小鼠睾丸引带细胞培养及已烯雌酚对培养睾丸引带细胞的影响
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摘要
目的:
许多研究表明,人类精子数和精液量在明显下降,男性生殖系统分化和发育异常则明显增加,同时环境雌激素(EEs)对男性生殖系统的影响也日益明显,且大多数与睾丸发育、下降不全相关,而睾丸引带与睾丸发育、下降关系密切。目前这方面还有许多问题尚未研究清楚,特别的是在EEs的作用机理方面。本实验通过建立小鼠睾丸引带细胞培养方案,利用经典的EEs己烯雌酚(DES)直接作用于培养的小鼠睾丸引带细胞,观察细胞的形态结构变化,并研究DES对培养小鼠睾丸引带细胞增殖和收缩活性等的影响。以探讨外源性雌激素(EEs)影响睾丸下降的可能机理。
材料和方法:
小鼠睾丸引带细胞原代培养
脱颈椎处死3d大雄性昆明小鼠,消毒后正中剪开下腹部,再次辨认雌雄,在3倍手术放大镜下于膀胱两侧游离出完整睾丸、附睾及睾丸引带,置于PBS中剔除出大部分睾丸引带组织。将引带组织放入含Ⅰ型胶原酶(1mg/ml)的DMEM培养液中,37℃持续震荡消化1小时后,加入3-4倍含无雌激素血清的培养液终止消化作用,静置后取上清液用25μm孔径过滤器过滤,1000r/min离心滤液5min,弃上清。细胞沉淀加入含无雌激素胎牛血清(10%v/v)DMEM培养基,吹打成单个细胞悬液,接种于25cm培养瓶中,置于5% CO2、37℃饱和湿度孵育箱中静置培养。根据不同的培养情况将帖壁的睾丸引带细胞分别传代接种到6孔板和96孔板中。台盼蓝染色计数法确定细胞存活率。
DES对小鼠睾丸引带细胞的影响
将1ml 4×104/ml的细胞悬液(原代细胞)接种于六孔板中,血清浓度为5%。细胞接种约24小时后,分实验组(DES 0.01μg/ml、0.10μg/ml、1.00μg/ml、10.00μg/ml组)、溶剂对照组(DMSO 1.00μl/ml)和正常组处理细胞。加药后不同时间(12h、24h和48h)获取标本,分别用CCK-8、Western Blot和细胞免疫化学等方法检测小鼠睾丸引带细胞中核增殖抗体(PCNA)、雌激素受体(ERα和ERβ)、F-actin和胞内钙离子浓度([Ca2+]i)的表达情况。
结果:
1.培养的小鼠睾丸引带细胞大部分为成纤维样细胞,有少数的上皮样细胞。细胞呈放射状、火焰状或漩涡状走行,胞体呈梭形或不规则三角形,胞质有突起。传代后细胞仍呈成纤维细胞型生长,同源性好,存活率为85%～90%。
2.实验组细胞随DES剂量与作用时间生长明显受抑制,细胞生长缓慢,突起回缩,相互间隙增大,细胞轮廓明显,胞质粗糙,内有颗粒堆积,DES剂量越高,细胞越失去成纤维细胞的形态。
3. CCK-8方法发现正常组细胞呈持续性增殖,于培养24h后增殖越趋明显。对照组、0.01μg/ml、0.10μg/ml和1.00μg/ml组增殖趋势一致,且数值接近,同一时间段与正常组比较无明显的差异(P>0.05),而同一组别各时间段与各自12h比较有明显的差异性(P<0.05)。DES10.00μg/ml组于12h开始持续呈半数抑制状态,与其他各组比较有明显的差异(P<0.05)。
4.细胞免疫荧光方法发现PCNA严格表达于细胞核中,Western Blot检测10.00μg/ml组的PCNA表达与其他各组比较有明显的差异(P<0.05),与CCK-8结果大致相同。
5.细胞免疫荧光方法发现睾丸引带细胞的F-actin在DES作用后发生明显的解聚,细胞骨架重塑,细胞周边肌动蛋白丝带模糊,应力纤维增加,在融合的细胞单层,粗而长的应力纤维相互交错,结构紊乱。且呈剂量和时间相关性。
6.小鼠睾丸引带细胞负载Flou-3荧光探针后在激光扫描电镜下检测[Ca2+]i,发现胞内荧光强度呈DES浓度依赖性增强,DES10.00μg/ml组甚至出现胞内钙的荧光强度呈持续性的高度增强。
7.细胞免疫化学发现小鼠睾丸引带细胞上存在ERα和ERβ,ERα表达于胞核,ERβ既表达于胞核又表达于胞浆。
结论:
1.成功建立了小鼠睾丸引带细胞培养方法。睾丸引带细胞呈成纤维细胞型生长,传代后细胞同源性好,存活率高。
2. DES可导致小鼠睾丸引带细胞形态异常、结构紊乱,且与剂量关系密切。
3. CCK-8和对PCNA的检测结果都显示DES对小鼠睾丸引带细胞有显著的细胞毒性作用,呈剂量-时间效应。
4. F-actin和[Ca2+]i的检测结果提示DES可能通过抑制小鼠睾丸引带细胞的收缩来影响睾丸的正常下降。除经典的胞内受体介导的信息传导途径外,EEs快速影响睾丸引带细胞内各信使的表达还可能通过非基因调控信号途径。
5.小鼠睾丸引带细胞存在ERα和ERβ,为EEs作用睾丸引带提供了理论基础。
Objective:Recently, there is a positive correlation between the environmental factors and high incidence of congenital disorders of the urinary system, especially the environment estrogens(EEs). However, aside from male sexual differentiation and the development of testicular tissue, the mechanism of testicular descent remains controversial. The gubernaculums plays an essential role in the complex mechanism of testicular descent, but its mechanism also remains unclear. However, although a large amount of international research has taken place on the EEs disruption issue over the last decade, the precise cellular and molecular mechanisms underlying the action of estrogen on the testis and gubernaculums testis are still largely obscure. The cultured mouse gubernacular cells were treated with the synthetic estrogen diethylstilbestrol (DES) in this experiment. And then the morphology and functions of the cells were observed or measured. Not only the mechanism of how time-and dose-related effects of EEs on the gubernacular cells were explored, but also, more important, the idea of research and the method of precise cell and molecular biology on testis descent or even the congenital genital abnormalities.
Methods: Gubernacula from 3-day-old mice were removed by operating magnifier and placed into phenol red-free Dulbecco’s modified Eagle’s medium (DMEM) supplemented with an enzyme solution containing typeⅠcollagenase ( 1mg/ml). The cultures were grown in 5% CO2 and 95% air in phenol red-free DMEM containing 5% charcoal dextran-treated FBS. After 3~5 days , primary cells were transferred into each wall of a 6-well culture plate. Cell viability was measured by trypan blue. Cell morphology was observed by HE stain. Subculturing cells were devided into six groups randomly, that is, normal group (not given any drug), control group (given DMSO,1μl/ml), and experimental groups 1~4: given 0.01μg/ml、0.10μg/ml、1.00μg/ml、10.00μg/ml DES respectively (DES dissolved in DMSO). After be treated 12h, 24h, 48h, the gubernacular cells were harvested to observe the morphology, proliferating and expression of PCNA, ER, F-actin, [Ca2+] I by Western Blot and cell immunity assay.
Results: 1~2h after incubation, the primary cells had appeared to adhere to the wall. On days 3~5, a confluent monolayer of fibroblasts had formed, mainly were fibroblasts, and a few epithelioid. Fibroblasts were closely arranged, radial, knitted or circinate. 1~2 days after subculturing, the fibroblasts adhere the wall completely. The staining of HE of fibroblasts displayed the culture cells were highly homogeneous morphology. The cell viability was about 85-90%. DES can induce transformation of gubernacular cells evidently and the proliferation of experimental groups cells were inhibited by different dosage of DES and different measuring time in some degree, especially the high dose. The ER (ERα&ERβ) exited in the mouse gubernacular cells, both expressed in the cell nucleus and ERβalso expressed in the cytoplasm. The expression of [Ca2+] I and PCNA were obvious dosage-effect and time-effect correlations(P<0.05). Normal and Control cells exhibited short processes and a diffuse filamentous staining with FITC-Phalloidin around the cell periphery and nucelus. Cells exposed to DES exhibited the stress fibres distributed throughout the cytoplasm gradually instead of the F-actin around the cells, especially the high dose.
