血清水平较低并呈复杂的上升趋势,于青春期平均24.5岁到达的AMH峰值,此后随着年龄增长,血清AMH水平逐渐下降,并于更年期急剧下降至极低水平,直到检测不出[6]㊂AMH在不同性别㊁不同年龄段具有不同分泌水平及特点,可以根据监测不同年龄段的AMH水平辅助诊断性早熟及性发育迟缓及㊁诊断卵巢早衰㊁预测更年期等㊂其他影响血清AMH水平的影响还有:种族㊁吸烟㊁维生素D水平与肥胖㊂在美洲,黑人与西班牙女性AMH水平比白种女性低,吸烟女性AMH水平低于不吸烟女性[9],S OW E R S等[10]学者研究表明,吸烟女性比不吸烟女性的AMH水平下降更快㊁闭经年龄更早㊂我国学者研究认为中国女性AMH比白种女性更高,该研究差异是否因研究方法不同原因所导致有待探讨[11]㊂女性血清25-羟维生素D(25-O H-V i t D)的水平与AMH水平呈正相关性,有研究推测维生素D可能作为启动AMH相关基因分泌AMH 的因子㊂在非多囊卵巢综合征女性中,和正常体质量的女性比较,体质量指数(B M I)越高,即越肥胖的女性,其血清AMH及抑制素B(i n h B)水平较B M I低的女性更低,且其A F C具有显著差异,并且肥胖者接受㊃2603㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24I V F的成功率也较正常体质量女性低[12],目前肥胖导致AMH水平下降的机制尚不清楚,在小鼠实验中表明,脂毒性可能为肥胖女性中AMH水平下降的原因[13]㊂AMH的影响因素因不同学者研究人群的抽样方法㊁采用的方法学等不同,研究存在一定差异,对于AMH水平的不同影响因素及不同地区和人种的AMH水平还有待进一步的深入研究㊂3抗缪勒氏管激素的检测技术AMH检测早期方法学为免疫细胞化学检测及放射免疫分析技术(R I A),研究的检测方法主要倾向于AMH在动物组织的定位及功能研究,难以将检测应用及推广至临床诊断㊂AMH水平的检测报道始于上世纪90年代㊂第一代AMH检测为E L I S A检测方法,HU D S O N等[14]及L E E等[15]建立的方法灵敏度分达为0.5n g/m L㊁0.7p m o l/m L㊂商品化试剂由D S L公司和O I T公司研发,他们使用不同的抗体及标准品,检测结果具有较大差异[16]㊂第二代E L I S A AMH检测由贝克曼库尔特公司研发,改良优化了第一代的检测技术,使用了D S L相同的抗体及标准品,其结果大约高于第一代检测方法的22%~40%,但具有较高的相关性[17]㊂接着罗氏也研发了第二代E L I S A AMH检测方法,其检测限(L O Q)达0.5 p m o l/L[18],研究进一步质疑血清标本在储存期间的稳定㊁稀释甚至标本的检测㊂针对有研究者发现的包括一些至2012年已取得体外诊断许可证书的第二代检测试剂盒AMH检测水平低于第一代检测试剂盒[19]㊂2013年7月贝克曼库尔特引进了检测前使用合适缓冲液孵育的方案,这种稀释方案可以避免补体蛋白的结合效应㊂另外在采集2h内冰冻血清可提高检测准确性[20]㊂A n s L a b s公司研发的最新一代检测试剂盒如2012年的超灵敏检测试剂㊁2013年的P i-c o AMH试剂盒,比第二代E L I S A试剂盒具有更高的检测灵敏度和检测限(0.08n g/m L),利于低水平AMH的检出[21]㊂可见,对于第二代检测试剂盒,对于不同厂家,结果的比较和结合临床重要信息显得非常重要[22]㊂2014年罗氏公司率先研发了全自动电化学发光检测AMH,并且指出了不同年龄段女性的参考区间和P C O S的诊断临界值,标志AMH全自动检测的临床诊断应用的开始[23]㊂紧接着贝克曼库尔特也推出了全自动A c c e s s AMH检测系统[24]㊂V A N 等[25]将罗氏公司和贝克曼公司的全自动检测系统与第二代E L I S A检测方法相比,具有较好的相关性, A c c e s s AMH和E l e c s y s AMH检测方法与第二代E L I S A检测方法的偏差分别为9%及12%,前者的检测结果比第二代E L I S A检测方法的结果稍低[26],但有很好的一致性[27],罗氏公司和贝克曼公司全自动检测系统具有检测误差较小及分析灵敏度较高㊁样品稳定性好的优点,为AMH应用于临床检测提供了可靠的实验手段,为大多数用户所选择㊂也是目前国内外较为常用的检测系统㊂4抗缪勒氏管激素的临床应用4.1评估卵巢储备功能女性血清AMH水平可作为卵巢功能独特的内分泌指标,与性类固醇激素㊁促性腺激素和多肽类如抑制素B相比,血清AMH不随月经周期的变化而变化,可以于月经周期的任意时间检测[28],虽然有研究表明女性AMH于黄体早期水平较低,但其在月经周期波动程度比其他性激素相比显得微不足道,并且不受怀孕与服用避孕药物的影响[1]㊂AMH与超声下的窦卵泡计数(A F C)呈正相关性,在作为预测闭经的血清学标志物,AMH的因此可以作为替代超声评估卵巢储备功能更经济㊁更方便的手段㊂可见血清A HM水平是比患者年龄㊁月经第3天的F S H水平㊁雌二醇水平㊁i n h B水平㊁A F C更灵敏㊁更特异的诊断卵巢储备功能的指标,能准确可靠评估卵巢储备功能㊁作为女性生育能力的标志物[28]㊂AMH是目前女性卵巢储备功能较为可靠及稳定的血清学指标,无需指定时间即可检测,联合睾酮及胰岛素的检测对于疾病的早期诊断及治疗有积极的意义,也能诊断胰岛素抵抗性P C O S㊂在肿瘤患者当中,其治疗手段如放疗与化疗与手术均有影响其卵巢储备功能的可能㊂化疗通过对卵泡的不良反应,使化疗后的女性AMH水平显著低于同龄健康女性,与其他药物的化疗相比,用烷化剂治疗后AMH水平显著降低甚至检测不到,化疗后AMH水平的升高取决于治疗方案的选择[29],化疗后卵巢储备功能评估可通过检测血清AMH水平实现[30]㊂有研究表明,儿童肿瘤患者采用放疗与化疗治疗后的幸存者,十年后其血清AMH水平远低于健康同龄人,且患不孕症的比例更高[31]㊂肿瘤患者血清AMH水平在化疗后恢复到化疗前水平的概率与速度与化疗造成的卵巢损害成反比[32]㊂在乳腺癌患者的一项前瞻性研究表明,其术前或放疗㊁化疗前血清AMH水平越低,术后1年内闭经的概率越高,恢复生育能力的概率越低[33]㊂在卵巢囊肿手术中,不同的手术方式对卵巢储备功能的影响不同,在剥除术中选用电凝术止血比选用普通止血材料或缝合方式止血其血清AMH水平下降得更低[34]㊂可见,不同的治疗手段对肿瘤患者血清AMH水平的影响不同,通过AMH预测其卵巢储备功能,有利于帮助不同生育需求的患者选择合适的治疗手段,指导需要保留生育能力的女性选择适当的生育保存程序如冷冻保存胚胎㊁冷冻卵母细胞或卵巢组织以供治疗后移植,为患者后期生育提供了可靠的手段㊂4.2在卵巢早衰的诊断及预测绝经期的应用卵巢早衰(P O F)是常见的女性病症,发病率于1%左右,发病原因主要为原始卵泡储备过少㊁卵泡闭锁或耗竭过快[35]㊂K N A U F F等[36]的研究表明,相比于传统用于诊断卵巢早衰的标志物F S H,有一部分比例患者在发㊃3603㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24生卵巢早衰时,其血清F S H低于诊断切点40I U/ m L,而血清AMH水平对于诊断卵巢早衰的敏感度接近于100%,几乎所有卵巢早衰患者血清AMH水平低于绝经参考区间,说明对于卵巢早衰的诊断大大优于F S H㊂血清AMH水平下降早于血清F S H水平的升高和B超下A F C的下降,成年女性血清AMH水平随年龄增长而逐渐下降,到了更年期,血清AMH水平降到检测不到的极低水平㊂多位学者的多因素研究显示,只有AMH水平可以很好地预测绝经年龄,且比年龄更能准确地预测女性绝经年龄[37],因此血清AMH水平可以用于预测卵巢病理上的衰老和预测绝经年龄[38]㊂还有研究表明如果把年龄㊁月经周期㊁B M I㊁吸烟与否及吸烟量加入AMH进行多因素分析,可以显著提高预测10年内绝经的能力[39]㊂利用AMH诊断卵巢早衰具有敏感度高㊁检测方便㊁诊断效能高等特点,运用多因素分析可提高预测绝经年龄的能力㊂4.3在辅助生殖技术的应用通过AMH在辅助生殖技术(A R T)作用可对患者制定个体化治疗方案,准确合理评估卵巢的储备功能及卵巢对控制性超排卵(C O H)的反应性,提高妊娠率,降低并发症危险因素[40]㊂B R O E R等[41]的M e t a分析表明,血清AMH 水平应用于预测卵巢低反应(P O R)的R O C曲线下面积(0.81)大于年龄㊁月经第3天F S H水平和B超下A F C,临床应用价值更高,N E L S O N等[42]多中心研究表明用AMH预测促排卵人群排卵数比B超下A F C 更有效㊁更显著㊂通过血清AMH水平预测治疗者对激素作用下卵巢刺激(C O S)反应,防止卵巢过度刺激综合征(O H S S),并选择合适治疗方案[43]㊂O H S S是A R T促排卵过程的主要并发症之一,高AMH表示卵巢有过度刺激风险,宜制定小剂量F S H与促性腺激素释放激素(G n R h)拮抗剂治疗方案,低AMH表示卵巢低反应,应制定高剂量F S H治与G n R h拮抗剂疗方案[44-45]㊂有研究显示AMH与I V F出生率相关,I V F出生率与基础AMH水平呈正比㊂一项针对中国人的研究为促排卵人群建立了血清AMH水平用于预测卵巢刺激的切点,AMH1.1n g/m L为发生卵巢低反应的切点,2.6n g/m L为卵巢过度刺激的切点[46]㊂AMH为辅助生殖提供了确切的切点,更精准地应用于促排卵方案的应用,极大提高了患者使用促排卵方案的安全性,减少卵巢过度刺激,为患者促排卵的有效性及身心健康提供了可能依据及保证㊂4.4用于诊断多囊卵巢综合征多囊卵巢综合征(P C O S)是女性表现为以少排卵或无排卵㊁高雄激素或胰岛素抵抗㊁多囊卵巢为特征的内分泌紊乱的症候群,是不孕症的主要病因之一㊂在组织学检查中,P-C O S患者卵巢窦前卵泡数比正常女性高5倍,多囊卵巢的颗粒细胞分泌的AMH比健康排卵女性高75倍[47],有研究表明,P C O S的女性的卵泡液AMH和血清AMH水平比规律排卵的女性分别高5倍与2~ 3倍,血清AMH用于诊断P C O S的特异性和灵敏度分别为92%与67%,并且随着AMH浓度的增高,诊断P C O S的概率越高[48],T A L等[49]的研究表明当AMH>10n g/m L时,P C O S的确诊率可高达97%~ 100%㊂在没有准确的超声数据情况下,AMH水平可以作为替代A F C作为诊断P C O S的标准[50],并有研究表明将AMH水平2.8~8.4n g/m L作为不同人群的P C O S诊断切点值[51]㊂P C O S患者AMH水平的升高可能是由于卵泡发育的中断导致窦前和小窦卵泡累积而分泌过多AMH[50]㊂研究表明,在P C O S患者中,发生胰岛素抵抗的患者血清AMH水平比胰岛素敏感性正常的更高,发生闭经的P C O S患者血清AMH水平比未闭经的更高,提示高水平AMH可以反映闭经卵巢中卵泡发育中断和颗粒细胞功能的受损,可能是P C O S患者提前闭经的原因[51]㊂与B超相比,高水平AMH对于早期P C O S诊断具有方便快捷㊁确认率高的特点,AMH的高确诊率有利于P C O S 的早期诊断及早期治疗㊂4.5 AMH在诊断卵巢肿瘤的应用卵巢颗粒细胞瘤(G C T)发生率占所有卵巢肿瘤的3%~5%,在G C T患者中,76%~93%患者表现出AMH水平的升高[52],在肿瘤复发前16个月即可检测高浓度的AMH水平[1]㊂由此可见,AMH可作为G C T的血清学标志物,并且血清AMH水平与卵巢颗粒细胞肿瘤的大小呈正比关系,与影像学结果具有相关性,对G C T的治疗及复发监测有重要的临床意义[53]㊂基于AMH在肿瘤复发前16个月即可检测到高浓度的事实,提示卵巢颗粒细胞瘤患者定期复查血清AMH水平能更早地发现肿瘤是否复发,有助于患者及时治疗㊂4.6其他应用研究表明,AMH作为类固醇生长的辅助调节因子,存在于颗粒细胞中,在卵泡液,AMH 与E2水平呈对应关系,在人类卵巢AMH在F S H依赖性的类固醇生长起了重要作用[1]㊂对于因促性腺激素分泌过多引起的性腺机能减退,AMH可作为子宫内化学治疗㊁卵巢手术㊁子宫内膜异位症手术等卵巢微小病变的标志物㊂研究实验表明,大量卵巢上皮细胞癌起源于输卵巢管与次级缪勒氏系统,基于AMH可以导致缪勒氏管退化的事实,已有尝试使用AMH来治疗卵巢上皮细胞癌㊂体外研究实验表明AMH可以抑制卵巢癌上皮细胞系的增长[54]㊂国外也有AMH对于卵巢癌其他细胞系的作用研究,这些研究为卵巢癌提供了新思路,AMH作为人体内激素若能应用于卵巢癌治疗,相比较于其他药物治疗或放化疗,具有不良反应相对小的优点㊂在无法触及睾丸的男性,美国泌尿外科学会㊃4603㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24(A U A)指南2014年指出AMH可用于判断区别隐睾症和无睾症,前者血清中检测到的AMH可证明睾丸组织的存在,对于区别无睾症简便快速,可减少隐睾症或无睾丸症的误诊㊂5小结目前国内对于AMH的临床运用主要体现于作为临床疾病的诊疗手段及辅助生殖技术促排卵方案选用的参考依据,但对于AMH蛋白的基础研究基本空白,随着生物技术工程研究的进展,单克隆抗原或抗体治疗技术对肿瘤细胞或相关结构选择选择性识别与破坏能力的提高,近年来许多学者致力于对新型抗卵巢癌药物的研究,以期发现更有效的治疗方案[55]㊂在体内外实验中,高纯度重组人AMH可在体内外抑制乳腺癌㊁前列腺癌㊁宫颈癌㊁子宫内膜癌及卵巢细胞增殖[56-57],笔者近期也做了AMH重组蛋白对卵巢癌S K O V3细胞系增殖凋亡影响的相关研究,初步证明AMH对该细胞系有促凋亡作用,值得进一步研究AMH对卵巢癌的作用,为卵巢癌治疗提供新思路㊂参考文献[1]MA R Z E N A R Z E S Z W S K A,A G N I E S Z K A L E S Z C Z,e ta l.A n t i-Mül l e r i a n h o r m o n e:s t r u c t u r e,p r o p e r t i e s a n d a p-p l i a n c e[J].V i a M e d i c a,2016,87(9):669-674. [2]邱玲,李丹丹,程歆琦.抗缪勒管激素的临床应用[J].协和医学杂志,2017,8(4):229-234.[3]V A N H E L D E N J,W E I S K I R C H E N R.P e r f o r m a n c e o ft h e t w o n e w f u l l y a u t o m a t e d a n t i-Mül l e r i a n h o r m o n e i m-m u n o a s s a y s c o m p a r e d w i t h t h e c l i n i c a l s t a n d a r d a s s a y[J].H u m R e p r o d,2015,30(8):1918-1926. [4]J AM I L Z,F A T I MA S S,A HM E D K,e t a l.A n t i-Mül l e r i a n h o r m o n e:a b o v e a n d b e y o n d c o n v e n t i o n a l o v a r-i a n r e s e r v e m a r k e r s[J].D i s M a r k e r s,2016,20(16):5246217.[5]G R O S S MA N M P,N A K A J I MA S T,F A L L A T M E,e ta l.Mül l e r i a n i n h ib i t i n g s u b s t a nc e i n h i b i t s c y t o c h r o m eP450a r o m a t a z e a c t i v i t y i n h u m a n g r a n u l o s a l u t e i n c e l lc u l t u r e[J].F e r t i l S t e r i l,2008,89(5):1364-1370.[6]B R O E R S L,B R O E KMA N S F J M,L A V E N J S E,e t a l.A n t i-M u l l e r i a n h o r m o n e:o v a r i a n r e s e r v e t e s t i n g a n d i t sp o t e n t i a l c l i n i c a l i m p l i c a t i o n s[J].H u m R e p r o d U p d a, 2014,20(5):688-701.[7]D E WA I L L Y D,A N D E R S E N C Y,B A L E N A,e t a l.T h ep h y s i o l o g y a n d c l i n i c a l u t i l i t y o f a n t i-Mül l e r i a n h o r m o n ei n w o m e n[J].H u m R e p r o d U p d a t e,2014,20(3):370-385.[8]L E E M M,D O N A HO E P K.Mül l e r i a n i n h i b i t i n g s u b-s t a n c e:a g o n a d a l h o r m o n e w i t h m u l t i p l e f u n c t i o n s[J].E n d o c r R e v,1993,14(2):152-164.[9]B L E I L M E,G R E G O R I C H S E,A D L E R N E,e t a l.R a c e/e t h n i c d i s p a r i t i e s i n r e p r o d u c t i v e a g e:a n e x a m i n a-t i o n o f o v a r i a n r e s e r v e e s t i m a t e s a c r o s s f o u r r a c e/e t h n i cg r o u p s o f h e a l t h y,r e g u l a r l y c y c l i n g w o m e n[J].F e r t i lS t e r i l,2014,101(1):199-207.[10]S OW E R S M R,M C C O N N E L L D,Y O S E F M,e t.a l.R e-l a t i n g s m o k i n g,o b e s i t y,i n s u l i n r e s i s t a n c e a n d o v a r i a n b i-o m a r k e r c h a n g e s t o t h e f i n a l m e n s t r u a l p e r i o d(F M P) [J].A n n N Y A c a d S c i,2010,1204(1):95-103. [11]D U X,D I N G T,Z HA N G H,e t a l.A g e-s p e c i f i c n o r m a lr e f e r e n c e r a n g e f o r s e r u m a n t i-mül l e r i a n h o r m o n e i nh e a l t h y c h i n e s e h a n w o m e n:a n a t i o n w i d e p o p u l a t i o n-b a s e d s t u d y[J].R e p r o d Sc i,2016,23(8):1019-1027.[12]F R E E MA N E,G R A C I A C,S AMM E L M,e t a l.A s s o c i a-t i o n o f a n t i-Mül l e r i a n h o r m o n e l e v e l s w i t h o b e s i t y i n l a t e r e p r o d u c t i v e a g e w o m a n[J].F e r t i l S t e r i l,2007,87(1): 101-106.