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研究生:楊東川
研究生(外文):Yang, Tung-Chuan
論文名稱:控制性超排卵刺激誘發一氧化氮變化之研究
論文名稱(外文):Studies on the regulation of nitric oxide induced by controlled ovarian hyperstimulation
指導教授:蔡鴻德蔡鴻德引用關係
指導教授(外文):Tsai, Horng-Der
學位類別:碩士
校院名稱:中國醫藥學院
系所名稱:醫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2000
畢業學年度:88
語文別:中文
論文頁數:84
中文關鍵詞:體外受精胚胎移殖控制性超排卵一氧化氮內皮性一氧化氮合成■免疫組織化學西方墨點分析法
外文關鍵詞:IVF-ETcontrolled ovarian hyperstimulationNOeNOSImmunohistochemistryWestern blot analysis
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控制性超排卵刺激誘發一氧化氮變化之研究
背景利用外源性促性腺激素(gonadotropins)刺激卵巢產生超排卵作用(superovulation),來增進不孕夫婦的懷孕率,已是不孕症的治療常規之一。一氧化氮(nitric oxide, NO)最初由血管內皮細胞發現,是眾所周知的血管鬆弛因子;而一氧化氮也具有調節卵巢組織內卵泡成長、排卵以及性類固醇生成等生理功能。本論文將評估控制性超排卵刺激(controlled ovarian hyper-stimulation, COH)對於體外受精胚胎移植(in vitro fertilization and embryo transfer, IVF-ET)之婦女血液一氧化氮濃度之影響;並分析控制性超排卵刺激對於雌性小白鼠卵巢細胞內一氧化氮合成■(nitric oxide synthase, NOS)的表現。
實驗方法(1)雌性小白鼠動物實驗-以六至八週大雌性小白鼠,分成控制組、超排卵刺激組(Pregnant mare’s serum gonadotropin, PMSG 及 human chorionic gonadotropin, hCG)和超排卵刺激加NOS抑制劑(L-NG-nitro-D-arginine methyle ester, L-NAME)組共三組。以免疫組織化學法(Immuno-histochemical,IHC)和西方墨點分析法(Western blot analysis),來觀察卵巢組織各種細胞中內皮性一氧化氮合成■之濃度、時程變化(time-course changes)及特定性分佈(cell-specific distribution)。(2)生育期婦女超排卵刺激:實驗分為兩組;一組是正常月經週期的婦女,一組是接受試管嬰兒治療之不孕症婦女。共收集18名生育期婦女血液樣本。本研究將測量受試者血液中一氧化氮與動情素、黃體素濃度之相對變化;以探討控制性超排卵刺激下婦女,隨月經週期其體內一氧化氮濃度以及荷爾蒙(動情素及黃體素)變化之關聯性。
結果本論文發現雌性小白鼠卵巢組織內的膜細胞層(thecal cell layer)、基質細胞(stromal cell)、卵細胞的表面以及整個黃體內,皆可見明顯呈褐色的內皮性一氧化氮合成■(endothelial NOS, eNOS),且具有明顯的特定細胞分佈性質。而給于超排卵加NOS抑制劑的雌性小白鼠,卵巢組織僅有少數可見卵泡有黃體化,雖然褐色的eNOS表現依然存在。當以PMSG刺激卵泡成長後,可見eNOS表現隨著時間而逐漸增強;再以hCG觸發排卵後,eNOS亦隨著時間持續增加;而在黃體形成後達到高原期,呈現出明顯的時程變化。
本論文亦發現正常月經週期的婦女和進行控制性超排卵刺激之不孕症婦女,其血液中一氧化氮最高濃度是出現在月經週期之中週期(midcycle);此時血中動情素濃度也是最高。
結論本實驗證實在小白鼠的卵巢膜細胞層和基質細胞以及卵細胞的表面,皆可見到褐色的eNOS表現;以PMSG刺激後則會隨時程演變而逐漸顯現在壁顆粒細胞,以hCG取代LH surge觸發排卵後則逐漸顯現在黃體內,且呈現明顯的細胞特定性分佈形態。分析eNOS不同時間點的相對含量時,則呈現出明顯的時程變化,先以PMSG刺激可見eNOS表現逐漸增強,再以hCG觸發排卵後也持續增加,而在黃體形成後達到高原期。eNOS是組成性合成,在早期的顆粒細胞並無表現;至於,促性腺激素藉由何種機轉誘發晚期顆粒細胞合成eNOS?目前仍了解很少。
人類血中動情素和一氧化氮含量同時在濾泡晚期達到高峰,推論應與全身性血管擴張有關,以利於排卵和子宮內膜的生長。這些報告與發現可以支持動情素會調控一氧化氮的產生和釋出;但是中分泌期黃體素對於一氧化氮的產生和釋出可能有相反的作用,而這一推論未來須要更多的研究來加以證實。
Studies on the Regulation of Nitric Oxide Induced by Controlled Ovarian Hyperstimulation
Background
Controlled ovarian hyperstimulation with exogenous gonadotropins has become a routine part of infertility therapy. The improving uterine arterial blood flow and endometrial development as well as the large number of oocytes that can be obtained greatly increase the overall likelihood for pregnancy for infertile couples. NO, one of the most potent vasodilators, synthesized from the amino acid L-arginine, is now a well-established EDRF. NOS expression and NO generation have been recently reported in ovaries of rats, guinea pigs and humans. NO is also known to play several roles in folliculogenesis, ovulation and sex steroidogenesis.