Conclusion: The method of mouse gubernacular cell culture is established successfully. It can be applied to further investigation. DES is one of the classic EEs, the direct action in mouse gubernacular cells by DES can directly and accurately detect the dosage-effect and time-effect correlations of the expression of [Ca2+] I , PCNA and F-actin in gubernaculum testis. The ER proteins were abundant in the mouse gubernacular cells. It is proposed that EEs could lead to decline the contractive activity and inhibited the cell proliferation of gubernaculum, the effect androgen on gubernaculum testis was declined too. EEs may cause the anomogenesis and inhibition of descent of the mice gubernaculum testis or even testis.
许多研究表明,人类精子数和精液量在明显下降,男性生殖系统分化和发育异常则明显增加,同时环境雌激素(EEs)对男性生殖系统的影响也日益明显,且大多数与睾丸发育、下降不全相关,而睾丸引带与睾丸发育、下降关系密切。目前这方面还有许多问题尚未研究清楚,特别的是在EEs的作用机理方面。本实验通过建立小鼠睾丸引带细胞培养方案,利用经典的EEs己烯雌酚(DES)直接作用于培养的小鼠睾丸引带细胞,观察细胞的形态结构变化,并研究DES对培养小鼠睾丸引带细胞增殖和收缩活性等的影响。以探讨外源性雌激素(EEs)影响睾丸下降的可能机理。
材料和方法:
小鼠睾丸引带细胞原代培养
脱颈椎处死3d大雄性昆明小鼠,消毒后正中剪开下腹部,再次辨认雌雄,在3倍手术放大镜下于膀胱两侧游离出完整睾丸、附睾及睾丸引带,置于PBS中剔除出大部分睾丸引带组织。将引带组织放入含Ⅰ型胶原酶(1mg/ml)的DMEM培养液中,37℃持续震荡消化1小时后,加入3-4倍含无雌激素血清的培养液终止消化作用,静置后取上清液用25μm孔径过滤器过滤,1000r/min离心滤液5min,弃上清。细胞沉淀加入含无雌激素胎牛血清(10%v/v)DMEM培养基,吹打成单个细胞悬液,接种于25cm培养瓶中,置于5% CO2、37℃饱和湿度孵育箱中静置培养。根据不同的培养情况将帖壁的睾丸引带细胞分别传代接种到6孔板和96孔板中。台盼蓝染色计数法确定细胞存活率。
DES对小鼠睾丸引带细胞的影响
将1ml 4×104/ml的细胞悬液(原代细胞)接种于六孔板中,血清浓度为5%。细胞接种约24小时后,分实验组(DES 0.01μg/ml、0.10μg/ml、1.00μg/ml、10.00μg/ml组)、溶剂对照组(DMSO 1.00μl/ml)和正常组处理细胞。加药后不同时间(12h、24h和48h)获取标本,分别用CCK-8、Western Blot和细胞免疫化学等方法检测小鼠睾丸引带细胞中核增殖抗体(PCNA)、雌激素受体(ERα和ERβ)、F-actin和胞内钙离子浓度([Ca2+]i)的表达情况。
结果:
1.培养的小鼠睾丸引带细胞大部分为成纤维样细胞,有少数的上皮样细胞。细胞呈放射状、火焰状或漩涡状走行,胞体呈梭形或不规则三角形,胞质有突起。传代后细胞仍呈成纤维细胞型生长,同源性好,存活率为85%～90%。
2.实验组细胞随DES剂量与作用时间生长明显受抑制,细胞生长缓慢,突起回缩,相互间隙增大,细胞轮廓明显,胞质粗糙,内有颗粒堆积,DES剂量越高,细胞越失去成纤维细胞的形态。
3. CCK-8方法发现正常组细胞呈持续性增殖,于培养24h后增殖越趋明显。对照组、0.01μg/ml、0.10μg/ml和1.00μg/ml组增殖趋势一致,且数值接近,同一时间段与正常组比较无明显的差异(P>0.05),而同一组别各时间段与各自12h比较有明显的差异性(P<0.05)。DES10.00μg/ml组于12h开始持续呈半数抑制状态,与其他各组比较有明显的差异(P<0.05)。
4.细胞免疫荧光方法发现PCNA严格表达于细胞核中,Western Blot检测10.00μg/ml组的PCNA表达与其他各组比较有明显的差异(P<0.05),与CCK-8结果大致相同。
5.细胞免疫荧光方法发现睾丸引带细胞的F-actin在DES作用后发生明显的解聚,细胞骨架重塑,细胞周边肌动蛋白丝带模糊,应力纤维增加,在融合的细胞单层,粗而长的应力纤维相互交错,结构紊乱。且呈剂量和时间相关性。
6.小鼠睾丸引带细胞负载Flou-3荧光探针后在激光扫描电镜下检测[Ca2+]i,发现胞内荧光强度呈DES浓度依赖性增强,DES10.00μg/ml组甚至出现胞内钙的荧光强度呈持续性的高度增强。
7.细胞免疫化学发现小鼠睾丸引带细胞上存在ERα和ERβ,ERα表达于胞核,ERβ既表达于胞核又表达于胞浆。
结论:
1.成功建立了小鼠睾丸引带细胞培养方法。睾丸引带细胞呈成纤维细胞型生长,传代后细胞同源性好,存活率高。
2. DES可导致小鼠睾丸引带细胞形态异常、结构紊乱,且与剂量关系密切。
3. CCK-8和对PCNA的检测结果都显示DES对小鼠睾丸引带细胞有显著的细胞毒性作用,呈剂量-时间效应。
4. F-actin和[Ca2+]i的检测结果提示DES可能通过抑制小鼠睾丸引带细胞的收缩来影响睾丸的正常下降。除经典的胞内受体介导的信息传导途径外,EEs快速影响睾丸引带细胞内各信使的表达还可能通过非基因调控信号途径。
5.小鼠睾丸引带细胞存在ERα和ERβ,为EEs作用睾丸引带提供了理论基础。
Objective:Recently, there is a positive correlation between the environmental factors and high incidence of congenital disorders of the urinary system, especially the environment estrogens(EEs). However, aside from male sexual differentiation and the development of testicular tissue, the mechanism of testicular descent remains controversial. The gubernaculums plays an essential role in the complex mechanism of testicular descent, but its mechanism also remains unclear. However, although a large amount of international research has taken place on the EEs disruption issue over the last decade, the precise cellular and molecular mechanisms underlying the action of estrogen on the testis and gubernaculums testis are still largely obscure. The cultured mouse gubernacular cells were treated with the synthetic estrogen diethylstilbestrol (DES) in this experiment. And then the morphology and functions of the cells were observed or measured. Not only the mechanism of how time-and dose-related effects of EEs on the gubernacular cells were explored, but also, more important, the idea of research and the method of precise cell and molecular biology on testis descent or even the congenital genital abnormalities.
Methods: Gubernacula from 3-day-old mice were removed by operating magnifier and placed into phenol red-free Dulbecco’s modified Eagle’s medium (DMEM) supplemented with an enzyme solution containing typeⅠcollagenase ( 1mg/ml). The cultures were grown in 5% CO2 and 95% air in phenol red-free DMEM containing 5% charcoal dextran-treated FBS. After 3~5 days , primary cells were transferred into each wall of a 6-well culture plate. Cell viability was measured by trypan blue. Cell morphology was observed by HE stain. Subculturing cells were devided into six groups randomly, that is, normal group (not given any drug), control group (given DMSO,1μl/ml), and experimental groups 1~4: given 0.01μg/ml、0.10μg/ml、1.00μg/ml、10.00μg/ml DES respectively (DES dissolved in DMSO). After be treated 12h, 24h, 48h, the gubernacular cells were harvested to observe the morphology, proliferating and expression of PCNA, ER, F-actin, [Ca2+] I by Western Blot and cell immunity assay.