[13]WU L L Y,D U N N I N G K R,Y A N G X,e t a l.H i g h t-f a t d i-e t c a u s e s l i p o t o x i c i t y r e s p o n s e s i n c u m u l u s o o c y t e c o m-p l e x e s a n d d e c r e a s e d f e r t i l i z a t i o n r a t e s[J].E n d o c r i n o l, 2010,151(11):5438-5445.[14]HU D S O N P L,D O U G A S I,D O N A HO E P K,e t a l.A ni mm u n o a s s a y t o d e t e c t h u m a n m u l l e r i a n i n h i b i t i n g s u b-s t a n c e i n m a l e s a n d f e m a l e s d u r i n g n o r m a l d e v e l o p m e n t [J].J C l i n E n d o c r i n o l M e t a b,1990,70(1):16-22. [15]L E E M M,D O N A HO E P K,HA S E G AWA L,e t a l.M u l-l e r i a n i n h i b i t i n g s u b s t a n c e i n h u m a n s:n o r m a l l e v e l s f r o mi n f a n c y t o a d u l t h o o d[J].J C l i n E n d o c r i n o l M e t a b,1996,81(2):571-576.[16]B R O E R S L,B R O E KMA N S F J,L A V E N J S,e t a l.A n t i-Mül l e r i a n h o r m o n e:o v a r i a n r e s e r v e t e s t i n g a n d i t s p o t e n-t i a l c l i n i c a l a p p l i c a t i o n s[J].H u m R e p r o d U p d a t e,2014, 20(1):688-701.[17]WA L L A C E A M,F A Y E S A,F L E M I N G R,e t a l.Am u l t i c e n t r e e v a l u a t i o n o f t h e n e w B e c k m a n C o u l t e r a n t i-M u l l e r i a n h o r m o n e i mm u n o a s s a y(AMH G e nⅡ)[J].A n n C l i nB i o c h e m,2011,48(14):370-373.[18]H Y L D G A A R D J,B O R P,I N G E R S L E V H J,e t a l.C o m-p a r i s o n o f t w o d i f f e r e n t m e t h o d s f o r m e a s u r i n g a n t i-m u l-l e r i a n h o r m o n e i n a c l i n i c a l s e r i e s[J].R e p r o d B i o l E n d o-c r i n o l,2015,13(1):1-4.[19]L U K A S Z U K K,L U DW I K OW S K A B,L I S S J,e t a l.D e-c r e a s i n g q u a l i t y o f t h e n e w g e n e r a t i o n s o f a n t i-Mül l e r i a nh o r m o n e a s s a y s[J].B i o m e d R e s I n t,2014,20(14):165352.[20]V I S S E R J A,L A V E N J S,M C L U S K E Y A,e t a l.D e v e l-o p m e n t o f a w e l l c h a r a c t e r i z e d u l t r a.s e n s i t i v e h u m a n a n-t i-m u l l e r i a n h o r m o n e c h e m i l u m i n e s c e n c e a s s a y:e v a l u a-t i o n o f p o t e n t i a l c l i n i c a l a p p l i c a t i o n s[C].E n d o c r i n e S o c i e-t y A n n u a l M e e t i n g,2013.[21]W E L S H P,S M I T H K,N E L S O N S M.A s i n g l e-c e n t r e e-v a l u a t i o n t o t w o n e w a n t i-Mül l e r i a n h o r m o n e a s s a y s a n d c o m p a r i s o n w i t h t h e c u r r e n t c l i n i c a l s t a n d a r d a s s a y[J].H u m R e p r o d,2014,29(1):1035-1041.[22]A T T A E L MA N N A N M,P A N D I A N R,S A D R I L M,e ta l.D e v e l o p m e n t o f ab r o a d-r a n g e,s e n s i t i v e,a n d s p ec i f i ci mm u n o a s s a y f o r a n t i-m u l l e r i a n h o r m o n e(AMH)w i t h n o㊃5603㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24d i l u t i o n i s s ue s[C].A m e r i c a n A s s o c i a t i o nf o r C l i n i c a lC h e m i s t r y A n n u a1M e e t i n g,2013.[23]G A S S N E R D,J U N G R.F i r s t f u l l y a u t o m a t e d i mm u n o a s-s a y f o r a n t i-Mül l e r i a n h o r m o n e[J].C l i n C h e m L a b M e d, 2014,52(8):1143-1152.[24]D E M I R D J I A N G,B O R D S,L E J E U N E C,e t a l.P e r f o r m-a n c e c h a r a c t e r i s t i c s o f t h e A c c e s s AMH a s s a y f o r t h eq u a n t i t a t i v e d e t e r m i n a t i o n o f a n t i-M i i l l e r i a n h o r m o n e (AMH)l e v e l s o n t h e A c c e s s'f a m i l y o f a u t o m a t e d i mm u-n o a s s a y s y s t e m s[J].C l i n B i o c h e m,2016,49(16/17): 1267-1273.[25]V A N H J,W E I S K I R C H E N R.P e r f o r m a n c e o f t h e t w on e w f u l l y a u t o m a t e d a n t i-M u l l e r i a n h o r m o n e i mm u n o a s-s a y s c o m p a r e d w i t h t h e c l i n i c a l s t a n d a r d a s s a y[J].H u mR e p r o d,2015,30(8):1918-1926.[26]N E L S O N S M,P A S T U S Z E K E,K L O S S G,e t a l.T w on e w a u t o m a t e d,c o m p a r e d w i t h t w o e n z y m e-l i n k e d i mm u-n o s o r b e n t,a n t i mül l e r i a n h o r m o n e a s s a y s[J].F e r t i l S t e r-i l,2015,104(1):1016-1021.[27]P E A R S O N K,L O N G M,P R A S A D J,e t a l.A s s e s s m e n t o f t h e a c c e s s AMH a s s a y a s a n a u t o m a t e d,h i g h-p e r f o r m-a n c e r e p l a c e m e n t f o r t h e AMH G e n e r a t i o nⅡm a n u a lE L I S A[J].R e p r o d B i o l E n d o c r i n o l,2016,14(1):8.[28]N E L S O N S M,K L E I N B M,A R C E J C.C o m p a r i s o n o fa n t i mül l e r i a n h o r m o n e l e v e l s a n d a n t r a l f o l l i c l e c o u n t a sp r e d i c t o r o f o v a r i a n r e s p o n s e t o c o n t r o l l e d o v a r i a n s t i m u-l a t i o n i n g o o d-p r o g n o s i s p a t i e n t s a t i n d i v i d u a l f e r t i l i t y c l i n i c s i n t w o m u l t i c e n t e r t r i a l s[J].F e r t i l i t y a n d S t e r i l i t y,2015,103(4):923-931.[29]P E I G NÉM,D E C A N T E R C.S e r u m AMH l e v e l a s am a r k e r o f a c u t e a n d l o n g-t e r m e f f e c t s o f c h e m o t h e r a p y o n t h e o v a r i a n f o l l i c u l a r c o n t e n t:a s y s t e m a t i c r e v i e w[J].R e p r o d B i o l E n d o c r i n o l,2014,12(1):26. [30]H E N R Y N L,X I A R,S C HO T T A F,e t a l.P r e d i c t i o n o fp o s t c h e m o t h e r a p y o v a r i a n f u n c t i o n u s i n g m a r k e r s o f o-v a r i a n r e s e r v e[J].O n c o l o g i s t,2014,19(1):68-74. [31]M E R T E N S A C,Y O N G J,D I E T Z A C,e t a l.C o n d i t i o n a l s u r v i v a l i n p e d i a t r i c m a l i g n a n c i e s:a n a l y s i s o f d a t a f r o m t h e c h i l d h o o d c a n c e r s u r v i v o r s t u d y a n d t h e s u r v e i l l a n c e, e p i d e m i o l o g y,a n d e n d r e s u l t s p r o g r a m[J].C a n c e r,2015, 121(1):1108-1117.[32]I WA S E A,N A K AMU R A T,N A K A HA R A T,e t a l.A n-t i-M u l l e r i a n h o r m o n e a n d a s s e s s m e n t o f o v a r i a n r e s e r v e a f t e r o v a r i a n t o x i c t r e a t m e n t:a s y s t e m a t i c n a r r a t i v e r e-v i e w[J].R e p r o d S c i,2015,22(1):519-526. [33]B A R N A B E I A,S T R I G A R I L,MA R C H E T T I P,e t a l.P r e d i c t i n g o v a r i a n a c t i v i t y i n w o m e n a f f e c t e d b y e a r l yb r e a s tc a n c e r:A M e t a a n a l y s i s-b a s ed n o m o g r a m[J].O n-c o l o g i s t,2015,20(1):1111-1118.[34]I WA S E A,N A K AMU R A T,N A K A HA R A T,e t a l.A s-s e s s m e n t o f o v a r i a n r e s e r v e u s i n g a n t i-Mül l e r i a n h o r m o n e l e v e l s i n b e n i g n g y n e c o l o g i c c o n d i t i o n s a n d s u r g i c a l i n t e r-v e n t i o n s:a s y s t e m a t i c n a r r a t i v e r e v i e w[J].R e p r o d B i o lE n d o c r i n o,2014,12(1):125.[35]B R O E R S L,E I J K E MA N S M J,S C H EF F E RG J,e t a l.A n t i-mül l e r i a n h o r m o n e p r e d i c t s m e n o p a u s e:a l o n g-t e r m f o l l o w-u p s t u d y i n n o r m o o v u l a t o r y w o m e n[J].J C l i n E n-d o c r i n o l Me t a b,2011,96(8):2532-2539.[36]K N A U F F E A,E I J K E MA N S M J,L AM B A L K C B,e ta l.A n t i-Mül l e r i a n h o r m o n e,i n h ib i n B,a n d a n t r a l f o l l ic l ec o u n t i n y o u n g w o m e n w i t h o v a r i a n f a i l u r e[J].J C l i n E n-d o c r i n o l Me t a b,2009,94(1):786-792.[37]D O L L E MA N M,D E P MA N N M,E I J K E MA N S M J,e ta l.A n t i-Mül l e r i a n h o r m o n e i s a m o r e a c c u r a t e p r e d i c t o ro f i n d i v i d u a l t i m e t o m e n o p a u s e t h a n m o t h e r's a g e a t m e n o p a u s e[J].H u m R e p r o d,2014,29(1):584-591.[38]D O L L E MA N M,D E P MA N N M,E I J K E MA N S M J,e ta l.A n t i-Mül l e r i a n h o r m o n e i s a m o r e a c c u r a t e p r e d i c t o ro f i n d i v i d u a l t i m e t o m e n o p a u s e t h a n m o t h e r's a g e a tm e n o p a u s e[J].H u m R e p r o d,2014,29(3):584-591.[39]D O L L E MA N M,V E R S C HU R E N W M,E I J K E MA N SM J,e t a l.A d d e d v a l u e o f a n t i-Mül l e r i a n h o r m o n e i n p r e-d i c t i o n o f me n o p a u s e:e s u l t sf r o m a l a rg e p r o s p e c t i v e c o-h o r t s t u d y[J].H u m R e p r o d,2015,30(8):1974-1981.[40]蒋其,许耀辉,李娜,等.AMH水平预测P C O S患者行辅助生殖技术超促排卵结果的价值[J].临床输血与检验, 2019,20(5):531-534.[41]B R O E R S L,V A N D I S S E L D O R P J,B R O E Z E K A,e t a l.A d d e d v a l u e o f o v a r i a n r e s e r v e t e s t i n g o n p a t i e n t c h a r a c-t e r i s t i c s i n t h e p r e d i c t i o n o f o v a r i a n r e s p o n s e a n d o n g o i n g p r e g n a n c y:a n i n d i v i d u a l p a t i e n t d a t a a p p r o a c h[J].H u m R e p r o d U p d a t e,2013,19(1):26-36.[42]N E L S O N S M,K L E I N B M,A R C E J C.C o m p a r i s o n o fa n t i Mül l e r i a n h o r m o n e l e v e l s a n d a n t r a l f o l l i c l e c o u n t a sp r e d i c t o r o f o v a r i a n r e s p o n s e t o c o n t r o l l e d o v a r i a n s t i m u-l a t i o n i n g o o d p r o g n o s i s p a t i e n t s a t i n d i v i d u a l f e r t i l i t y c l i n-i c s i n t w o m u l t i c e n t e r t r i a l s[J].F e r t i l S t e r i l,2015,103(4):923-930.