The purpose of the present study was to localize eNOS protein in the mouse ovary and to test their hormonal regulation induced by gonadotropins. Moreover, using follicular development phase of menstrual cycle in normal women and patients undergoing IVF-ET treatment, we investigated whether ovarian sex steroids modulate NO production.
Methods
Animal Study:
Animals purchased from the supplier were given a few days to adjust to the light-dark cycle before the administration of the PMS. Mice were maintained on a 12 h light:12 h dark cycle and food and water ad libitum.
A total of 36-48 female mice were divided into 3 groups. Of these, all were given PMS on day 1.
(i)Control mice group.
(ii)Superovulation group.
(iii)Superovulation plus NOS inhibition group: Of these mice on PMS, 12-16 were given the NOS inhibitor, L-NAME (50 mg/mouse/24 h) as i.p. injections on days 1-3 of super-ovulation program.
Animals were chosen at random for tissue collection and future use for immunohistochemical studies as well as Western blot analysis. Three animals per moment observed were used to obtained tissues.
Study on women in reproductive age:
Two groups of women within reproductive age, 6 normally cyclic women as control group, another 12 women in stimulated cycle, were enrolled. Circulating level of NO and sex steroids (E2, P4) were checked at specific periods on cycle day 3, mid-cycle or the day just prior to hCG injection, and current mid-luteal phase. All blood samples were collected after overnight fasting and centrifuged immediately in a refrigerated centrifuge. The samples were frozen at —700C until analysis. The changes of circulating levels of NO and sex steroids during normal cycle or COH regimen were measured and discussed for study.
Results
Animal Study:
Using polyclonal rabbit antisera and immunohistochemical staining, the cell-specific expression of eNOS was demonstrated. Ovaries obtained from each group of mice contained follicles in early antral stages with multiple layers of granulosa cells surrounding oocytes. Brownish immunostaining indicated the presence of eNOS in the theca cell layer and stroma cells and on the surface of oocytes No specific staining was found within the granulosa cell layer in control mice. Many large antral follicles or corpus luteum were found after superovulation treatment.. At ovulation, no further changes in expression of eNOS were detected, but after ovulation and luteinization, enhanced eNOS staining was observed within the corpus luteum. In superovulation plus NOS inhibition group, brownish immunostaining remain, but decreased number of corpus luteum was present.
Using polyclonal rabbit antisera and Western blot analysis, quantitative data are consistent with the changes in eNOS expression observed immunohistochemically. After superovulation, eNOS levels increased progressively as the time went on. Moreover, the greatest amount of eNOS was observed in ovaries at the luteal phase. eNOS is considered as a constitutively expressed enzyme, indeed, ovarian eNOS exhibit a distinct, hormone-regulated expression pattern during follicular development, ovulation, and luteal phase in this mouse model.
Study on women in reproductive age:
In normally cycling women and stimulated patients, the highest plasma concentrations of NO metabolites corresponded to the midcycle, when the serum levels of estrogen were also greatest. This supports the existence of an E2 control of NO generation and release.