Results: 1~2h after incubation, the primary cells had appeared to adhere to the wall. On days 3~5, a confluent monolayer of fibroblasts had formed, mainly were fibroblasts, and a few epithelioid. Fibroblasts were closely arranged, radial, knitted or circinate. 1~2 days after subculturing, the fibroblasts adhere the wall completely. The staining of HE of fibroblasts displayed the culture cells were highly homogeneous morphology. The cell viability was about 85-90%. DES can induce transformation of gubernacular cells evidently and the proliferation of experimental groups cells were inhibited by different dosage of DES and different measuring time in some degree, especially the high dose. The ER (ERα&ERβ) exited in the mouse gubernacular cells, both expressed in the cell nucleus and ERβalso expressed in the cytoplasm. The expression of [Ca2+] I and PCNA were obvious dosage-effect and time-effect correlations(P<0.05). Normal and Control cells exhibited short processes and a diffuse filamentous staining with FITC-Phalloidin around the cell periphery and nucelus. Cells exposed to DES exhibited the stress fibres distributed throughout the cytoplasm gradually instead of the F-actin around the cells, especially the high dose.
Conclusion: The method of mouse gubernacular cell culture is established successfully. It can be applied to further investigation. DES is one of the classic EEs, the direct action in mouse gubernacular cells by DES can directly and accurately detect the dosage-effect and time-effect correlations of the expression of [Ca2+] I , PCNA and F-actin in gubernaculum testis. The ER proteins were abundant in the mouse gubernacular cells. It is proposed that EEs could lead to decline the contractive activity and inhibited the cell proliferation of gubernaculum, the effect androgen on gubernaculum testis was declined too. EEs may cause the anomogenesis and inhibition of descent of the mice gubernaculum testis or even testis.
引文
1. Yucel S, Cavalcanti AG, Desouza A, et al. The effect of oestrogen and testosterone on the urethral seam of the developing male mouse genital tubercle[J]. BJU 2003;92(9):1016-1021.
2. Rozati R, Reddy PP, Reddanna P, et al. Role of environmental estrogens in the deterioration of male factor fertility[J]. Fertil Steril 2002;78:1187-1194.
3. F.R.Boockfor, G.Fullbright,E.E. Büllesbach,et al. Relaxin-like factor (RLF) serum concentrations and gubernaculum RLF receptor display in relation to pre- and neonatal development of rats[J].Society for Reproduction and Fertility 2001;122:899-906.
4. Paris F, Jeandel C, Servant N, et al. Increased serum estrogenic bioactivity in three male newborns with ambiguous genitalia: a potential consequence of prenatal exposure to environmental endocrine disruptors[J]. Environ Res, 2006;100(1):39-43.
5. John M. Hutson1, Yasunari Sasaki, Jenny Huynh,et al. The gubernaculum in testicular descent and cryptorchidism[J]. The Turkish Journal of Pediatrics,2004; 46: 3-6.
6. Xue-Wu Jiang, Jian-Hong Li, Tian-Hua Huang, et al. Effect of prenatal exposure to diethylstilbestrol on gubernacular development in fetal male mice[J]. Asian J Androl 2004;6(4):325-329.
7. Gerald R. Cunha,1 John-Gunnar Forsberg,2 Robert Golden,et al. New Approaches for Estimating Risk from Exposure to Diethyistilbestrol[J].Environment Health Perspectives,1999;107:625-630.
8. Suh RS, Wang X, Ohl DA, et al. Influence of Prenatal Diethylstilbestrol (DES) and dietary phyto- estrogens on sperm function and reproductive organ development in male mice[J]. Fertility and Sterility 2003;80(Suppl 3):235.
9. Newbold RR, Jefferson WN, Padilla-Banks E, et al. Developmental exposure to diethylstilbestrol (DES) alters uterine response to estrogens in prepubescent mice: low versus high dose effects[J]. Reproductive Toxicology,2004;18(3):399-406.
10.Emmen JM, McLuskey A, Adham IM, et al. Involvement of insulin-like factor 3 (Insl3) in diethylstilbestrol-induced cryptorchidism[J]. Endocrinology. 2000;141(2):846-9.
11.McKinnll C, Sharpe RM, Mahood K, et al. Expression of insulin-like factor 3 protein in the rat testis during fetal and postnatal development and in relation to cryptorchidism induced by in utero exposure to di (n-Butyl) phthalate[J]. Endocrinology. 2005;146(10):4536-44.
12.Christophe Staub, Michel Rauch,Franc,et al. Expression of Estrogen Receptor ESR1 and Its 46-kDa Variant in the Gubernaculum Testis[J].BIOLOGY OF REPRODUCTION, 2005;73:703–712.
13.Sugita Y, Paxton GA, Hasthorpe S, et al. Does calcitonin gene-related peptide act as a chemoattractant for rat gubernacular cells[J]. J Pediatr Surg. 1997 Jan;32(1):15-7.
14.Carlsen E, Giwercman A, Keiding N, et al. Evidence for decreasing quality of semen during past 50 years[J]. Brmed J.1992.305(6854):609-613.
15.龚春雨,鲁刚,李虹,等.己烯雌酚建立尿道下裂大鼠模型的实验研究[J].中华小儿外科杂志,2004,25(2):180-181.
16.Inano H, Suzuki K, Onoda M, et al. Relationship between induction of mammary tumors and change of testicular functions in male rats following gamma-ray irradiation and/or diethylstilbestrol[J]. Carcinogenesis. 1996;17(2):355-360.
17.Klip H, Verloop J, van Gool JD, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study[J]. Lancet,2002,359(9312):1102-1107.
18.Troisi R, Hatch EE, Titus-Ernstoff L, et al. Cancer risk in women prenatally exposed to diethylstilbestrol[J]. Int J Cancer. 2007;121(2):356-360.
19.McLachlan J A, Wong A, Degen G H,et al. Morphologic and neoplastic transformation of Syrian hamster embryo fibroblasts by diethylstilbestrol and its analogs[J]. J Cancer-Res,1982; 42(8): 3040-3045.
20.Green S, Walter P, Greene G, et al. Cloning of the human oestrogen receptor cDNA[J]. J Steroid Biochem. 1986;24(1):77-83.
21.Kuiper GG, Enmark E, Pelto-Huikko M, et al. Cloning of a novel receptor expressed in rat prostate and ovary[J]. Proc Natl Acad Sci U S A. 1996;93(12):5925-5930.
22.Levin ER. Cellular Functions of the Plasma Membrane Estrogen Receptor[J].Trends Endocrinol Metab. 1999;10(9):374-377.
23.Adeoya-Osiguwa SA, Markoulaki S, Pocock V, et al. 17beta-Estradiol and environmental estrogens significantly affect mammalian sperm function[J].Hum Reprod. 2003;18(1):100-107.
24.Zsarnovszky A, Le HH, Wang HS,et al. Ontogeny of rapid estrogen-mediated extracellular signal-regulated kinase signaling in the rat cerebellar cortex: potent nongenomic agonist andendocrine disrupting activity of the xenoestrogen bisphenol A[J].Endocrinology. 2005;146(12):5388-5396.
25.Watson CS, Alyea RA, Jeng YJ,et al. Nongenomic actions of low concentration estrogens and xenoestrogens on multiple tissues[J]. Mol Cell Endocrinol. 2007;274(1-2):1-7.
26.Watson CS, Bulayeva NN, Wozniak AL,et al. Xenoestrogens are potent activators of nongenomic estrogenic responses[J]. Steroids. 2007;72(2):124-134.
27.Cowley SM, Hoare S, Mosselman S,et al. Estrogen receptors alpha and beta form heterodimers on DNA[J]. J Biol Chem. 1997;272(32):19858-19862.
28.Chen D, Washbrook E, Sarwar N,et al. Phosphorylation of human estrogen receptor alpha at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera[J]. Oncogene. 2002;21(32):4921-4931.
29.Saunders PT, Sharpe RM, Williams K,et al. Differential expression of oestrogen receptor alpha and beta proteins in the testes and male reproductive system of human and non-human primates[J]. Mol Hum Reprod. 2001;7(3):227-236.
30.Zhou Q, Nie R, Prins GS, et al. Localization of androgen and estrogen receptors in adult male mouse reproductive tract[J]. J Androl. 2002;23(6):870-881.
31.Bartlett JE, Washburn T, Eddy EM,et al. Early development of the gubernaculum and cremaster sac in estrogen receptor knockout mice[J]. Urol Res. 2001;29(3):163-167.