[43]L A MA R C A A,S U N K A R A S K.I n d i v i d u a l i z a t i o n o fc o n t r o l l ed o v a r i a n s t i m u l a t i o n i n I V F u s i n g o v a r i a n r e-s e r v e m a r k e r s:f r o m t h e o r y t o p r a c t i c e[J].H u m R e p r o dU p d a t e,2014,20(1):124-140.[44]R E I C HMA N D E,G O L D S C H L A G D,R O S E NWA K S Z.V a l u e o f a n t i mül l e r i a n h o r m o n e a s a p r o g n o s t i c i n d i c a t o r o f i n v i t r o f e r t i l i z a t i o n o u t c o m e[J].F e r t i l S t e r i l,2014,101(4):1012.[45]L E F E B V R E J,A N T A K I R,K A D O C H I J,e t a l.450I Uv e r s u s600I U g o n a d o t r o p i n f o r c o n t r o l l e d o v a r i a n s t i m u-l a t i o n i n p o o r r e s p o n d e r s:a r a n d o m i z e d c o n t r o l l e d t r i a l [J].F e r t i l i t y a n d S t e r i l i t y,2015,104(6):1419-1425.[46]L I R,G O N G F,Z HU Y,e t a l.A n t i-Mül l e r i a n h o r m o n ef o r p r e d i c t i o n o f o v a r i a n r e s p o n s e i n C h i n e s e i n f e r t i l ew o m e n u n d e r g o i n g I V F/I C S I c y c l e s:a p r o s p e c t i v e,m u l t i-c e n t r e,o b s e r v a t i o n a l s t ud y[J].Re p r o d B i o M e d O n l i n e,2016,33(4):506-512.[47]D E WA I L L Y D.D i a g n o s t i c c r i t e r i a f o r P C O S:I s t h e r e an e e d f o r a r e t h i n k[J].B e s t P r a c t R e s C l i n O b s t e t G y n a e-c o l,2016,37(1):5-11.(下转第3072页)㊃6603㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24者[10]㊂该患者因高龄及经济因素,家属放弃化疗及靶向治疗,采取姑息支持治疗,目前已存活7年㊂3小结综上所述,L P L是一种少见的惰性成熟B细胞淋巴瘤,随着分子遗传学的进一步研究及新药的研制, L P L的诊疗方法将更加完善㊂研究者首次报道1例I g A和I g M双克隆型L P L,由于该病极其罕见,今后的临床实践中需要进一步积累资料,提高对该病的认识水平㊂参考文献[1]中国抗癌协会血液肿瘤专业委员会,中华医学会血液学分会白血病淋巴瘤学组,中国抗淋巴瘤联盟.淋巴浆细胞淋巴瘤/华氏巨球蛋白血症诊断与治疗中国专家共识(2016年版)[J].中华血液学杂志,2016,37(9):729-734.[2]Y I S H,C U I R,L I Z J,e t a l.D i s t i n c t c h a r a c t e r i s t i c s a n dn e w p r o g n o s t i c s c o r i n g s y s t e m f o r C h i n e s e p a t i e n t s w i t hW a l d e n s t röm m a c r o g l o b u l i n e m i a[J].C h i n M e d J,2014, 127(12):2327-2331.[3]邹德慧,易树华,刘慧敏,等.非I g M型淋巴浆细胞淋巴瘤临床及生物学特征研究[J].中华血液学杂志,2015,36(6):493-496.[4]C A O X,M E D E I R O S L J,X I A Y,e t a l.C l i n i c o p a t h o l o g i cf e a t u r e s a n d o u t c o m e s o f l y m p h o p l a s m a c y t i c l y m p h o m ap a t i e n t s w i t h m o n o c l o n a l I g G o r I g A p a r a p r o t e i n e x p r e s-s i o n[J].L e u k L y m p h o m a,2016,57(5):1104-1113. [5]王继军,景红梅,申红卫,等.16例淋巴浆细胞淋巴瘤/华氏巨球蛋白血症临床特点[J].中国实验血液学杂志, 2010,18(6):1494-1498.[6]G E R T Z M A.W a l d e n s t röm m a c r o g l o b u l i n e m i a:2017u p-d a te o n d i a g n o s i s,r i s k s t r a t if i c a t i o n,a n d m a n ag e m e n t[J].A m J H e m a t o l,2017,92(2):209-217.[7]K I N G R L,G O N S A L V E S W I,A N S E L L S M,e t a l.L y m p h o p l a s m a c y t i c l y m p h o m a w i t h a n o n-I g M p a r a p r o-t e i n s h o w s c l i n i c a l a n d p a t h o l o g i c h e t e r o g e n e i t y a n d M a yH a r b o r MY D88L265P[J].A m J C l i n P a t h o l,2016,145(6):843-851.[8]C HA K R A B O R T Y R,K A P O O R P,A N S E L L S M,e t a l.E m e r g i n g t h e r a p e u t i c o p t i o n s f o r W a l d e n s t röm m a c r o-g l o b u l i n e m i a/l y m p h o p l a s m a c y t i c l y m p h o m a[J].E x p e r tR e v A n t i c a n c e r T h e r,2015,15(10):1143-56.[9]G E R T Z M A.W a l d e n s t röm m a c r o g l o b u l i n e m i a t r e a t m e na l g o r i t h m2018[J].B l o o d C a n c e r J,2018,8(4):40.[10]L E B L O N D V,K A S T R I T I S E,A D V A N I R,e t a l.T r e a t-m e n t r e c o mm e n d a t i o n s f r o m t h e E i g h t h I n t e r n a t i o n a l W o r k s h o p o n W a l d e n s t röm's M a c r o g l o b u l i n e m i a[J].B l o o d,2016,128(10):1321-1328.(收稿日期:2019-05-14修回日期:2019-09-15)(上接第3066页)[48]D E WA I L L Y D.D i a g n o s t i c c r i t e r i a f o r P C O S:I s t h e r e an e e d f o r a r e t h i n k[J].B e s t P r a c t R e s C l i n O b s t e t G y n a e-c o l,2016,37(1):5-11.[49]T A L R,S E I F E R D B,K HA N I MO V M,e t a l.C h a r a c t e r-i z a t i o n o f w o m e n w i t h e l e v a t e d a n t i mül l e r i a n h o r m o n e l e v e l s(AMH):c o r r e l a t i o n o f AMH w i t h p o l y c y s t i c o v a r-i a n s y n d r o m e p h e n o t y p e s a n d a s s i s t e d r e p r o d u c t i v e t e c h-n o l o g y o u t c o m e s[J].A m J O b s t e t G y n e c o l,2014,211(1):59.[50]P I G N Y P,J O N A R D S,R O B E R T Y,e t a l.S e r u m A n t i-Mül l e r i a n h o r m o n e a s a s u r r o g a t e f o r a n t r a l f o l l i c l e c o u n t f o r d e f i n i t i o n o f t h e p o l y c y s t i c o v a r y s y n d r o m e[J].A m JO b s t e t G y n e c o l,2006,91(3):941-945.[51]I L I O D R O M I T I S,K E L S E Y T W,A N D E R S O N R A,e t a l.C a n a n t i-Mül l e r i a n h o r m o n e p r e d i c t t h e d i a g n o s i s o f p o l y c y s-t i c o v a r y s y n d r o m e:A s y s t e m a t i c r e v i e w a n d m e t a-a n a l y s i s o fe x t r a c t e d d a t a[J].J C l i n E n d o c r i n o l M e t a b,2013,98(8):3332-3340.[52]R E Y R A,S A B O U R I N J,V E N A R A M,e t a l.A n t i-Mül l e r i a n h o r m o n e i s s p e c i f i c m a r k e r o f s e r t o l i-a n d g r a n-u l o s e-c e l l o r i g i n i n g o n a d a l t u m o r s[J].H u m P a t h o l,2000,31(10):1202-1203.[53]F A R K K I L A A,K O S K E L A S,B R Y K S,e t a l.T h e c l i n i c a lu t i l i t y o f s e r u m a n t i-Mül l e r i a n h o r m o n e i n t h e f o l l o w-u p o f o v a r i a n a d u l t-t y p e g r a n u l o s a c e l l t u m o r s-a c o m p a r a t i v e s t u d y w i t h i n h i b i n B[J].I n t J C a n c e r,2015,137(7):1661-1671.[54]S T E P H E N A,P E A R S O L L L,C H R I S T I A N B,e t a l.H i g h l y p u r i f i e d Mül l e r i a n i n h i b i t i n g s u b s t a n c e i n h i b i t sh u m a n o v a r i a n c a n c e r i n v i v o[J].C l i n C a n c e r R e s,2002,8(8):2640-2646.[55]辛兵,王敏.选择性环氧化酶-2抑制剂对卵巢癌细胞抑制作用的体内体外研究[J].现代肿瘤医学,2015,23(18): 2561-2565.[56]K E R S U A L N,G A R AM B O I S V,C HA R DÈS T,e t a l.T h eh u m a n M u l l e r i a n i n h i b i t i n g s u b s t a n c e t y p eⅡr e c e p t o r a si mm u n o t h e r a p y t a r g e t f o r o v a r i a n c a n c e r[J].m A b s, 2014,6(5):1314-1326.[57]K I M H S,S U N G Y J,P A I K S.C a n c e r c e l l l i n e p a n e l se m p o w e r g e n o m i c s-b a s e d d i s c o v e r y of p r e c i s i o n c a n c e rm e d i c i n e[J].Y o n s e i M e d J,2015,56(1):1186-1198.(收稿日期:2019-05-12修回日期:2019-08-19)㊃2703㊃国际检验医学杂志2019年12月第40卷第24期I n t J L a b M e d,D e c e m b e r2019,V o l.40,N o.24。
新高考英语题型精析精练与话题拓展：话题拓展07.植树造林与森林保护(解析版)
07.植树造林与森林保护养成良好的答题习惯，是决定高考英语成败的决定性因素之一。
做题前，要认真阅读题目要求、题干和选项，并对答案内容作出合理预测;答题时，切忌跟着感觉走，最好按照题目序号来做，不会的或存在疑问的，要做好标记，要善于发现，找到题目的题眼所在，规范答题，书写工整;答题完毕时，要认真检查，查漏补缺，纠正错误。
一、阅读理解1Recently, China has announced the list of the first five national parks. Each of them is divided into two parts — the core protection area and the general control area. In the core protection area, only research and surveillance (监视) in science are allowed. And the general control area is open to the public, allowing travel activities such as camping and hiking. In the future, national parks are expected to be natural classrooms. People can learn about different kinds of animals and plants through eco-friendly travel activities.Three River-Source National Park on the Qinghai-Tibet Plateau is the largest national park in China. Because it is home to the sources of the Yangtze, Yellow and Lancang rivers, people call it “China’s water tower”.Northeast China Tiger and Leopard National Park is in Heilongjiang and Jilin Provinces. It covers an area of 14,100 square kilometers. It is our country’s biggest and only place for wild Siberian tigers and Amur leopards to live in.Wuyi Mountain National Park in Fujian Province is a UNESCO natural and cultural heritage site. The forest makes up over 96 percent of the park. The park is the paradise of birds, kingdom of snakes and world of insects. You can also see the Danxia landform there.Giant Panda National Park connects panda habitats in Sichuan, Shaanxi and Gansu Provinces. Inside the park are more than 1,600 wild giant pandas. They make up over 70 percent of the pandas in China.Hainan Tropical Rain forest National Park is the largest tropical forest in China. There aremore than 400 kinds of plants that can only be found in Hainan.1. Which is called “China’s water tower”?A. Three River-Source National Park.B. Northeast China Tiger and Leopard National Park.C. Giant Panda National Park.D. Hainan Tropical Rainforest National Park.2. The underlined word “paradise” means ________.A. a perfect placeB. a happy feelingC. a good exampleD. a sweet smile3. Which of the following is not mentioned?A. Three River-Source National Park is the largest national park in China.B. Northeast China Tiger and Leopard National Park covers an area of 14,100 squarekilometers.C. There are more than 1,600 wild giant pandas in Giant Panda National Park.D. More than 400 kinds of plants and animals can only be found in Hainan.4. What might be the best title for the text?A. The beautiful scenes of natureB. The home of animals and plantsC. The introduction to the five national parksD. The relationships among the five nationalparks【答案】1. A 2. A 3. D 4. C【解析】这是一篇说明文，本文主要介绍了中国公布的首批五个国家公园的情况。