Conclusions
These data demonstrate that rat ovarian eNOS expression is regulated during folliculogenesis. eNOS was localized to the theca cell layer and stroma cells and the surface of oocytes in control ovaries. During follicular development, increased staining was observed in both theca cell layer and granulosa cells. At ovulation, no further changes in expression of eNOS were detected, but enhanced eNOS staining was observed within the CL. In Western blot analysis of eNOS in mouse ovaries, eNOS levels increased progressively after superovulation, and the greatest amount of eNOS was observed at the luteal phase. It is shown that the time-course changes of eNOS expression are induced by controlled ovarian hyperstimulation. However, the mechanism by which eNOS is induced in the ovary by gonadotropins is still unclear.
Both the highest plasma concentrations of NO metabolites and the greatest serum levels of estrogen correspond to the late follicular phase. These facts indicate that marked systemic vasodilator effects induced by estrogen may be partially mediated by nitric oxide. Improved ovarian and uterine blood flows are beneficial to ovulation and endometrial development. Furthermore, the reduction in NO metabolites observed in secretory phase suggests a possible opposing action of progesterone on NO generation or release. Further studies are needed to verify this inference.
誌謝 ----------------------------------------------------- p.01
中文摘要 ------------------------------------------------- p.02
目錄 ----------------------------------------------------- p.05
表目錄 --------------------------------------------------- p.06
圖目錄 --------------------------------------------------- p.07
圖表附錄 ------------------------------------------------- p.09
符號與縮寫 ----------------------------------------------- p.10
主文
I前言 ------------------------------------------- p.12
II材料 ------------------------------------------ p.25
III方法 ----------------------------------------- p.28
IV結果〈含圖表〉 -------------------------------- p.41
V討論 ------------------------------------------- p.63
VI參考文獻 -------------------------------------- p.75
VII英文摘要 ------------------------------------- p.80
作者簡歷 ------------------------------------------------- p.83
著作權聲明 ----------------------------------------------- p.84
1.Moncada S, Palmer RM, and Higgs EA, (1991) Nitric oxide: physiology, pathophysiology, and pharmacology. Pharmacol Rev 43:109-142.
2.Nathan C, (1992) Nitric oxide as a secretory product of mammalian cells. FASEB J 6:3051-3064.
3.Ragsdale RO, (1973) In Developments in Inorganic Nitrogen Chemistry, Vol. 2 (Colburn CB, ed.), Elsevier, Amsterdam, pp.1-26.
4.Mitchell HH, Shonie HA and Grindley HS, (1916) The origin of the nitrates in the urine. J. Biol. Chem., 24, 461-490.
5.Tannenbaum, SR, Fett, D Young, VR, (1928) Nitrite and nitrate are formed by endogenous synthesis in the human intestine. Science, 200, 1487-1489.
6.Furchgott RF and Zawadski JV, (1980) The obligatory role of the endothelial cells in the relaxation of arterial smooth muscle by acetylcholine. Nature, 288,373-376.
7.Palmer RMJ, Ferrige, AG and Moncada S, (1987) Nitric oxide release accounts for the biological activity of endotheliurn-derived relaxation factor. Nature, 327,524-526.
8.Ignarro LJ, Buga GM, Wood KS, (1987) Endothelium-derived relaxing factor produced and released from artery and vein is nitric oxide. Proc. Natl. Acad. Sci. USA, 84, 9265-9269.
9.Palmer RMJ, Ashton DS, and Moncada S, (1988) Vascular endothelial cells synihesize nitric oxide from L-arginine. Nature, 33, 664-668.
10.Griffith OW, and Stuehr DJ, (1995) Nitric oxide synthase properties and catalytic mechanism. Anna. Rev. Physiol., 57, 707-736.
11.Snyder SH, (1995) Nitric oxide: NO endothelial NO. Nature, 377, 196-197.
12.Nussler AK, and Billiar TR, (1993) Inflammation, immunoregulation and inducible nitric oxide synthase. J. Leukocyte Biol., 54, 171-178.
13.Morris SM, and Billiar TR, (1994) New insights into the regulation of inducible nitric oxide synthesis. Am. J. Physiol., 266, E829-E839.