32.Staub C, Rauch M, Ferrière F,et al. Expression of estrogen receptor ESR1 and its 46-kDa variant in the gubernaculum testis[J]. Biol Reprod. 2005;73(4):703-712.
33.Cederroth CR, Schaad O, Descombes P, et al. Estrogen receptor alpha is a major contributor to estrogen-mediated fetal testis dysgenesis and cryptorchidism[J].Endocrinology. 2007;148(11):5507-5519.
34.Guillette LJ Jr, Gross TS, Masson GR, Matter JM, et al. Developmental abnormalities of the gonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida[J]. Environ Health Perspect. 1994;102(8):680-688.
35.Herbst AL, Ulfelder H, Poskanzer DC. Adenocarcinoma of the vagina:Association of maternal stilbestrol therapy with tumor appearance in young women[J]. N Engl J Med. 1971,284(15):878-881.
36.Guillette LJ Jr, Gross TS, Masson GR, Matter JM, et al. Developmental abnormalities of thegonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida[J]. Environ Health Perspect. 1994;102(8):680-688.
37.王金平,王滨有.尿道下裂危险因素的病例对照研究[J].中华流行病学杂志,2004,25(3):261-264.
38.Pierik FH, Burdorf A, Deddens JA, et al. Maternal and paternal risk factors for cryptorchidism and hypospadias: a case-control study in newborn boys[J]. Environ Health Perspect. 2004,112(15):1570-1576.
39.Shono T, Suita S, Kai H, et al. The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats[J]. J Pediatr Surg, 2004,39(2):213-216.
40.Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat[J]. Toxicol Sci,2000,58(2):350-365.
41.Virtanen HE, Rajpert-De Meyts E, Main KM,et al. Testicular dysgenesis syndrome and the development and occurrence of male reproductive disorders[J]. Toxicol Appl Pharmacol. 2005;207(2 Suppl):501-505.
42.Kaleva M, Virtanen HE, Haavisto AM,et al. Circannual rhythm in the incidence of cryptorchidism in Finland[J].Int J Androl. 2005;28(1):53-57.
43.Skakkebaek NE, J?rgensen N. Testicular dysgenesis and fertility[J]. Andrologia. 2005 ;37(6):217-218.
44.Lassurguère J, Livera G, Habert R, et al. Time- and dose-related effects of estradiol and diethylstilbestrol on the morphology and function of the fetal rat testis in culture[J]. Toxicol Sci. 2003;73(1):160-169.
45.Aravindakshan J, Paquet V, Gregory M,et al. Consequences of xenoestrogen exposure on male reproductive function in spottail shiners (Notropis hudsonius) [J]. Toxicol Sci. 2004;78(1):156-165.
46.Martinovi? D, Hogarth WT, Jones RE,et al. Environmental estrogens suppress hormones, behavior, and reproductive fitness in male fathead minnows[J]. Environ Toxicol Chem. 2007;26(2):271-8.
47.Doolan CM, Harvey BJ. A Galphas protein-coupled membrane receptor, distinct from the classical oestrogen receptor, transduces rapid effects of oestradiol on [Ca2+]i in female ratdistal colon[J]. Mol Cell Endocrinol. 2003;199(1-2):87-103.
48.Haynes MP, Li L, Sinha D,et al. Src kinase mediates phosphatidylinositol 3-kinase/Akt-dependent rapid endothelial nitric-oxide synthase activation by estrogen[J]. J Biol Chem. 2003;278(4):2118-2123.
49.Cunha GR, Forsberg JG, Golden R, et al. New approaches for estimating risk from exposure to diethylstilbestrol[J]. Environ Health Perspect. 1999 Aug;107 Suppl 4:625-30.
50.Bulayeva NN, Watson CS. Xenoestrogen-induced ERK-1 and ERK-2 activation via multiple membrane-initiated signaling pathways[J]. Environ Health Perspect. 2004;112(15):1481-1487.
51.Takeuchi A, Mishina Y, Miyaishi O,et al. Heterozygosity with respect to Zfp148 causes complete loss of fetal germ cells during mouse embryogenesis[J]. Nat Genet. 2003;33(2):172-176.
52.Maga G, Hubscher U. Proliferating cell nuclear antigen(PCNA), A dancer with many partners[J]. J Cell Sci, 2003;116(pt 15):3051-3060.
53.El Zoghbi CS, Favorito LA, Costa WS,et al. Structural analysis of gubernaculum testis in cryptorchid patients submitted to treatment with human chorionic gonadotrophin[J]. Int Braz J Urol. 2007;33(2):223-229.
54.Tanyel FC. The descent of testis and reason for failed descent[J]. Turk J Pediatr. 2004;46(Suppl):7-17.
55.Tanyel FC, Talim B, Atilla P,et al. Myogenesis within the human gubernaculum: histological and immunohistochemical evaluation[J]. Eur J Pediatr Surg. 2005;15(3):175-179.
56.蒋学武,邓汪东,胡显良等.外源性雌激素对小鼠睾丸引带收缩和增殖活性的影响[J].中华小儿外科杂志,2005;26(9):488-491.
57.Costa WS, Sampaio FJ, Favorito LA,et al. Testicular migration: remodeling of connective tissue and muscle cells in human gubernaculum testis[J]. J Urol. 2002;167(5):2171-2176.
58.Tomiyama H, Hutson JM. Contractility of rat gubernacula affected by calcitonin gene-related peptide and beta-agonist[J]. J Pediatr Surg. 2005;40(4):683-687.
59.Sasaki Y, Hutson JM, Clarke MC. The role of sympathetic innervation in the developing rat gubernaculum[J]. J Pediatr Surg. 2007;42(2):350-354.
1) Carlsen E, Giwercman A, Keiding N, et al. Evidence for decreasing quality of semen during past 50 years[J]. Brmed J.1992.305(6854):609-613.
2) Gerald R. Cunha,1 John-Gunnar Forsberg,2 Robert Golden,et al. New Approaches for Estimating Risk from Exposure to Diethyistilbestrol[J].Environment Health Perspectives,1999;107:625-630.
3) Herbst AL, Ulfelder H, Poskanzer DC. Adenocarcinoma of the vagina:Association of maternal stilbestrol therapy with tumor appearance in young women[J]. N Engl J Med. 1971,284(15):878-881.
4) Klip H, Verloop J, van Gool JD, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study[J]. Lancet,2002,359(9312):1102-1107.
5) Low-Dose Peer Review Panel(EPA,NIEHS and NIH-NTP).National Toxicology Proguam’s Report of the Endocine Disruptors Low-Dose Peer Review.2001,8,p53-54. http://ntp.niehs.nih.gov.
6) Suh RS, Wang X, Ohl DA, et al. Influence of Prenatal Diethylstilbestrol (DES) and dietary phyto- estrogens on sperm function and reproductive organ development in male mice[J]. Fertility and Sterility 2003;80(Suppl 3):235.
7) Newbold RR, Jefferson WN, Padilla-Banks E, et al. Developmental exposure to diethylstilbestrol (DES) alters uterine response to estrogens in prepubescent mice: low versus high dose effects[J]. Reproductive Toxicology,2004;18(3):399-406.
8) Emmen JM, McLuskey A, Adham IM, et al. Involvement of insulin-like factor 3 (Insl3) in diethylstilbestrol-induced cryptorchidism[J]. Endocrinology. 2000;141(2):846-9.
9)龚春雨,鲁刚,李虹,等.己烯雌酚建立尿道下裂大鼠模型的实验研究[J].中华小儿外科杂志,2004,25(2):180-181.
10) Inano H, Suzuki K, Onoda M, et al. Relationship between induction of mammary tumors and change of testicular functions in male rats following gamma-ray irradiation and/or diethylstilbestrol[J]. Carcinogenesis. 1996;17(2):355-360.
11) Troisi R, Hatch EE, Titus-Ernstoff L, et al. Cancer risk in women prenatally exposed to diethylstilbestrol[J]. Int J Cancer. 2007;121(2):356-360.
12) Green S, Walter P, Greene G, et al. Cloning of the human oestrogen receptor cDNA[J]. J Steroid Biochem. 1986;24(1):77-83.
13) Kuiper GG, Enmark E, Pelto-Huikko M, et al. Cloning of a novel receptor expressed in rat prostate and ovary[J]. Proc Natl Acad Sci U S A. 1996;93(12):5925-5930.