环磷酰胺辅助化疗对不同年龄乳腺癌患者卵巢功能的损伤情况及机制研究
・850・现代生物医学进展biomecLcnj Progress in Modern Biomedicine VoL21NO.5MAR.2021doi:10.13241/ki.pmb.2021.05.010环磷酰胺辅助化疗对不同年龄乳腺癌患者卵巢功能的损伤情况及机制研究*刘思旋李晓林陈璇马丽思翁怀玉宗祥云△(上海交通大学附属第六人民医院乳腺外科上海200233)摘要目的：以月经变化情况及血清激素水平为观察指标,探讨环磷酰胺辅助化疗方案对绝经前不同年龄段乳腺癌患者卵巢功能的损伤情况及可能的机制研究。
方法：收集2017年1月至2018年12月期间在我院接受环磷酰胺辅助化疗的绝经前乳腺癌患者77例患者年龄范围为26-48岁，并按年龄分为两组:M35岁组为16例；>35岁组为61例调查研究两组患者化疗前后的月经情况，以及血清激素水平(E、FSH、AMH)。
同时进行动物实验，采用单剂量环磷酰胺(200mg/kg)处理不同月龄组(3月龄和6月龄)雌性C57BL/6J小鼠，每组小鼠12例.在环磷酰胺处理前及处理后一周，检测小鼠血清AMH水平，计数小鼠卵巢卵泡数量结果：环磷酰胺化疗后，>35岁组与M35岁组间比较,>35岁组闭经率更高，复经率更低，血清AMH和E：水平下降幅度更大(P<0.05), FSH水平升高幅度也大于M 35岁组，但差异无明显统计学意义(P>0.05).不同年龄组内比较,W35岁组患者化疗后的血清AMH 和E?下降水平无明显差异(P>0.05)；血清FSH水平明显升高(M0.05),而>35岁组的患者化疗后，血清AMH和E：水平明显下降,FSH水平明显升高，差异均有统计学意义(片0.05)不同月龄组小鼠环磷酰胺处理前后，血清AMH水平明显下降，卵巢原始卵泡和生长卵泡数明显较少，差异均有统计学意义(P<0.05),且6月龄组小鼠下降程度更明显(P<0.05).结论：环磷酰胺辅助化疗方案明显损伤患者卵巢功能，且对年龄越大者损伤程度越严重，更易导致不可逆性卵巢损伤关键词:环磷酰胺；卵巢损伤；抗缪勒氏管激素中图分类号：R737.9;R730.53文献标识码：A文章编号：1673-6273(2021)05-850-07Study on the Damage and Mechanism of Cyclophosphamide-assistedChemotherapy on Ovarian Functionin Patients with Breast Cancer at Different Ages*LIU Si-xuan,LI Xiao-lin,CHEN Xuan,MA Li-si,WENG Huai-yu,ZONG Xiang-yurr' (Department of B reast Surgery,Shanghai Jiao Tong University Affiliated Sixth People's Hospital,Shanghai,200233,China) ABSTRACT Objective:To study the damage of cyclophosphamide adjuvant chemotherapy on ovarian function in different age groups of premenopausal breast cancer patients and the possible mechanism,the changes of menstruation and serum hormone level were observed.Methods:77premenopausal breast cancer patients who received cyclophosphamide adjuvant chemotherapy in our hospital from January2017to December2018were collected.The age range of the patients was26-48years old,and they were divided into two groups by age:16cases in the group of S35years old and61cases in the group of>35years old.To investigate the menstruation and serum hormone levels(E,FSH,AMH)of the two groups before and after chemotherapy.At the same time,a single dose of cyclophosphamide(200mg/kg)was used to treat female C57BL/6J mice of different age groups(3and6months),12mice in each group.Before and one week after cyclophosphamide treatment,the serum AMH level and the number of ovarian follicles were measured. Results:After cyclophosphamide adjuvant chemotherapy,the>35-year-old group had a higher amenorrhea rate and lower menstrual recovery rate than the S35-year-old group.The levels of serum AMH and E2decreased more significantly(P<0.05),and the increase of FSH levels was greater than S35year-old-group,but the difference was not statistically significant(P>0.05).Comparing within different age groups,there was no significant difference in the levels of serum AMH and E2after chemotherapy in patients S35years old(P> 0.05);but the levels of serum FSH significantly increased after chemotherapy(P<0.05).In>35-year-old group,serum levels of AMH and E2decreased significantly,and FSH levels increased significantly after chemotherapy.The differences were statistically significant(P<0.05).Before and after cyclophosphamide treatment in mice of different month-old groups,serum AMH levels decreased significantly,and the number of ovarian primordial follicles and growing follicles was significantly less.The differences were statistically significant(P<0.05),and the decrease was more significant in the6-month-old group(P<0.05).Conclusions:Cyclophosphamide adjuvant chemotherapy significantly damages the ovarian function of patients,and the older the patients are,the more serious the damage is,the more likely to *基金项目：上海市科学技术委员会西医引导项目(15411966500)作者简介:刘思旋(1994-),女,硕士研究生，主要研究方向：乳腺癌的诊疗，电话：怡217678801,E-mail:***************△通讯作者淙祥云(1974-),男，研究生导师,副教授，主要研究方向：乳腺癌的诊疗,E-mail:*****************(收稿日期：2020-07-23接受日期：2020-08-18)现代生物医学j o Progress in Modem Biomedicine VoL21NO.5MAR2021・851・lead to irreversible ovarian damage.Key words:Cyclophosphamide;Ovarian damage;Anti-Mullerian hormone(AMH)Chinese Library Classification(CLC):R737.9;R730.53Document code:AArticle ID:1673-6273(2021)05-850-07刖g近年来，由于乳腺癌诊疗手段的完善，乳腺癌的死亡率持续下降，早期5年生存率高达90%吓。
Thyroid Hormone Receptor beta (TR-beta) and Liver X Receptor(LXR)
Thyroid Hormone Receptor(TR)and Liver X Receptor (LXR)Regulate Carbohydrate-response Element-binding Protein(ChREBP)Expression in a Tissue-selective Manner*□S Received for publication,May19,2010,and in revised form,June24,2010Published,JBC Papers in Press,July8,2010,DOI10.1074/jbc.M110.146241Karine Gauthier‡1,2,Cyrielle Billon‡1,Marie Bissler‡,Michel Beylot§,Jean-Marc Lobaccaro¶,Jean-Marc Vanacker‡, and Jacques Samarut‡From the‡Institut de Ge´nomique Fonctionnelle de Lyon,Universite´de Lyon,Universite´Lyon1,CNRS,INRA,Ecole Normale Supe´rieure de Lyon,46alle´e d’Italie,69364Lyon,France,§Inserm ERI22/EA4173,Faculte´Rockefeller,Universite´Lyon1,69373Lyon,France,and the¶UMR,CNRS6247,Clermont Universite´,Centre de Recherche en Nutrition Humaine d’Auvergne, 63177Aubie`re Cedex,FranceThyroid hormone(TR)and liver X(LXR)receptors are tran-scription factors involved in lipogenesis.Both receptors rec-ognize the same consensus DNA-response element in vitro.It was previously shown that their signaling pathways interact in the control of cholesterol elimination in the liver.In the present study,carbohydrate-response element-binding pro-tein(ChREBP),a major transcription factor controlling the activation of glucose-induced lipogenesis in liver,is charac-terized as a direct target of thyroid hormones(TH)in liver and white adipose tissue(WAT),the two main lipogenic tis-sues in ing genetic and molecular approaches, ChREBP is shown to be specifically regulated by TRbut not by TR␣in vivo,even in WAT where both TR isoforms are expressed.However,this isotype specificity is not found in vitro.This TRspecific regulation correlates with the loss of TH-induced lipogenesis in TR؊/؊mice.Fasting/refeeding experiments show that TRis not required for the activation of ChREBP expression particularly marked in WAT following refeeding.However,TH can stimulate ChREBP expression in WAT even under fasting conditions,suggesting completely independent pathways.Because ChREBP has been described as an LXR target,the interaction of LXR and TRin ChREBP regulation was assayed both in vitro and in vivo.Each recep-tor recognizes a different response element on the ChREBP promoter,located only8bp apart.There is a cross-talk between LXR and TRsignaling on the ChREBP promoter in liver but not in WAT where LXR does not regulate ChREBP expression.The molecular basis for this cross-talk has been determined in in vitro systems.De novo lipogenesis allows the synthesis of new molecules of fatty acids from acetyl CoA.High glucose and insulin concen-trations induce this process,converting the excess energy into triglycerides,a more relevant molecule for storage purposes.In rodents,both liver and WAT3are efficient sites for lipogenesis. The synergic actions of insulin and glucose on the expression of lipogenic genes are mediated by key transcription factors. Insulin acts mainly through SREBP(sterol regulatory element-binding protein)-1c(1),whereas carbohydrate-response ele-ment-binding protein(ChREBP)is the master factor for glu-cose-induced lipogenesis(2).ChREBP physiological function has mainly been studied in the liver.ChREBPϪ/Ϫmice display a diminution in both basal and glucose-induced liver fatty acid synthesis due to the decreased expression of ChREBP glycolytic and lipogenic targets(3).Most interestingly,the ChREBPϪ/Ϫmutation protects Ob/Ob mice from obesity and reduces their plasma glucose level(4),suggesting that inhibition of ChREBP might be of pharmacological interest to treat the metabolic syndrome.ChREBP is expressed in many other tissues including WAT,where its possible lipo-genic role is presently unclear.ChREBP activity is mainly regulated by post-translational modifications that control its relocation to the nucleus and its DNA binding activity(5).When active,ChREBP turns on the expression of genes harboring a ChoRE(carbohydrate-re-sponse element)in their promoters.All the genes encoding the enzymes involved in lipogenesis(FAS,ACC,SCD1,L-PK, G6PD,ME,and Spot14)are direct ChREBP targets.During fast-ing,ChREBP is inactivated and located in the cytoplasm.In contrast ChREBP mRNA level varies in a narrow range.In liver, its level doubles when animals are switched from a fasted to a fed state(6).A similar up-regulation of its expression can be observed in mouse and human hepatocytes exposed to a high glucose concentration(7).In3T3-L1cells,insulin,glucose,and fatty acids regulate ChREBP expression(8).In contrast to liver, ChREBP mRNA is very efficiently induced(10-fold)following refeeding in WAT(6,8).The physiological consequence of this regulation in WAT remains unknown.