14.Dinerman JL, Lowenstein LJ and Snyder SH, (1993) Molecular mechanisms of nitric oxide regulation: potential relevance to cardiovascular disease. Circ. Res., 73, 217-222.
15.Cho HJ, Martin E, Xie QW et al., (1995) Inducible nitric oxide synthase identification of amino acid residues essential for dimerization and binding of tetrahydrobiopterin. Proc. Nail. Acad. Sci. USA, 92, 11514-11518.
16.Wang J, Brown MA, Tam SH, Chan MC, Whitworth JA, (1997) Effects of diet on measurement of nitric oxide metabolites. Clin. Experi. Pharma. Physio., 24, 418-20.
17.Moncada S, (1992) The L-arginine: nitric oxide pathway. Acta Physiol Scand.; 145: 201-227.
18.Feelisch M, Stamler JS. (1996), Methods in Nitric Oxide Research. John Wiley & Sons Ltd, pp. 22.
19.White MM, Zamudio S, Stevens T, Tyler R, Lindenfeld J, Leslie K, Moore LG. (1995) Estrogen, progesterone, and vascular reactivity: potential cellular mechanisms. Endocr Rev; 16 (6):739-51.
20.Beckman JS, Beckman TW, Chen J et al., (1990) Apparent hydroxyl radical production of peroxynitrite: implications for endothelial injury from nitric oxide and superoxide. Proc. Natl. Acad. Sci. USA, 87, 1620-1624.
21.Beckman JS and Crow JP, (1993) Pathological implications of nitric oxide, superoxide and peroxynitrite formation. Biochem. Soc. Trans., 21, 330-334.
22.Rosselli M, Keller PJ, Dubey RK, (1998) Role of nitric oxide in the biology, physiology and pathophysiology of reproduction. Hum Reprod Update; 4(1): 3-24.
23.Mclachlan RI, Robertson DM, Healy DL, Burger HG and Kretser DM (1987) Circulating immunoreactive inhibin levels during the normal human menstrual cycle. J Clin Endocrinol Metab 65, 954-61.
24.Ellman C, Corbett JA, Misko TP, McDaniel M. Beciterman KP (1993) Nitric oxide mediates interleukin-lβ induced cellular cytotoxicity. A potential role for nitric oxide in the ovulatory process. J Clin Invest 92:3053-3056.
25.Van Voorhis BJ, Dunn MS, Snyder GD, and Weiner CP (1994) Nitric oxide: an autocrine regulator of human granulosa-luteal cell steroidogenesis. Endocrinology 135:1799-1806.
26.Ben-Shlomo I, Kokia E, Jackson MJ, Adashi EY, Payne DW, (1994) Interleukin-lβ stimulates nitrite production in the rat ovary; evidence for heterologous cell-cell interaction and for insulin-mediated regulation of the inducible isoform of nitric oxide synthase. Biol Reprod 51:310-318.
27.Shukovski L, Tsafriri A, (1994) The involvement of nitric oxide in the ovulatory process in the rat. Endocrinology 135:2287-2290.
28.Chun S-Y, Eisenhauer KM, Kubo M, Hsueh AJW, (1995) Interleukin-lβ suppresses apoptosis in rat ovarian follicles by increasing nitric oxide production. Endocrinology 136:3120-3127.
29.Powers RW, Chen L, Russell PT, Larsen WJ, (1995) Gonadotropin-stimulated regulation of blood-follicle barrier is mediated by nitric oxide. Am J Physiol 269:E290-E298.
30.Hogan B, Costantini F, Lacy E, (1986) Manipulating the Mouse Embryo: A Laboratory Manual. Cold Spring Harbor Laboratory 1986, New York, USA.
31.Purcell TL, Given R, Chwalisz k, & Garfield RE, (1999) Nitric oxide synthase distribution during implatation in the mouse. Mol Hum Repro 1999, 5(5): 467-75.
32.Tsai HD, Chen CM, Lo HY, Chang CC, (1995) Subcutaneous low does Leuprolide Acetate Depot Versus Leuprolide Acetate for Women Undergoing Ovarian Stimulation for in-Vitro Fertilization. Hum. Reprod., 1995; 10,2909~2912.
33.Yen SC, Jaffe RB, Barbieri RL, (1999) Reproductive Endocrinology: physiology, pathology, and clinical management. 4th Edition 1999, Saunders, USA.