14) Levin ER. Cellular Functions of the Plasma Membrane Estrogen Receptor[J]. Trends Endocrinol Metab. 1999;10(9):374-377.
15) Adeoya-Osiguwa SA, Markoulaki S, Pocock V, et al. 17beta-Estradiol and environmental estrogens significantly affect mammalian sperm function[J]. Hum Reprod. 2003;18(1):100-107.
16) Zsarnovszky A, Le HH, Wang HS,et al. Ontogeny of rapid estrogen-mediated extracellular signal-regulated kinase signaling in the rat cerebellar cortex: potent nongenomic agonist and endocrine disrupting activity of the xenoestrogen bisphenol A[J]. Endocrinology. 2005;146(12):5388-5396.
17) Watson CS, Alyea RA, Jeng YJ,et al. Nongenomic actions of low concentration estrogens and xenoestrogens on multiple tissues[J]. Mol Cell Endocrinol. 2007;274(1-2):1-7.
18) Watson CS, Bulayeva NN, Wozniak AL,et al. Xenoestrogens are potent activators of nongenomic estrogenic responses[J]. Steroids. 2007;72(2):124-134.
19) Cowley SM, Hoare S, Mosselman S,et al. Estrogen receptors alpha and beta form heterodimers on DNA[J]. J Biol Chem. 1997;272(32):19858-19862.
20) Chen D, Washbrook E, Sarwar N,et al. Phosphorylation of human estrogen receptor alpha at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera[J]. Oncogene. 2002;21(32):4921-4931.
21) Saunders PT, Sharpe RM, Williams K,et al. Differential expression of oestrogen receptor alpha and beta proteins in the testes and male reproductive system of human and non-human primates[J]. Mol Hum Reprod. 2001;7(3):227-236.
22) Zhou Q, Nie R, Prins GS, et al. Localization of androgen and estrogen receptors in adult male mouse reproductive tract[J]. J Androl. 2002;23(6):870-881.
23) Bartlett JE, Washburn T, Eddy EM,et al. Early development of the gubernaculum and cremaster sac in estrogen receptor knockout mice[J]. Urol Res. 2001;29(3):163-167.
24) Staub C, Rauch M, Ferrière F,et al. Expression of estrogen receptor ESR1 and its 46-kDa variant in the gubernaculum testis[J]. Biol Reprod. 2005;73(4):703-712.
25) Cederroth CR, Schaad O, Descombes P, et al. Estrogen receptor alpha is a major contributorto estrogen-mediated fetal testis dysgenesis and cryptorchidism[J]. Endocrinology. 2007;148(11):5507-5519.
26) Guillette LJ Jr, Gross TS, Masson GR, Matter JM, et al. Developmental abnormalities of the gonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida[J]. Environ Health Perspect. 1994;102(8):680-688.
27) Paris F, Jeandel C, Servant N, et al. Increased serum estrogenic bioactivity in three male newborns with ambiguous genitalia: a potential consequence of prenatal exposure to environmental endocrine disruptors[J]. Environ Res, 2006;100(1):39-43.
28)王金平,王滨有.尿道下裂危险因素的病例对照研究[J].中华流行病学杂志,2004,25(3):261-264.
29) Pierik FH, Burdorf A, Deddens JA, et al. Maternal and paternal risk factors for cryptorchidism and hypospadias: a case-control study in newborn boys[J]. Environ Health Perspect. 2004,112(15):1570-1576.
30) Shono T, Suita S, Kai H, et al. The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats[J]. J Pediatr Surg, 2004,39(2):213-216.
31) Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat[J]. Toxicol Sci,2000,58(2):350-365.
32) Virtanen HE, Rajpert-De Meyts E, Main KM,et al. Testicular dysgenesis syndrome and the development and occurrence of male reproductive disorders[J]. Toxicol Appl Pharmacol. 2005;207(2 Suppl):501-505.
33) Kaleva M, Virtanen HE, Haavisto AM,et al. Circannual rhythm in the incidence of cryptorchidism in Finland[J]. Int J Androl. 2005;28(1):53-57.
34) Skakkebaek NE, J?rgensen N. Testicular dysgenesis and fertility[J]. Andrologia. 2005 ;37(6):217-218.
35) Lassurguère J, Livera G, Habert R, et al. Time- and dose-related effects of estradiol and diethylstilbestrol on the morphology and function of the fetal rat testis in culture[J]. Toxicol Sci. 2003;73(1):160-169.
36) Aravindakshan J, Paquet V, Gregory M,et al. Consequences of xenoestrogen exposure onmale reproductive function in spottail shiners (Notropis hudsonius) [J]. Toxicol Sci. 2004;78(1):156-165.
37) Martinovi? D, Hogarth WT, Jones RE,et al. Environmental estrogens suppress hormones, behavior, and reproductive fitness in male fathead minnows[J]. Environ Toxicol Chem. 2007;26(2):271-8.
38) McKinnll C, Sharpe RM, Mahood K, et al. Expression of insulin-like factor 3 protein in the rat testis during fetal and postnatal development and in relation to cryptorchidism induced by in utero exposure to di (n-Butyl) phthalate[J]. Endocrinology. 2005;146(10):4536-44.
39) Christophe Staub, Michel Rauch,Franc,et al. Expression of Estrogen Receptor ESR1 and Its 46-kDa Variant in the Gubernaculum Testis[J]. BIOLOGY OF REPRODUCTION, 2005;73:703–712.
40) Doolan CM, Harvey BJ. A Galphas protein-coupled membrane receptor, distinct from the classical oestrogen receptor, transduces rapid effects of oestradiol on [Ca2+]i in female rat distal colon[J]. Mol Cell Endocrinol. 2003;199(1-2):87-103.
41) Haynes MP, Li L, Sinha D,et al. Src kinase mediates phosphatidylinositol 3-kinase/Akt-dependent rapid endothelial nitric-oxide synthase activation by estrogen[J]. J Biol Chem. 2003;278(4):2118-2123.
42) Fraser LR, Beyret E, Milligan SR,et al. Effects of estrogenic xenobiotics on human and mouse spermatozoa[J]. Hum Reprod. 2006;21(5):1184-1193.
43) Brucker-Davis F, Thayer K, Colborn T. Significant effects of mild endogenous hormonal changes in humans: considerations for low-dose testing[J]. Environ Health Perspect. 2001;109(Suppl 1):21-26.
44) Bulayeva NN, Watson CS. Xenoestrogen-induced ERK-1 and ERK-2 activation via multiple membrane-initiated signaling pathways[J]. Environ Health Perspect. 2004;112(15):1481-1487.
[1]王光义,陈绍基,蒋学武,等.尿道下裂伴阴茎发育不良的内分泌改变及治疗[J].中华小儿外科杂志,1995;16(1):23-25.
[2]吴艳乔,王艳萍,朱军,等.国人尿道下裂的流行病学特征[J].中华泌尿外科杂志,1997,18(12):753-757.
[3]吴艳乔,代礼,王艳萍,等.中国儿童尿道下裂发生率的变化趋势[J].四川大学学报:医学版,2005,36(2):274-276.
[4]张伟,张波,邓兵.尿道下裂患儿性染色体异常的细胞及分子遗传学研究[J].中华实用医学,2004,6(16):4-6.
[5] Chu JH, Zhang RM, Zhao ZM, et al. Male Fertility Is Compatible with an Arg840Cys Substitution in the AR in a Large Chinese Family Affected with Divergent Phenotypes of AR Insensitivity Syndrome[J]. J Clin Endocrinol Metab,2002,87(1):347–351.
[6] Abbate B, Danti DA, Mattei R, et al. Genetic aspects of hypospadias: considerations apropos of 362 cases[J]. Pediatr Med Chir,1994,16(1):77-79.
[7] Kim KS, Liu W, Cunha GR, et al. Expression of the androgen receptor and 5alpha-reductase type 2 in the developing human fetal penis and urethra[J]. Cell Tissue Res,2002,307(2):145-153.
[8]贾悦,王晓东,崔毓桂,等. 5α-还原酶在男性生殖中的作用[J].生殖医学杂志,2003,12(2):115-118.