*This work was supported by SIGNATOR Grant ANR-06-BLAN-0232-01,Cre-scendo Contract LSHM-CT-2005-018652,Cascade,Contract FOOD-CT-2004-506319,the Fondation pour la Recherche Me´dicale Grant INE2000-407031/1,and the Fondation BNP-Paribas.□S The on-line version of this article(available at )contains supplemental Table1and Figs.1–3.1Both authors contributed equally to this work.2To whom correspondence should be addressed:IGFL,UMR5242,ENS de Lyon,46alle´e d’Italie,69364Lyon cedex07,France.Tel.:33-0-472728616;Fax:33-0-472728080;E-mail:kgauthie@ens-lyon.fr.3The abbreviations used are:WAT,white adipose tissue;LXR,liver X receptor;RXR,retinoid X receptor;LXRE,LXR-response element;ChREBP,carbohy-drate-response element-binding protein;SREBP,sterol regulatory ele-ment-binding protein;TH,thyroid hormones;TR,thyroid hormonereceptor;TRE,TH-response elements;FAS,fatty acid synthase;PTU,propyl-thiouracil;qRT-PCR,quantitative RT-PCR;m,mouse;h,human.THE JOURNAL OF BIOLOGICAL CHEMISTRY VOL.285,NO.36,pp.28156–28163,September3,2010©2010by The American Society for Biochemistry and Molecular Biology,Inc.Printed in the U.S.A.at Karolinska institutet library, on February 16, Downloaded from/content/suppl/2010/07/08/M110.146241.DC1.html Supplemental Material can be found at:Thyroid hormones(TH)up-regulate lipogenesis in liver,but their roles in WAT are controversial(9–11).Their actions are mediated by the TR␣and TRnuclear receptors,which act as transcription factors by binding to specific TH-response ele-ments(TRE)as homodimers or heterodimers with the nuclear receptor RXR(12).Several genes involved in lipogenesis such as FAS,ACC,Spot14,or ME are positively regulated by TH in liver (13,14).TRE have been identified in some but not all of their promoters.The expression patterns of TR␣and TRare only partially overlapping(15).In liver,TRrepresents80%of the TH-bound TR(16),whereas in WAT,both receptors are highly expressed.The phenotyping of different TR KO mice sheds light on the role of each isotype in mediating TH signal(12,17). Importantly in the organs where they are co-expressed,their function is not necessarily redundant.Recently,two genes were described to be specifically regulated by either TR␣1or TR(18)in the outer hair of the developing cochlea,suggesting that each receptor might regulate its own set of targets in response to TH.The lipogenic effect of TH has been attributed to TRbecause in the liver,TH regulation of FAS,ACC,Spot14,and ME is lost in TRϪ/Ϫmice(13).However,because TR␣is weakly expressed in this tissue,liver might not be the most appropriate tissue to assay isotype specificity.The LXR nuclear receptors could be involved in the lipogenic action of TR.Dif-ferent levels of potential cross-talk between LXRs and TRhave indeed been described(19).For instance,LXR␣expres-sion has been previously described to be regulated by TH in mouse liver(20).At a functional level,LXRs and TRregulate a common set of events especially in the liver,where both recep-tors stimulate lipogenesis and cholesterol disposal.From a molecular point of view,these receptors can bind to identical (DR4)elements in vitro,although only one of these elements(in the cyp7a1gene promoter)has been characterized as a com-mon LXR-and TR-response element(21).Interestingly,LXRs were recently shown to directly control ChREBP expression by binding to a DR4element in its promoter(22).Another DR4 element located in the near vicinity was shown to mediate the positive effect of TH on ChREBP expression in the mouse liver (23).In this study,we show that TH directly activate ChREBP not only in liver(23)but also,to a higher extent,in WAT.In vivo, this effect is TR-,but not TR␣-,dependent,although both TR isoforms are strongly expressed in WAT,whereas in vitro,both isoforms can drive the expression of a reporter gene down-stream of the ChREBP promoter and bind to the same response element.Despite its capacity to up-regulate ChREBP expres-sion,TRis not required for ChREBP induction in response to the fasting/refeeding protocol.TRacts independently of LXR. Finally,although ligands for these receptors could co-regulate the ChREBP promoter in liver,different approaches point out to a mutually exclusive binding of LXR and TR to this promoter. EXPERIMENTAL PROCEDURESPlasmids—The expression plasmids were all pSG5-based vectors(mouse TR␣,rat TR1,mouse RXR␣,and mouse LXR␣).The different promoters were cloned in the pGL3basic vector and/or PGL4.70(hRluc)(Promega,Charbonnie`res, France).The3kbp upstream of the mouse ChREBP transcrip-tion start site were amplified by PCR using the primers ChREBPprom,cloned in pGL3basic/PGL4.70(pChREBP).The mutants(pM1,pM2,and pM1M2)were obtained using site-directed PCR mutagenesis(with M1and M2primer pairs).All plasmids were sequenced(Cogenics Genome Express,Meylan, France).Chemicals—Tri-iodothyronine(T3)and thyroxine(T4)were from Sigma-Aldrich(l’Isle D’Abeau,France),and the synthetic LXR ligand T0901317(T09)was from Cayman Chemical (Montigny le Bretonneux,France).Animals and Preparation of Tissue Samples—Knock-out mice were in a C57black6:129sv mixed background.TRϪ/Ϫ, TR␣0/0(17,24,25),LXR KO(26),and controls were fed ad libitum A04diet(SAFE,Augy,France),and housed under rec-ommended conditions.3–5-month-old male mice were used unless indicated otherwise.TH deficiency in adult animals was induced as described with a PTU-containing diet(Harlan Tek-lad TD95125,Madison,WI)and followed or not by TH(mix of T4and T3)injection(13).T09was given by oral gavage once a day for3days(10mg/kg of T09in100l of methyl cellulose 1%).Pax8Ϫ/Ϫmice,which are genetically hypothyroid,were previously described to die before weaning(27);however,some spontaneously survived.These rare survivors were used for experiments.For the fasting/refeeding protocol,mice fed a reg-ular chow diet were fasted for24h and either refed a70%high carbohydrate diet(Harlan Teklad TD98090)or kept on fasting for an additional16h.Tissues were dissected immediately after cervical dislocation and flash-frozen in liquid nitrogen.For WAT ex vivo culture,peritesticular fat pads were dissected and cultured non-dilacerated in10%charcoal-stripped fetal bovine serum(FBS),5ng/ml insulin complemented DMEM(Invitro-gen,Cergy-Pontoise,France)for24h before the addition of ligands.All animal experiments were performed under animal care procedures and conducted in accordance with the guide-lines set by the European Community Council Directives (86/609/EEC).RNA Extraction and Expression Analyses by Relative Quan-titative RT-PCR(qRT-PCR)—RNAs were extracted using TRIzol(Invitrogen).Total RNA was converted to cDNA using the SuperScript II retrotranscription kit(Invitrogen).qRT-PCR analyses were performed using the Quantitect SYBR Green PCR kit(Qiagen,Courtaboeuf,France)on a Stratagene machine MX3000pro(Stratagene,La Jolla,CA).Duplicates were run for each sample.The results were analyzed accord-ing to the⌬⌬CT method(28).36B4was always used as the reference gene,and the control group was either the non-treated cells or the WT non-injected animals unless otherwise indicated.Cell Culture and Transient Transfection Assays—HeLa (ATCC-CCL2)and3T3-L1(ATCC-CL-173)cells were main-tained in DMEM supplemented with10%FBS(Invitrogen).For 3T3-L1,cells were induced to differentiate using insulin-dexa-methasone-Rosiglitazone mix.To observe a better response to T3,cells were switched to DMEM medium supplemented with 10%charcoal-stripped FBS before the experiments.T3was used at10Ϫ8M,and T09was used at10Ϫ5M.Cells were har-vested24h(ChIP or WAT explants)or36h(transient trans-fection assay)after ligand exposure.For transient transfections,TRand LXR Regulate ChREBP in a Tissue-selective Mannerat Karolinska institutet library, on February 16, Downloaded fromHeLa cells were seeded in24-well plates and transfected with ExGen(Euromedex,Souffelweyersheim,France)following the manufacturer’s recommendations and0.5g of final DNA. pSG5was added as a carrier when needed.Transfection effi-ciency was normalized using-Gal activity brought by co-transfection of CMV-Gal vector.For each experiment,tripli-cates of each conditions were done,and each experiment was repeated at least three times,giving similar results.Only one experiment is shown;each point represents the average for the triplicate,and the error bar represents their S.D.Chromatin Immunoprecipitation Assays—The anti-TR␣an-tibody was raised against a C-terminal peptide and affinity-purified with the same peptide;the anti-TR(TR-J52)and con-trol IgG(normal mouse IgG)antibodies were purchased from Santa Cruz Biotechnology,and the anti-RNA polymerase