34.Shoham Z, Mannaerts B, Insler V, (1993) Induction of follicular growth using recombinant follicle-stimulation hormone in two volunteer women with hypogonadotropic hypogonadism. Fertil Steril 59: 738-42.
35.Schoot DC, Coelingh-Bennink HJT, Mannaerts BM, (1992) Human recombinant follicle-stimulation hormone induces growth of preovulatoty follicles without concomitant increase in androgen and estrogen biosynthesis in a woman with isolated gonadotropin deficiency. J Clin Endocrinol Metab 74: 1471-73.
36.Van Voorhis BJ, Moore K, Strijbos PJL, Nelson S, Baylis SA, Grzybicki D, Weiner CP (1995) Expression and localization of inducible and endothelial nitric oxide synthase in the rat ovary. J Clin Invest 96:2719-2726.
37.North AJ, Star RA, Brannon TS, Ujiie K, Wells LB, Lowenstein CJ, Snyder SH, Shanl PW (1994) Nitric oxide synthase type I and type III gene expression are developmentally regulated in rat lung. Am J Physiol 266:L635-L641
38.Ghabour MS, Eis ALM, Brockman DE, Pollock S, Myatt L (1995) Immunohistochemical characterization of placental nitric oxide synthase expression in preeclampsia. Am J Obstet Gynecol 173:687-694.
39.Powers RW, Chambers C, Larsen W (1996) Diabetes-mediated decreases in ovarian superoxide dismutase activity are related to blood-follicle barrier and ovulation defects. Endocrinology 137:3101-3110.
40.Weiner CP, Uzasoam I, Baylis SA, Knowles RG, Charles IG, Moncada S (1994) Induction of calcium-dependent nitric oxide synthases by sex hormones. Proc Natl Acad Sci USA 91:5212-5216.
41.Zackrisson U, Mikuni M, Wallin A, Delbro D, Hedin L Brannstrom M, (1996) Cell-specific localization of nitric oxide synthases (NOS) in the rat ovary during follicular development, ovulation and luteal formation. Hum Reprod; 11(12): 2667-73.
42.Jablonka-Shariff A, Olson LM, (1997) Hormone regulation of nitric oxide synthases and their cell-specific expression during follicular development in the rat overy. Endocrinology: 138(1): 460-8.
43.Amet UA, McMillan A, Dinerman JL, Ballernann B, Lowenstein CJ (1996) Regulation of endothelial nitric-oxide synthase during hypoxia. J Biol Chem 271:15060-15073.
44.Stefano GB, Prevot V, Beauvillain JC, Cadet P, Fimiani C, Welters I, Fricchione GL, Breton C, Lassalle P, Salzet M, Bilfinger TV, (2000) Cell-surface estrogen receptors mediate calcium-dependent nitric oxide release in human endothelia. Circulation 2000 Apr 4; 101 (13):1594-7.
45.Stefano GB, Cadet P, Breton C, Goumon Y, Prevot V, Dessaint JP, Beauvillain JC, Roumier AS, Welters I, Salzet M,(2000) Estradiol-stimulated nitric oxide release in human granulocytes is dependent on intracellular calcium transients: evidence of a cell surface estrogen receptor. Blood 2000 Jun 15; 95 (12): 3951-3958
46.Nakamura Y, Kashida S, Nakata M, Takiguchi S, Yamagata Y, Takayama H, Sugino N, Kato H, (1999) Changes in nitric oxide synthase activity in the ovary of gonadotropin treated rats: the role of nitric oxide during ovulation. Endocr J 1999 Aug; 46(4):529-38.
47.Jablonka-Shariff A, Basuray R, Olson LM, (1999) Inhibitors of nitric oxide synthase influence oocyte maturation in rats. J Soc Gynecol Investig 1999 Mar-Apr; 6(2): 95-101.
48.Tsafriri A, Reich R, (1999) Molecular aspects of mammalian ovulation. Exp Clin Endocrinol Diabetes 107: 1-11.
49.Szego CM, Gitin ES, (1964) Ovarian histamine depletion during acute hyperaemic response to luteinizing hormone. Nature 201: 682-684.