[9] Vilchis F, Mendez JP, Canto P,et al. Identification of missense mutations in the SRD5A2 gene from patients with steroid 5alpha-reductase 2 deficiency[J]. Clin Endocrinol (Oxf),2000 ,52(3):383-387.
[10] Silver R I, RussellDW. 5 alpha-reductase type 2mutations are present in some boys with isolated hypospadias[J]. J Urol ,1999, 162 (3 Pt2 ): 1142-1145.
[11]周李,梅骅,刘唐彬,等.先天性尿道下裂与SRD5A2及SRY基因突变关系研究[J].中华医学遗传学杂志,1999,16(5);311-314.
[12] Wang YP, Li Q, Xu JJ, et al. Mutation analysis of five candidate genes in Chinese patients with hypospadias[J]. Eur J Hum Genet ,2004,12(9):706-712.
[13]徐家杰,李森恺,李强,等.尿道下裂患者SRD5A2基因突变的研究[J].中华整形外科杂志,2006,22(2): 139-141.
[14] Aaronson IA, Cakmak MA, Key LL. Defects of the testosterone biosynthetic pathway in boys with hypospadias[J]. J Urol ,1997,157(5):1884-1888.
[15]唐建华,袁继炎.尿道下裂与Ⅱ型3β-羟类固醇脱氢酶基因突变的研究[J].临床小儿外科杂志,2002,1(4):282-285.
[16]李春,谢家伦,卢汉平.先天性尿道下裂外生殖器皮肤细胞内雄激素受体的含量测定[J].衡阳医学院学报, 1998,26(2):179-181.
[17]李强,李森恺,徐家杰,等.尿道下裂患者雄激素受体基因突变的研究[J].中华整形外科杂志,2004,20(6):421-424.
[18]徐哲,郑克立,乔慧,等.雄激素受体基因(CAG)n重复多态与先天性尿道下裂的关系[J].临床小儿外科杂志,2002,1(6):435-437.
[19] Aschim EL, Nordenskjold A, Giwercman A, et al. Linkage between cryptorchidism, hypospadias, and GGN repeat length in the androgen receptor gene[J]. J Clin Endocrinol Metab ,2004,89(10):5105-5109.
[20]蒋学武,陈中献,陈绍基,等.先天性尿道下裂患儿阴茎组织中多种性激素受体的比较研究[J].中国男科学杂志,2000,14 (1) :16-18.
[21] Castineyra G, Copelli S, Levalle O. 46, XX male: clinical, hormonal/genetic findings[J]. Arch Androl ,2002,48 (4) : 251-257.
[22] Quigley DI, McDonald MT, Krishnamuthy V,et al. Triploid mosaicism in a 45,X/69,XXY infant[J]. Am J Med Genet A. 2005, 138 ( 2 ) : 171-4.
[23]刘国庆,赵蕊,叶志纯,等.先天性尿道下裂与SRY基因关系的探讨[J].中华泌尿外科志,2001,22 (4) : 241-242.
[24]李强,李森恺,徐家杰.尿道下裂病人中SRY基因缺失的快速检测[J].中华整形外科杂志,2002 (S1) : 40-41.
[25] Moreno-Garcia M, Miranda EB. Chromosomal anomalies in cryptorchidism and hypospadias[J]. J Urol, 2002 ,168 ( 5 ) : 2170-2172.
[26]游慧灵,余敏. 14例尿道下裂患儿的细胞遗传学分析[J].中国优生与遗传志,2001,9(3):54,127.
[27] Paris F, Jeandel C, Servant N, et al. Increased serum estrogenic bioactivity in three male newborns with ambiguous genitalia: a potential consequence of prenatal exposure to environmental endocrine disruptors[J]. Environ Res, 2006,100 ( 1 ) : 39-43.
[28]罗艳,栾荣生,朱军,等.尿道下裂与环境污染的关系—Poisson回归模型与GM (1 , N)模型结合分析.现代预防医学[J], 2005 , 32 ( 1 ) : 1-2 , 15.
[29] Kun Suk Kim, Carlos R. Torres Jr., et al. Induction of hypospadias in a murine model by maternal exposure to synthetic estrogens[J]. J Environmental Research ,2004, 94 : 267–275.
[30] Willingham E, Agras K, Vilela M, et al. Loratadine Exerts Estrogen-Like Effects and Disrupts Penile Development in the Mouse[J]. J Urol, 2006,175 (2) :723-726.
[31]龚春雨,鲁刚,李虹,等.己烯雌酚建立尿道下裂大鼠模型的实验研究[J].中华小儿外科杂志,2004,25 (2) :180-181.
[32] Klip H, Verloop J, van Gool JD, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study[J]. Lancet , 2002,359 (9312) : 1102-1107.
[33] Kallen B. Use of antihistamine drugs in early pregnancy and delivery outcome[J]. J Matern Fetal Neonatal Med , 2002 ,11 (3) : 146-152.
[34] Brouwers MM, Feitz WF, Roelofs LA, et al. Hypospadias: a transgenerational effect of diethylstilbestrol[J] Hum Reprod, 2006,21 ( 3 ) : 666-669.
[35] Palmer JR, Wise LA, Robboy SJ, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero[J]. Epidemiology , 2005 ,16 ( 4 ) : 583-586.
[36] Centers for Disease Control and Prevention (CDC). Evaluation of an association between loratadine and hypospadias--United States, 1997-2001. MMWR Morb Mortal Wkly Rep, 2004,53(10):219-21.
[37]贺厚光,张炜,朱佳庚.小鼠尿道下裂动物模型的建立[J].中华男科学,2004,10 ( 3 ) : 172-174.
[38]韩雪冰,杨晓峰,任连生,等.先天性尿道下裂动物模型的建立[J].中华实验外科学杂志,2005, 22 ( 5 ) : 590.
[39]李岩,张浩,王文冬,等.小剂量氟他胺对大鼠性分化影响的实验研究[J].四川大学学报(医学版) , 2006 ,37 ( 2 ) : 258-261.
[40]张炜,袁琳,吴婷,等.邻苯二甲酸二丁酯诱导尿道下裂大鼠模型的建立及其作用机制[J].中华实验外科杂志, 2005 , 22 ( 2 ) : 246-248.
[41] Shono T, Suita S, Kai H, et al. The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats[J]. J Pediatr Surg, 2004, 39 ( 2) : 213-216.
[42] Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat[J]. Toxicol Sci , 2000 , 58 ( 2 ) : 350-365.
[43]王金平,王滨有.尿道下裂危险因素的病例对照研究[J].中华流行病学杂志, 2004 , 25 ( 3 ) : 261-264.
[44] Pierik FH, Burdorf A, Deddens JA, et al. Maternal and paternal risk factors for cryptorchidism and hypospadias: a case-control study in newborn boys[J]. Environ Health Perspect. 2004 , 112 ( 15 ) : 1570-1576.
[45] North K, Golding J. A maternal vegetarian diet in pregnancy is associated with hypospadias. The ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood[J]. BJU Int. 2000 , 85 (1) :107-113.
[46] LIU BC, WANG Z, LIN GT, et al. Activating Transcription Factor 3 Is Up-Regulated in Patients with Hypospadias[J]. Pediatric Research , 2005 ,58( 6) : 1280-1283 .
[47] Carmichael SL, Shaw GM, Laurent C, et al . Maternal progestin intake and risk of hypospadias[J]. Arch Pediatr Adolesc Med , 2005 , 159 ( 10 ) : 957-62.
[48] Silver RI, Rodriguez R, Chang TS, et a l. In vitro Fertilization is assosiated with an increased risk of hypospadias[J]. J Uro l, 1999 , 161 : 1954-1957.
[49]王军,袁继炎,周学锋.尿道下裂患儿包皮组织中表皮生长因子及其受体的表达及意义[J].中华小儿外科杂志, 2001 , 22 ( 1 ) : 30-31.
2. Rozati R, Reddy PP, Reddanna P, et al. Role of environmental estrogens in the deterioration of male factor fertility[J]. Fertil Steril 2002;78:1187-1194.
3. F.R.Boockfor, G.Fullbright,E.E. Büllesbach,et al. Relaxin-like factor (RLF) serum concentrations and gubernaculum RLF receptor display in relation to pre- and neonatal development of rats[J].Society for Reproduction and Fertility 2001;122:899-906.