II (CTD4H8)from Upstate Biotech Millipore.Cells were cross-linked with1%formaldehyde before lysis(in1%SDS,10m M EDTA,50m M Tris-HCL,pH8.1)and sonication(200–700bp DNA fragments).Lysates were diluted and precleared with her-ring sperm DNA(2g/ml),BSA(2g/ml),mouse IgG,and protein G-Sepharose(GE Healthcare,Saint-Cyr au Mont d’or, France).Lysates were incubated with the cognate specific anti-bodies or IgG and protein G-Sepharose.Beads were washed and eluted.Cross-link was reversed by overnight incubation at 65°C in the presence of RNase A and200m M NaCl.Samples were purified(Qiagen)and analyzed by quantitative PCR using the primer pairs NS1,NS2,and S1.EMSA—mTR␣1,mTR,mLXR␣,and mRXR␣were in vitro translated(T N T kit,Promega).The different single-strand oli-gonucleotides(forward)were[␥-32P]ATP-labeled with T4 polynucleotide kinase(Fermentas,Burlington,Ontario)before annealing with their unlabeled antisense(reverse).Probes were purified and counted.20,000cpm were used for each bindingreaction.Unlabeled specific and nonspecific competitor probes were included at the indicated molar excess.Hepatic Lipogenesis—Mice were given an i.p.injection of deuterated water(10ml/kg in0.9%NaCl isotonic water)fol-lowed by administration of drinking water enriched with deu-terated water(3%v/v)ad libitum for24h.Plasma was then collected for the measurement of deuterium enrichment in plasma water and in the palmitate of plasma triglycerides as described previously(29).These enrichments were then used to calculate the contribution,expressed as percentage,of hepatic lipogenesis to the plasma triglyceride pool(30). Statistics—For mice experiments,the data presented repre-sent the average values for the different animals(4or5)from the same genotype given the same treatment.The error bars represent S.E.Statistical relevance was determined using the one-variable analysis of variance method.All the primer sequences are listed in supplemental Table1.RESULTSChREBP Expression Is Regulated by TH in the Different Lipo-genic Tissues in a TR-dependent Manner—ChREBP expres-sion was recently shown to be regulated by TH in the liver of C57/BL6mice treated with PTU/Methimazole(23).Here the regulation of ChREBP was studied in the pax8(deprived of thyroid)mutant mice and Sv129mice treated with PTU(Fig.1A).In both models,TH injection induced ChREBP mRNA level in WAT and to a lesser extent in liver.Consistently,the expression of FAS(a target of both TRs and ChREBP)and L-PK (a ChREBP-only target gene)was also enhanced by TH,sug-gesting that ChREBP activity(and not only expression)is also up-modulated by TH.The TH-induced regulation of ChREBP was lost in TRϪ/Ϫbut not TR␣0/0mice,indicating that TRwas required at least in the two metabolic tissues studied(Fig. 1B)despite the strong expression of TR␣in WAT.The critical role of TRfor TH-induced hepatic lipogenesis was demon-strated in vivo using wild-type(WT)and TRϪ/ϪPTU-treated male mice.Although TH efficiently increased lipogenesis in WT(Fig.1C),the response was blunted in TRϪ/Ϫmice.WAT lipogenesis was not measured due to technical limitations. TH/TRand Nutritional Status,Two Independent Ways to Regulate ChREBP expression—To determine the involvement of TH signaling in the physiological regulation of ChREBP expression,RNA level was assessed in liver and WAT in response to a fasting/refeeding protocol in both WT and TRϪ/Ϫmice(Fig.2A).In agreement with published data, ChREBP RNA was found only up-regulated2-fold in the liver (6).In contrast,a dramatic increase of its expression was observed in WAT upon refeeding.This response was also observed in TRϪ/Ϫmice,indicating that TRis not required for this physiological process.We next determined whether FIGURE1.ChREBP expression and lipogenesis are regulated by thyroid hormones in a TR-dependent manner.3-month-old males either genetically rendered(pax8Ϫ/Ϫ)or chemically rendered(WT,TRϪ/Ϫ,TR␣0/0-PTU treated)hypothyroid were injected either by PBS(white bars)or by TH (black bars).In A and B,nϭ4for higher panel,nϭ5for lower panel(A)and (nϭ5)(B),mRNA encoding lipogenic enzymes were quantified by qRT-PCR. Veh,vehicle.In C,liver lipogenesis was measured as described under“Exper-imental Procedures”(nϭ5).Results are shown as induction as compared with the PTU-treated animals of a given genotype.Error bars represent S.E.Aster-isks and dollar signs indicate respectively statistical significance as compared with the PTU treatment of the same genotype and to the equivalent treat-ment in the WT group($or*,pՅ0.05,$$or**,pՅ0.005,$$$or***, pՅ0.0005).TRand LXR Regulate ChREBP in a Tissue-selective Mannerat Karolinska institutet library, on February 16, Downloaded fromChREBP expression could be TH-regulated under all nutri-tional conditions.ChREBP,as well as FAS and Spot14mRNAs, were induced by TH in the fasted(non-lipogenic)conditions in WAT(Fig.2B).In contrast,TH failed to significantly activate these genes when mice were refed.This might be due to an already high ChREBP expression under these conditions.In the liver,the extent of ChREBP mRNA regulation is much more limited,and in contrast to WAT,the nutrition signal is domi-nant,blocking a potential effect of TH on the three target genes in the fasted state.TRBinds to and Activates ChREBP Promoter via the Previ-ously Described LXRE2—The results presented above identified TH/TRas a new way to modulate ChREBP expression in vivo. The mechanisms responsible for this regulation were then investigated in vitro.In contrast to what was observed in vivo, TR,but also TR␣,when co-expressed with RXR␣,was able to activate the3.2-kbp ChREBP proximal promoter(Fig.3A)in the presence of TH(Fig.3B).LXR␣,previously described to activate this same portion of the promoter(22),was used as a positive control.Two DR4elements(LXRE1and LXRE2)were described in the mouse ChREBP promoter,with LXRE1being involved for LXR response(22)and LXRE2being necessary for TH response(23).These binding specificities were confirmed here by the EMSA data(Fig.3C).All three receptors bound to a 44-bp probe encompassing the two LXREs.However,LXR binding was competed only by an LXRE1WT but not mutated probe,whereas TR1or TR␣binding was only competed by an LXRE2WT but not mutated probe.The dependence on these sites for transcriptional responsiveness to either TR or LXR was less obvious in the transfection assay(Fig.3B).The double M1M2mutant still showed responsiveness to both compounds. This apparent discrepancy with EMSA results and published data for TR(23)is likely due to the inability of the four point mutations introduced in each promoter construct to efficiently prevent TR binding.For LXR,Cha and Repa(22)actually also observed a residual induction of similar pM1and pM1M2con-structs by the LXR agonist T09.This suggests either that besides LXRE1,some other region(s)of the promoter could mediate the response to LXR or that as for TR,the mutations introduced in LXRE1are not disruptive enough.ChIP experiments were performed to investigate the molec-ular mechanisms underlying the TR isoform specificity in the regulation of the endogenous ChREBP promoter.Differenti-ated3T3L1adipocytes in which ChREBP mRNA is also induced by TH were used.Similar to WAT,these cells express both TR␣and TR(31).Both TRs were detected on the region containing the LXREs but not on the upstream or downstream promoter regions.TR binding was independent of T3in agreement with the accepted model for TR action.In contrast,RNA polymerase II was enriched at the transcriptional start site only in the pres-ence of T3(Fig.3D).Altogether,these data clearly demonstrate that both TR␣and TRbind to the LXRE2in the ChREBP promoter and allow its induction in the presence of T3at least in a reporter system.Cross-talk between TRand LXR Signaling for the Regulation of ChREBP Expression—Published work described the LXR␣gene as a TH target in mouse liver(20).In the present study,no significant regulation of LXR␣expression by TH or T3was detected in the different models and experiments performed (Fig.4,A and D).Furthermore,TH was capable of activating the expression of ChREBP as well as other lipogenic genes in the liver of PTU-treated LXR KO mice(Fig.4A).The induction of ChREBP expression by TH is thus LXR-independent.TRand LXR activate the ChREBP promoter by respectively binding to LXRE2and LXRE1,two elements located in the close vicinity of each other.We thus assayed a potential functional interaction between the two signaling pathways.In liver but not in WAT, TH induction of ChREBP expression was significantly higher in LXR KO mice than in WT(4.5-fold versus2.9-fold,respectively, Fig.4A),suggesting that LXR might limit TRaccess to the promoter in WT liver.Such an increase is not observed for the regulation of other genes such as FAS,which is known to be regulated by both pathways.To document this interference for promoter binding,transfection experiments were performed in the presence of non-limiting amounts of RXR.Transfected alone,TRor LXR induced pChREBP activity in the presence of their cognate ligands(respectively,TH and T09).Remark-ably,co-transfection of both decreased the response to each ligand,LXR-dependent activity being more affected than TR (Fig.4B,left panel)by this inhibition.T09and T3displayed additive effects when both receptors were present.These obser-vations support the fact that concomitant binding of the two receptors to a single ChREBP promoter does not occur.This mutual inhibition was also observed to a lesser extent for both TR and LXR activities when increasing amounts of the other receptor were added.