50.Poweis RW, Chen L, Russell PT, Larsen WJ, (1995) Gonadotropin-stimulated regulation of blood-follicle barrier is mediated by nitric oxide. Am J Physiol 269: E290-E298.
51.Yamauchi J, Miyazaki T, Iwasaki S, Kishi I, Kuroshima M, Tei C, Yoshimura Y, (1997) Effexts of nitric oxide on ovulation and ovarian steroidogenesis and prostaglandin production in the rabbit. Endocrinology 138: 3630-3637.
52.Gaede SD, Sholley MM, Quattropani SL, (1985) Endothelial mitosis during the intial stages of corpus luteum neovascularization in the cycling adult rat. Am J Anat 172: 173-180.
53.Reynolds LP, Grazul-Bilska AT, Redmer DA, (2000) Angiogenesis in the corpus luteum. Endocrine 12(1):1-9.
54.Olson LM, Jones-Burton CM, Jablonka-Shariff A, (1996) Nitric oxide decreases estradiol synthesis in rat luteinized ovarian cells: possible role for nitric oxide in functional luteal regression. Endocrinology 137:3531-3539.
55.Snyder GD, Holmes RW, Bates Jn, Van Voorhis BJ, (1996) Nitric oxide inhibits aromatase activity: mechanisms of action. J Steroid Biochem mol Biol 58(1): 63-69.
56.Motta AB, Estevez A, de Gimeno MF, (1999) The invovement of nitric oxide in corpus luteum regession in the rat: feedback mechanism between prostaglandin F2α and nitric oxide. Mol Hum Reprod 5(11): 1011-1016.
57.Motta AB, Gimeno MA, (1997) Nitric oxide participate in the corpus luteum regression in ovaries isolated from pseudopregnant rats. Can J Physiol Pharmacol 75(12): 1335-1339.
58.Gobbetti A, Boiti C, Canali C, Zerani M, (1999) Nitric oxide synthase acutely regulates progesterone production by in vitro cultured rabbit corpora lutea. J Endocrinol 160(2): 275-283.
59.Boiti C, Zerani M, Zamponi D, Gobbetti A, (2000) Nitric oxide synthase activity and progesterone release by isolated corpora lutea of rabbits in the early and mid-luteal phases of pseudopregnancy are modulated differently by prostaglandin E2 and prostaglandin F2α via adenylate cyclase and phospholipase C.
60.Chun S-Y, Eisenhaoer KM, Knbo M, Hsueh AJW, (1995) interleukin-1βsuppresses apoptosis in rat ovarian follicles by increasing nitric oxide production. Endocrinology 136:3120-3127.
61.Beckman JS, Ye YZ, Anderson PG, Chen J Accavitti MA, Tarpey MM, White CR, (1994) Extensive nitration of protein tyrosines in human artherosclerosis detected by immunohistochemistry. Biol Chem Hoppe-Seyler 375: 81-88.
62.Yun H-Y, Dawson VL, Dawson TM, (1996) Neurobiology of nitric oxide. Crit Rev Neurobiol 10: 291-316.
63.Van Nassauw L, Tao L, Harrisson F, (1999) Localization of nitric oxide-related substances in the quail ovary during folliculogenesis. Histochem J 31(7): 443-454.
64.Feelisch M, Stamler JS, (1996) Methods in Nitric Oxide Research. John Wiley & Sons Ltd, pp. 55.
65.Rosselli M, Imthurm B, Macas E, Keller, Dubey RK, (1994) Circulating nitrite/nitrate levels increase with follicular development: inderct evidence for estradiol mediated NO release. Biochem Biophys Res Commun 202(3): 1543-1552.
66.Hayashi T, Yamada K, Esaki T, Kuzuya M, Satake S, Ishikawa T, Hidaka H, Iguchi A, (1995) Estrogen increases endothelial nitric oxide by a receptor-mediated system. Biochem Biophys Res Commun 214(3): 847-855.
67.Dickey RP, (1997) Doppler ultrasound investigation of uterine and ovarian blood flows in infertility and early pregnancy. Hum Reprod Update 3(5): 467-503.
68.Sher G, D.Fisch J, (2000) Vaginal sildenafil (Viagra): a preliminaty report of a novel method to improve uterine artery blood flow and endometrial development in patients undergoing IVF. Hum Reprod 15(4): 806-809.
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