4. Paris F, Jeandel C, Servant N, et al. Increased serum estrogenic bioactivity in three male newborns with ambiguous genitalia: a potential consequence of prenatal exposure to environmental endocrine disruptors[J]. Environ Res, 2006;100(1):39-43.
5. John M. Hutson1, Yasunari Sasaki, Jenny Huynh,et al. The gubernaculum in testicular descent and cryptorchidism[J]. The Turkish Journal of Pediatrics,2004; 46: 3-6.
6. Xue-Wu Jiang, Jian-Hong Li, Tian-Hua Huang, et al. Effect of prenatal exposure to diethylstilbestrol on gubernacular development in fetal male mice[J]. Asian J Androl 2004;6(4):325-329.
7. Gerald R. Cunha,1 John-Gunnar Forsberg,2 Robert Golden,et al. New Approaches for Estimating Risk from Exposure to Diethyistilbestrol[J].Environment Health Perspectives,1999;107:625-630.
8. Suh RS, Wang X, Ohl DA, et al. Influence of Prenatal Diethylstilbestrol (DES) and dietary phyto- estrogens on sperm function and reproductive organ development in male mice[J]. Fertility and Sterility 2003;80(Suppl 3):235.
9. Newbold RR, Jefferson WN, Padilla-Banks E, et al. Developmental exposure to diethylstilbestrol (DES) alters uterine response to estrogens in prepubescent mice: low versus high dose effects[J]. Reproductive Toxicology,2004;18(3):399-406.
10.Emmen JM, McLuskey A, Adham IM, et al. Involvement of insulin-like factor 3 (Insl3) in diethylstilbestrol-induced cryptorchidism[J]. Endocrinology. 2000;141(2):846-9.
11.McKinnll C, Sharpe RM, Mahood K, et al. Expression of insulin-like factor 3 protein in the rat testis during fetal and postnatal development and in relation to cryptorchidism induced by in utero exposure to di (n-Butyl) phthalate[J]. Endocrinology. 2005;146(10):4536-44.
12.Christophe Staub, Michel Rauch,Franc,et al. Expression of Estrogen Receptor ESR1 and Its 46-kDa Variant in the Gubernaculum Testis[J].BIOLOGY OF REPRODUCTION, 2005;73:703–712.
13.Sugita Y, Paxton GA, Hasthorpe S, et al. Does calcitonin gene-related peptide act as a chemoattractant for rat gubernacular cells[J]. J Pediatr Surg. 1997 Jan;32(1):15-7.
14.Carlsen E, Giwercman A, Keiding N, et al. Evidence for decreasing quality of semen during past 50 years[J]. Brmed J.1992.305(6854):609-613.
15.龚春雨,鲁刚,李虹,等.己烯雌酚建立尿道下裂大鼠模型的实验研究[J].中华小儿外科杂志,2004,25(2):180-181.
16.Inano H, Suzuki K, Onoda M, et al. Relationship between induction of mammary tumors and change of testicular functions in male rats following gamma-ray irradiation and/or diethylstilbestrol[J]. Carcinogenesis. 1996;17(2):355-360.
17.Klip H, Verloop J, van Gool JD, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study[J]. Lancet,2002,359(9312):1102-1107.
18.Troisi R, Hatch EE, Titus-Ernstoff L, et al. Cancer risk in women prenatally exposed to diethylstilbestrol[J]. Int J Cancer. 2007;121(2):356-360.
19.McLachlan J A, Wong A, Degen G H,et al. Morphologic and neoplastic transformation of Syrian hamster embryo fibroblasts by diethylstilbestrol and its analogs[J]. J Cancer-Res,1982; 42(8): 3040-3045.
20.Green S, Walter P, Greene G, et al. Cloning of the human oestrogen receptor cDNA[J]. J Steroid Biochem. 1986;24(1):77-83.
21.Kuiper GG, Enmark E, Pelto-Huikko M, et al. Cloning of a novel receptor expressed in rat prostate and ovary[J]. Proc Natl Acad Sci U S A. 1996;93(12):5925-5930.
22.Levin ER. Cellular Functions of the Plasma Membrane Estrogen Receptor[J].Trends Endocrinol Metab. 1999;10(9):374-377.
23.Adeoya-Osiguwa SA, Markoulaki S, Pocock V, et al. 17beta-Estradiol and environmental estrogens significantly affect mammalian sperm function[J].Hum Reprod. 2003;18(1):100-107.
24.Zsarnovszky A, Le HH, Wang HS,et al. Ontogeny of rapid estrogen-mediated extracellular signal-regulated kinase signaling in the rat cerebellar cortex: potent nongenomic agonist andendocrine disrupting activity of the xenoestrogen bisphenol A[J].Endocrinology. 2005;146(12):5388-5396.
25.Watson CS, Alyea RA, Jeng YJ,et al. Nongenomic actions of low concentration estrogens and xenoestrogens on multiple tissues[J]. Mol Cell Endocrinol. 2007;274(1-2):1-7.
26.Watson CS, Bulayeva NN, Wozniak AL,et al. Xenoestrogens are potent activators of nongenomic estrogenic responses[J]. Steroids. 2007;72(2):124-134.
27.Cowley SM, Hoare S, Mosselman S,et al. Estrogen receptors alpha and beta form heterodimers on DNA[J]. J Biol Chem. 1997;272(32):19858-19862.
28.Chen D, Washbrook E, Sarwar N,et al. Phosphorylation of human estrogen receptor alpha at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera[J]. Oncogene. 2002;21(32):4921-4931.
29.Saunders PT, Sharpe RM, Williams K,et al. Differential expression of oestrogen receptor alpha and beta proteins in the testes and male reproductive system of human and non-human primates[J]. Mol Hum Reprod. 2001;7(3):227-236.
30.Zhou Q, Nie R, Prins GS, et al. Localization of androgen and estrogen receptors in adult male mouse reproductive tract[J]. J Androl. 2002;23(6):870-881.
31.Bartlett JE, Washburn T, Eddy EM,et al. Early development of the gubernaculum and cremaster sac in estrogen receptor knockout mice[J]. Urol Res. 2001;29(3):163-167.
32.Staub C, Rauch M, Ferrière F,et al. Expression of estrogen receptor ESR1 and its 46-kDa variant in the gubernaculum testis[J]. Biol Reprod. 2005;73(4):703-712.
33.Cederroth CR, Schaad O, Descombes P, et al. Estrogen receptor alpha is a major contributor to estrogen-mediated fetal testis dysgenesis and cryptorchidism[J].Endocrinology. 2007;148(11):5507-5519.
34.Guillette LJ Jr, Gross TS, Masson GR, Matter JM, et al. Developmental abnormalities of the gonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida[J]. Environ Health Perspect. 1994;102(8):680-688.
35.Herbst AL, Ulfelder H, Poskanzer DC. Adenocarcinoma of the vagina:Association of maternal stilbestrol therapy with tumor appearance in young women[J]. N Engl J Med. 1971,284(15):878-881.
36.Guillette LJ Jr, Gross TS, Masson GR, Matter JM, et al. Developmental abnormalities of thegonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida[J]. Environ Health Perspect. 1994;102(8):680-688.
37.王金平,王滨有.尿道下裂危险因素的病例对照研究[J].中华流行病学杂志,2004,25(3):261-264.
38.Pierik FH, Burdorf A, Deddens JA, et al. Maternal and paternal risk factors for cryptorchidism and hypospadias: a case-control study in newborn boys[J]. Environ Health Perspect. 2004,112(15):1570-1576.
39.Shono T, Suita S, Kai H, et al. The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats[J]. J Pediatr Surg, 2004,39(2):213-216.
40.Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat[J]. Toxicol Sci,2000,58(2):350-365.
41.Virtanen HE, Rajpert-De Meyts E, Main KM,et al. Testicular dysgenesis syndrome and the development and occurrence of male reproductive disorders[J]. Toxicol Appl Pharmacol. 2005;207(2 Suppl):501-505.
42.Kaleva M, Virtanen HE, Haavisto AM,et al. Circannual rhythm in the incidence of cryptorchidism in Finland[J].Int J Androl. 2005;28(1):53-57.
43.Skakkebaek NE, J?rgensen N. Testicular dysgenesis and fertility[J]. Andrologia. 2005 ;37(6):217-218.