(Fig.4B,right panel).Finally direct evi-dences for a mutually exclusive binding were obtained by EMSA experiments.As shown previously in Fig.3,both recep-tors bind as RXR heterodimers to a44-mer probe containing the two WT LXREs.These two complexes migrated at the dif-FIGURE2.Independent regulation of ChREBP expression by TH/TRandnutritional status.WT and TRϪ/Ϫ3-month-old male mice were submittedto modification of nutritional and/or TH status.In A,mice were either fed aregular chow diet(CF)or starved for24h and then refed(R)or not(F).(nϭ5).In B,mice were starved for24h.One group was kept on fasting(F),and theother one was refed(R)for an additional16h.Half of the animals per groupwere injected by TH twice,once before the fast and then before the refeeding(nϭ5).Expression of lipogenic genes was measured by qRT-PCR.Asterisksindicate statistical significance(*,pՅ0.05,**,pՅ0.005,***,pՅ0.0005)ascompared with the CF group of the same genotype in A,to the F/V group in B).Dollar signs indicate statistical significance between F and RF groups in A andbridged groups in B($,pՅ0.05,$$,pՅ0.005,$$$,pՅ0.0005).In A,the valuefor the relative expression has been fixed to one in each genotype for the CFgroup.Error bars represent S.E.TRand LXR Regulate ChREBP in a Tissue-selective Mannerat Karolinska institutet library, on February 16, Downloaded fromferent sizes indicated on the figure.The LXR/RXR complex bound to the WT probe was gradually displaced by an increas-ing amount of TR /RXR,which noticeably failed to bind theprobe even at the highest amount added.We also observed that a TR /RXR complex was displaced by the addition of LXR/RXR.However,in both cases,the newly added complex was perfectly able to bind in a dose-dependent manner if the probe used contained a mutated version of the LXRE required for the fixation of the initially present receptor (M1for LXR and M2for TR).Altogether,these data strongly suggest that despite using two different LXREs,in this in vitro setting,concomitant binding of LXR/RXR and TR/RXR to the ChREBP promoter fragment is prevented.As a complementary way to analyze the interference between LXR and TR signaling pathways,mice or WAT explants were treated with different combinations of LXR and TR ligands (Fig.4D ).The efficiency of the different treatments was val-idated by measuring the expression levels of known LXR or TR targets in the two considered systems.In WAT explants,all genes behaved as expected with strong induction of ABCA1,SREBP1c,and ApoE by T09,whereas ChREBP and FAS were stimulated by T3.Surprisingly,in these same samples,LXR ligand failed to induce ChREBP expres-sion.Co-treatment with both ligands did not yield any additional effect as compared with treatment with individual ligand for any of the target tested.This suggests that TR and LXR mainly possess a non-over-lapping set of targets in WAT.In liver,treatments were also efficient,with an increase of both ChREBP and FAS by TR and LXR ligand alone.In this condition,ChREBP induction by T09does not reach statistical significance,but lack of strong induction has already been described by others (32).Co-treatment with T09and TH led to a significant increase inChREBP as well as FAS liver expres-sion as compared with TH treat-ment alone.This suggests that the two signals can be additive in this organ.For SREBP1-c and ABCA1,the situation is more complex.In PTU-treated mice,no activation was detected by T09alone,and TH repressed expression of both genes.Nonetheless,T09strongly in-creased their expression in TH-treated animals.Altogether,these data demonstrate that TR and LXR are both active in the two lipo-genic tissues,WAT and liver,although their target genes are different in vivo and depend on the tissue considered.DISCUSSION In this report,we show that in mice,ChREBP is a new direct TH target not only in liver,which is in agreement with recently published data (23),but also to a much higher extent in WAT.Careful dissection of the molecular mechanism of ChREBP reg-ulation allowed us to demonstrate that TR ,but not TR ␣,is required for this activity in vivo and interferes with LXR signaling.TH Stimulate ChREBP Expression in a TR -dependent Man-ner in Liver and WAT —TH have been long known to regulate energy metabolism and lipogenesis in the liver (9–11),yet their lipogenic effect in other tissues such as WAT was still contro-versial.Measurement of in vivo hepatic lipogenesis demon-strates that TH induction of this process is TR -mediated because it was abrogated in TR Ϫ/Ϫ.Notably,this regulation by TR correlates with its ability to up-regulate ChREBP expres-FIGURE 3.TR ␣and TR bind to and activate ChREBP promoter via the previously described LXRE2.A ,scheme of the different versions of the ChREBP promoter cloned upstream of a luciferase reporter.LXRE1and LXRE2are pictured as black ovals or white when mutated.The top arrow indicates the transcription start site.The arrow pairs below the promoter indicate the localization of the primers used for ChIP analyzes:white for NS2,black for S1,and gray for NS1.The regions amplified by these three pairs are respectively the promoterportion Ϫ4100/Ϫ3900,Ϫ2558/Ϫ2384,and Ϫ203/ϩ4.B ,the indicated promoters were transfected with TR ␣,TR ,or LXR ␣together with an RXR ␣encoding plasmid and treated with vehicle (veh ,white ),T3(light gray ),or T09(dark gray ).The relative luciferase activity measured is reported as arbitrary units (RAU ).C ,EMSA wereperformed using a 44-bp-long probe from the ChREBP promoter (WT probe)containing the area with the twoLXREs to detect TR /RXR ␣or TR ␣/RXR ␣binding.LXR ␣/RXR ␣has been included as a control.The asterisks indicate the specific petition with 100-fold excess of cold smaller fragments containing only one of the two LXREs,either WT (LXRE1or LXRE2)or mutated (LXRE1mut or LXRE2mut),was used to assess thespecificity of the binding.mut ,mutation.D ,ChIP experiments were performed on differentiated 3T3-L1,treated (light gray )or not (white )with T3.On the left are results obtained with anti-TR ␣(TR ␣),anti-TR (TR ),or mouse IgG (IgG ).On the right are results obtained with anti-RNA polymerase II (RPII )and mouse IgG (IgG ).The specificity of the antibodies (Ab )used was verified on transfected HeLa cells (supplemental Fig.1).The samelysates were used for all conditions,and each precipitation was done in replicates.The results shown are an average of these duplicates.Each experiment has been repeated at least twice.The primer pairs used for detection are indicated under the arrows .Error bars represent S.D.TR and LXR Regulate ChREBP in a Tissue-selective Mannerat Karolinska institutet library, on February 16, 2011 Downloaded from。
MHT治疗围绝经期综合征患者效果评价
*临床药物研究*MHT治疗围绝经期综合征患者效果评价分析刘端芳新泰市妇幼保健计划生育服务中心妇女保健科，山东新泰271200摘要目的深入分析绝经雌激素替代疗法（menopause hormone therapy，MHT）治疗围绝经经期综合征患者的应用效果。
方法选取2019年11月—2020年11月新泰市妇幼保健计划生育服务中心90例围绝经期综合征患者，遵循随机数表法均分成两组，其中45例接受常规药物治疗的患者设为对照组，另外45例接受MHT的患者设为观察组，对比两组围绝经期综合征患者的临床疗效、卵巢功能指标、不良反应发生率。
结果治疗后，观察组治疗有效率93.3%高于对照组治疗有效率77.8%，差异有统计学意义（χ2=4.406，P<0.05）；观察组FSH、E2等激素水平改善情况明显，与对照组相比改善情况更好，差异有统计学意义（P< 0.05）；观察组不良反应发生率比对照组低，但差异无统计学意义（P>0.05）。
结论围绝经期综合征患者接受MHT治疗后，患者卵巢功能明显改善，调节雌激素水平的同时降低了各类不良反应的发生，治疗有效性、安全性受到了许多人认可，值得临床推广。
关键词围绝经期综合征；雌激素替代疗法；治疗效果；卵巢功能中图分类号R711文献标志码A doi10.11966/j.issn.2095-994X.2022.08.09.46Effect of MHT in the Treatment of Patients with Perimenopausal SyndromeLIU DuanfangDepartment of Women´s Health,Maternal and Child Health and Family Planning Service Center,Xintai,Shandong Province,271200ChinaAbstract Objective To analyze the application effect of estrogen replacement therapy(MHT)in the treatment of patients with perimenopausal syndrome.Methods From November2019to November2020,90patients with perimenopausal menstrual syndrome in Xintai Maternal and Child Health and Family Planning Service Center were selected.Divided them into two groups according to the random number table method. Among them,45patients who received conventional drug treatment were set as the control group,and the other45patients who received MHT were set as the observation group.The clinical efficacy,ovarian function indexes and incidence of adverse reactions were compared between the two groups of patients with perimenopausal syndrome.Results After treatment,the treatment effective rate in the observation group was 93.3%higher than that in the control group,which was77.8%,and the difference was statistically significant(χ2=4.406,P<0.05);the im‐provement of FSH,E2and other hormone levels in the observation group was significantly better than that in the control group,and the differ‐ence was statistically significant(P<0.05);compared with the control group,the incidence rate of adverse reactions of the observation group was significantly lower,and the difference was not statistically significant(P>0.05).Conclusion Ovarian function was significantly improved in patients with perimenopausal syndrome who received MHT estrogen replacement therapy.While regulating the level of estrogen,it reduces the occurrence of various adverse reactions.The efficacy and safety of the treatment have been recognized by many people,and worthy of clinical promotion.Key words Perimenopausal menstrual syndrome;Estrogen replacement therapy;Therapeutic effect;Ovarian function 围绝经期综合征在临床上也叫做更年期综合征，典型表现为绝经前后的女性会出现较大的性激素波动，收稿日期：2022-07-05；修回日期：2022-07-28作者简介：刘端芳（1976-），女，本科，副主任医师，研究方向为女性全生命周期保健、生殖内分泌治疗、计划生育技术。
儿科规培题库名词解释07
1-脱氧-8-右旋精氨酸血管加压素，为人工合成制剂。抗利尿作用强，血管收缩作用弱，药效可达12小时以上。
44.外周性性早熟(peripheralprecocious puberty)
对非受控下生脑—垂体—性腺功能所引起的性早熟，有第二性征发育，有性激素水平升高，但下丘脑—垂体—性腺轴不成熟，无性腺的发育。
1.酮症性低血糖(ketotic hypoglycemia)
指低血糖发生的同时伴有尿酮体阳性，说明机体在反调节激素刺激下已动员脂肪组织释放出脂肪酸供能，多数年长儿酮症性低血糖是特发性的。
2.Ⅱ型糖尿病(noninsulin-dependent diabetes mellitus NIDDM)
对β细胞分泌胰岛素不足或(和)靶细胞对胰岛素不敏感所致的糖尿病，亦称非胰岛素依赖型糖尿病。
45.尿崩症(DiabetesInsipidus，DI)
对由于患儿完全或部分丧失尿浓缩功能所致，主要表现为多饮、多尿和排出稀释性尿。其中较多见的是由于血管升压素分泌或释放不足引起者，称中枢性尿崩症。
46.射血分数(ejectionfraction EF)
射血分数是指心每搏输出量占心室舒张末期容积的百分比。
位于细胞表面或细胞内，结合特异激素并引发细胞响应的蛋白质。
24.部分性性早熟
对指单纯乳房发育、单纯性阴毛发育、单纯性早初潮。
25.自分泌(anticrime)
激素可以与分泌它的自身细胞发生作用，是一种分泌细胞的自身调节作用。
26.家族性矮身材
父母身高矮，小儿身高常在第三百分位数左右，但其年生长速率大于4cm/年，骨龄和年龄相称，智能和性发育正常。
女孩在8岁、男孩在9岁以前呈现性发育征象，既为性早熟；性征与起真实性别一致者为同性性早熟，否则为异性性早熟。
亮丙瑞林对ER阳性绝经前乳腺癌化疗患者卵巢功能及骨密度的影响
亮丙瑞林对ER阳性绝经前乳腺癌化疗患者卵巢功能及骨密度的影响作者：罗军彭积院潘铃娟来源：《中国医学创新》2024年第07期【摘要】目的：探討亮丙瑞林对雌激素受体（ER）阳性绝经前乳腺癌化疗患者卵巢功能及骨密度（BMD）的影响。
方法：选取2020年1月—2022年8月丰城市人民医院收治的82例ER阳性绝经前乳腺癌患者的病历资料进行回顾性分析，根据治疗方式分为化疗组（n=41）和亮丙瑞林组（n=41）。
化疗组患者予以AC-T辅助化疗，亮丙瑞林组在化疗组的基础上联合亮丙瑞林治疗。
比较两组患者疗效、卵巢功能、BMD及月经情况。
结果：亮丙瑞林组的总有效率高于化疗组（P<0.05）。
治疗后，两组血清雌二醇（E2）水平均明显降低，血清促黄体生成素（LH）、卵泡刺激素（FSH）水平均明显升高（P<0.05）；亮丙瑞林组患者的血清LH、FSH水平均明显高于化疗组，血清E2水平明显低于化疗组（P<0.05）。
治疗后，两组左髋部、腰椎的BMD水平均明显降低（P<0.05）；两组比较差异均无统计学意义（P>0.05）。
亮丙瑞林组的闭经时间、月经恢复正常时间均较化疗组短，月经复潮率较化疗组高（P<0.05）。
结论：亮丙瑞林可保护ER阳性绝经前乳腺癌化疗患者卵巢功能，可有效提高疗效，但可能会导致患者BMD下降，需要在治疗期间注意监测BMD变化，降低骨质疏松发生风险。
【关键词】雌激素受体阳性绝经前乳腺癌化疗亮丙瑞林卵巢功能骨密度Effects of Leuprorelin on Ovarian Function and Bone Mineral Density in ER Positive Premenopausal Breast Cancer Patients Undergoing Chemotherapy/LUO Jun， PENG Jiyuan， PAN Lingjuan. //Medical Innovation of China， 2024， 21（07）： -126[Abstract] Objective： To investigate the effect of Leuprorelin on ovarian function and bone mineral density （BMD） of estrogen receptor （ER） positive premenopausal breast cancer patients undergoing chemotherapy. Method： The medical records of 82 ER positive premenopausal breast cancer patients admitted to Fengcheng People's Hospital from January 2020 to August 2022 were retrospectively analyzed， and they were divided into chemotherapy group （n=41） and Leuprorelin group （n=41） according to treatment methods. Patients in chemotherapy group were treated with AC-T adjuvant chemotherapy， and Leuprorelin group was treated with Leuprorelin on the basis of chemotherapy group. The curative effect， ovarian function， BMD and menstruation were compared between the two groups. Result： The total effective rate of Leuprorelin group was higher than that of chemotherapy group （P<0.05）. After treatment， serum estradiol （E2） levels in both groups were significantly decreased， and serum luteinizing hormone （LH） and follicle stimulating hormone （FSH） levels were significantly increased （P<0.05）; serum LH and FSH levels in Leuprorelin group were significantly higher than those in chemotherapy group， and serum E2 level was significantly lower than that in chemotherapy group （P<0.05）. After treatment，BMD levels of left hip and lumbar vertebrae were significantly decreased in both groups （P<0.05）; there were no significant differences between the two groups （P>0.05）. The amenorrhea time and menstrual normalcy time of Leuprorelin group were shorter than those of chemotherapy group， and the rate of menstrual rehydration was higher than that of chemotherapy group （P<0.05）. Conclusion： Leuprorelin can protect the ovarian function in patients with ER positive premenopausal breast cancer undergoing chemotherapy， which can effectively improve the therapeutic effect， but may lead to the decrease of BMD in patients， it is necessary to pay attention to the change of BMD during treatment to reduce the risk of osteoporosis.[Key words] Estrogen receptor positive Premenopausal breast cancer Chemotherapy Leuprorelin Ovarian function Bone minernal densityFirst-author's address： General Surgery Department Ⅱ， Fengcheng People's Hospital，Fengcheng 331100， Chinadoi：10.3969/j.issn.1674-4985.2024.07.029乳腺癌是常见的妇科恶性疾病，具有较高的发病率及死亡率，如不及时治疗，将严重威胁女性的生命健康，乳腺癌的常规治疗方式是以手术切除辅助化疗为主[1]。
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G蛋白受体介导的信号转导模式图
胞 外 细胞膜 胞 内
受体
第一信使
偶联蛋白
效 应 酶
第二信使
生化级 联反应
功能性 应答
第二信使
小分子细胞内信使的特点： ①在完整细胞中，该分子的浓度或分布在细胞外
信号的作用下发生迅速改变 ②该分子类似物可模拟细胞外信号的作用 ③阻断该分子的变化可阻断细胞对外源信号的反 应 ④作为别构效应剂在细胞内有特定的靶蛋白分子
这些脂类衍生物都是由体内磷脂代谢产生的。
1. G蛋白耦联受体途径—— GPCR
1. G蛋白耦联受体途径—— G蛋白, GTP-结合蛋白,
GTP binding protein Protein ①.三聚体G蛋白,与膜受体偶联 ②.结构：αβγ三种亚基 固定于细胞膜内侧 ③.特性：具GTP酶的活性
GTP
(一) 激素的受体 Receptor 指靶细胞上能识别并专一性结合某种 激素 , 继而引起生物效应的功能蛋白质 , 是细胞接受信息的装置。
1. 受体的分类(根据在细胞中的位置)
(1) 细胞膜受体 (2) 细胞内受体
胞浆受体：存在于胞浆中的可溶性特殊蛋白质, 与激素结合形成激素 -胞浆受体复合物,使激素转 移入核内发挥作用。
酶偶联受体种类繁多，但是以具有PTK活性和与 PTK偶联的受体居多。
2. 酶偶联受体途径
①结合配体后受体形成二聚体或寡聚体； ②第一个蛋白激酶被激活。对于具有蛋白激酶活性的受体 来说，此步骤是激活受体胞内结构域的蛋白激酶活性； 对于没有蛋白激酶活性的受体来说，此步骤是受体通过 蛋白质-蛋白质相互作用激活与它紧密偶联的蛋白激酶； ③通过蛋白质-蛋白质相互作用或蛋白激酶的磷酸化修饰 激活下游信号转导分子，通常是继续活化下游的一些蛋 白激酶； ④蛋白激酶通过磷酸化修饰激活代谢途径中的 关键酶、
P
P
RAS
SOS
GTP
P
RAS-RAF-MEK-MAPK
SOS • 180 kDa guanine nucleotide exchange factor for RAS; stimulate the formation of active GTP-bound RAS.
• SOS belongs to the DBL
P
P
RAS
★ ★ ★ ★ ★
★ ★ ★
★ ★
三十九肽 糖蛋白 糖蛋白 糖蛋白 十八肽 蛋白质 蛋白质 胺类 胺类 三十二肽 蛋白质 蛋白质 类固醇 类固醇 胺类 胺类
睾丸：间质细胞 支持细胞 卵巢、胎盘
消化道、脑
心房
睾 酮 抑制素（卵巢也可产生） 雌二醇 雌三醇 孕酮 人绒毛膜促性腺激素 促胃液素(胃泌素) 缩胆囊素 促胰液素 (胰泌素) 心房钠尿肽
核受体: 存在于核内,能与激素结合, 对细胞核内 转录过程起调节作用的蛋白质。
激素作用机制
蛋白质多肽类 氨基酸衍生物类 （水溶性激素） 类固醇类激素 甲状腺激素 视黄酸 （脂溶性激素）
1
2
细胞膜受体按作用模式可分为
G蛋白耦联受体(G-Protein Coupled Receptor)
酶偶联受体 受体酪氨酸蛋白激酶 受体Ser/Thr蛋白激酶 其他激酶系统
(二) 细胞膜受体的作用机制 第二信使学说 second messenger theory
激素 ＋
细胞膜受体
激活膜内效应器酶
细胞内产生第二信使
细胞发生生物学效应
激活PK
底物磷酸化
信号接收装置: 入室线座(电信号) -膜受体 信号转导装置: 座机 -转导蛋白 信号传出装置: 听筒(声音) -第二信使
肽 肽 肽 多巴胺(?) 九肽 九肽
促黑（素细胞）激素释放抑制因子 MIF
腺垂体
促肾上腺皮质激素 ACTH 促甲状腺激素 TSH 促卵泡激素 FSH 黄体生成素 LH 促黑（素细胞）激素 β─MSH 催乳素 PRL 生长素 GH 甲状腺 甲状腺素（四碘甲腺原氨酸） T4 三碘甲腺原氨酸 T3 甲状腺C细胞 降钙素 CT 甲状旁腺 甲状旁腺激素 PTH 胰岛 胰岛素 肾上腺：皮质 糖皮质激素（如皮质醇） 盐皮质激素（如醛固酮） 髓质 肾上腺素 E 去甲肾上腺素 NE
如一些离子通道也可以直接受cAMP或cGMP的别构调节。
蛋白激酶A是cAMP的靶分子
第二信使
脂类也可作为胞内第二信使
二脂酰甘油（diacylglycerol，DAG） 花生四烯酸（arachidonic acid，AA） 磷脂酸（phosphatidic acid， PA） 溶血磷脂酸（lysophosphatidic acid，LPA） 4-磷酸磷脂酰肌醇（PI-4-phosphate，PIP） 磷脂酰肌醇-4,5-二磷酸(phosphatidylinositol -4,5diphosphate，PIP2) 肌醇-1,4,5-三磷酸（Inositol-1,4,5-triphosphate，IP3）
SOS
GDP
P
P
family of oncogenes (14 members). Most of these activate the RHO family of GTP-binding proteins involved in cytoskeletal reorganization. SOS is one of the few known member of the family for RAS.
二、激素的分类
按其化学结构可ห้องสมุดไป่ตู้为 蛋白质多肽类
(一)含氮激素
氨基酸衍生物(胺类) (二) 类固醇 (甾体) 激素 (三) 脂肪酸衍生物
激素的传递方式
Telecrine Paracrine Autocrine Neurocrine
主
主要来源
下丘脑
要
激
激素
素
及
其
化
TRH
性
质
化学性质
三肽
英文缩写
GnRH
一、激素作用的一般特性
(一) 作用特异性：对所作用的器官、组织、细胞 具有选择性(故称靶*****)并选择性地与其特异 性受体结合 (二) 作用微量性： 0.1gCRH→ 1 gACTH→ 40 gcortisol (三) 可调控性：合成、分泌 (四) 信息传递作用: 仅起信使作用 (五) 作用“快”"慢" (六) 脱敏作用 (七) 激素之间的相互作用
• The SH3 of GRB2 binds constitutively to the proline-rich sequence in the C-terminus of SOS (a guanine nucleotide exchange factor for RAS).
P
P
SOS
P P
GRB2
GDP
cAMP cAMP
④.种类/功能: Gs: Stimulatory G protein Gi: inhibitory G protein Gq: 活化磷脂酶C-β
1. G蛋白耦联受体途径—— G Protein
Gs G蛋白 Gi
cAMP cAMP
当: Hs ＋Rs
Hi ＋ Ri
Gs
Gi
cAMP
GRB2
RAS-RAF-MEK-MAPK
P • RAS becomes activated by exchanging GDP for GTP
P
RAS
SOS
GTP GDP
P
P
GRB2
RAS-RAF-MEK-MAPK
RAS • 21 kDa GTPase protein.
• A special case of G protein monomer • GTP-bound form: active GDP-bound form: inactive. • originally identified as oncogenes in several retroviruses, and are activated in a variety of human tumors.
RAS-RAF-MEK-MAPK
GRB2 • 24 kDa adaptor molecule. • Only contains an SH2 domain between two SH3 domains
P
P
SOS
P P
GRB2
RAS-RAF-MEK-MAPK
GRB2 • The SH2 of GRB2 binds to phosphotyrosine in a number of receptor PTK: for EGF, PDGF, colonystimulating factor-1 (CSF1), stem cell factor (SCF);
cAMP
1. G蛋白耦联受体途径 GPCR (1) AC-cAMP-PKA途径
胰高血糖素受体通过AC-cAMP-PKA通路转导信号
(2) PLC - IP3/DG - CaM/PKC途径
2. 酶偶联受体途径
酶偶联受体指那些自身具有酶活性，或者自身没 有酶活性，但与酶分子结合存在的一类受体。 这些受体大多为只有1个跨膜区段的糖蛋白，亦 称为单跨膜受体。
GHRH
促甲状腺激素释放激素
促性腺激素释放激素
生长素释放激素
十肽
十四肽 四十四肽
生长素释放抑制激素（生长抑素） GHRIH
促肾上腺皮质激素释放激素
促黑（素细胞）激素释放因子 催乳素释放因子 催乳素释放抑制因子 血管升压素（抗利尿激素） 催产素
CRH
MRF PRF PIF VP (ADH) OXT
四十一肽
第七章 激素及其作用机制 hormone