44.Lassurguère J, Livera G, Habert R, et al. Time- and dose-related effects of estradiol and diethylstilbestrol on the morphology and function of the fetal rat testis in culture[J]. Toxicol Sci. 2003;73(1):160-169.
45.Aravindakshan J, Paquet V, Gregory M,et al. Consequences of xenoestrogen exposure on male reproductive function in spottail shiners (Notropis hudsonius) [J]. Toxicol Sci. 2004;78(1):156-165.
46.Martinovi? D, Hogarth WT, Jones RE,et al. Environmental estrogens suppress hormones, behavior, and reproductive fitness in male fathead minnows[J]. Environ Toxicol Chem. 2007;26(2):271-8.
47.Doolan CM, Harvey BJ. A Galphas protein-coupled membrane receptor, distinct from the classical oestrogen receptor, transduces rapid effects of oestradiol on [Ca2+]i in female ratdistal colon[J]. Mol Cell Endocrinol. 2003;199(1-2):87-103.
48.Haynes MP, Li L, Sinha D,et al. Src kinase mediates phosphatidylinositol 3-kinase/Akt-dependent rapid endothelial nitric-oxide synthase activation by estrogen[J]. J Biol Chem. 2003;278(4):2118-2123.
49.Cunha GR, Forsberg JG, Golden R, et al. New approaches for estimating risk from exposure to diethylstilbestrol[J]. Environ Health Perspect. 1999 Aug;107 Suppl 4:625-30.
50.Bulayeva NN, Watson CS. Xenoestrogen-induced ERK-1 and ERK-2 activation via multiple membrane-initiated signaling pathways[J]. Environ Health Perspect. 2004;112(15):1481-1487.
51.Takeuchi A, Mishina Y, Miyaishi O,et al. Heterozygosity with respect to Zfp148 causes complete loss of fetal germ cells during mouse embryogenesis[J]. Nat Genet. 2003;33(2):172-176.
52.Maga G, Hubscher U. Proliferating cell nuclear antigen(PCNA), A dancer with many partners[J]. J Cell Sci, 2003;116(pt 15):3051-3060.
53.El Zoghbi CS, Favorito LA, Costa WS,et al. Structural analysis of gubernaculum testis in cryptorchid patients submitted to treatment with human chorionic gonadotrophin[J]. Int Braz J Urol. 2007;33(2):223-229.
54.Tanyel FC. The descent of testis and reason for failed descent[J]. Turk J Pediatr. 2004;46(Suppl):7-17.
55.Tanyel FC, Talim B, Atilla P,et al. Myogenesis within the human gubernaculum: histological and immunohistochemical evaluation[J]. Eur J Pediatr Surg. 2005;15(3):175-179.
56.蒋学武,邓汪东,胡显良等.外源性雌激素对小鼠睾丸引带收缩和增殖活性的影响[J].中华小儿外科杂志,2005;26(9):488-491.
57.Costa WS, Sampaio FJ, Favorito LA,et al. Testicular migration: remodeling of connective tissue and muscle cells in human gubernaculum testis[J]. J Urol. 2002;167(5):2171-2176.
58.Tomiyama H, Hutson JM. Contractility of rat gubernacula affected by calcitonin gene-related peptide and beta-agonist[J]. J Pediatr Surg. 2005;40(4):683-687.
59.Sasaki Y, Hutson JM, Clarke MC. The role of sympathetic innervation in the developing rat gubernaculum[J]. J Pediatr Surg. 2007;42(2):350-354.
1) Carlsen E, Giwercman A, Keiding N, et al. Evidence for decreasing quality of semen during past 50 years[J]. Brmed J.1992.305(6854):609-613.
2) Gerald R. Cunha,1 John-Gunnar Forsberg,2 Robert Golden,et al. New Approaches for Estimating Risk from Exposure to Diethyistilbestrol[J].Environment Health Perspectives,1999;107:625-630.
3) Herbst AL, Ulfelder H, Poskanzer DC. Adenocarcinoma of the vagina:Association of maternal stilbestrol therapy with tumor appearance in young women[J]. N Engl J Med. 1971,284(15):878-881.
4) Klip H, Verloop J, van Gool JD, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study[J]. Lancet,2002,359(9312):1102-1107.
5) Low-Dose Peer Review Panel(EPA,NIEHS and NIH-NTP).National Toxicology Proguam’s Report of the Endocine Disruptors Low-Dose Peer Review.2001,8,p53-54. http://ntp.niehs.nih.gov.
6) Suh RS, Wang X, Ohl DA, et al. Influence of Prenatal Diethylstilbestrol (DES) and dietary phyto- estrogens on sperm function and reproductive organ development in male mice[J]. Fertility and Sterility 2003;80(Suppl 3):235.
7) Newbold RR, Jefferson WN, Padilla-Banks E, et al. Developmental exposure to diethylstilbestrol (DES) alters uterine response to estrogens in prepubescent mice: low versus high dose effects[J]. Reproductive Toxicology,2004;18(3):399-406.
8) Emmen JM, McLuskey A, Adham IM, et al. Involvement of insulin-like factor 3 (Insl3) in diethylstilbestrol-induced cryptorchidism[J]. Endocrinology. 2000;141(2):846-9.
9)龚春雨,鲁刚,李虹,等.己烯雌酚建立尿道下裂大鼠模型的实验研究[J].中华小儿外科杂志,2004,25(2):180-181.
10) Inano H, Suzuki K, Onoda M, et al. Relationship between induction of mammary tumors and change of testicular functions in male rats following gamma-ray irradiation and/or diethylstilbestrol[J]. Carcinogenesis. 1996;17(2):355-360.
11) Troisi R, Hatch EE, Titus-Ernstoff L, et al. Cancer risk in women prenatally exposed to diethylstilbestrol[J]. Int J Cancer. 2007;121(2):356-360.
12) Green S, Walter P, Greene G, et al. Cloning of the human oestrogen receptor cDNA[J]. J Steroid Biochem. 1986;24(1):77-83.
13) Kuiper GG, Enmark E, Pelto-Huikko M, et al. Cloning of a novel receptor expressed in rat prostate and ovary[J]. Proc Natl Acad Sci U S A. 1996;93(12):5925-5930.
14) Levin ER. Cellular Functions of the Plasma Membrane Estrogen Receptor[J]. Trends Endocrinol Metab. 1999;10(9):374-377.
15) Adeoya-Osiguwa SA, Markoulaki S, Pocock V, et al. 17beta-Estradiol and environmental estrogens significantly affect mammalian sperm function[J]. Hum Reprod. 2003;18(1):100-107.
16) Zsarnovszky A, Le HH, Wang HS,et al. Ontogeny of rapid estrogen-mediated extracellular signal-regulated kinase signaling in the rat cerebellar cortex: potent nongenomic agonist and endocrine disrupting activity of the xenoestrogen bisphenol A[J]. Endocrinology. 2005;146(12):5388-5396.
17) Watson CS, Alyea RA, Jeng YJ,et al. Nongenomic actions of low concentration estrogens and xenoestrogens on multiple tissues[J]. Mol Cell Endocrinol. 2007;274(1-2):1-7.
18) Watson CS, Bulayeva NN, Wozniak AL,et al. Xenoestrogens are potent activators of nongenomic estrogenic responses[J]. Steroids. 2007;72(2):124-134.
19) Cowley SM, Hoare S, Mosselman S,et al. Estrogen receptors alpha and beta form heterodimers on DNA[J]. J Biol Chem. 1997;272(32):19858-19862.
20) Chen D, Washbrook E, Sarwar N,et al. Phosphorylation of human estrogen receptor alpha at serine 118 by two distinct signal transduction pathways revealed by phosphorylation-specific antisera[J]. Oncogene. 2002;21(32):4921-4931.
21) Saunders PT, Sharpe RM, Williams K,et al. Differential expression of oestrogen receptor alpha and beta proteins in the testes and male reproductive system of human and non-human primates[J]. Mol Hum Reprod. 2001;7(3):227-236.
22) Zhou Q, Nie R, Prins GS, et al. Localization of androgen and estrogen receptors in adult male mouse reproductive tract[J]. J Androl. 2002;23(6):870-881.
23) Bartlett JE, Washburn T, Eddy EM,et al. Early development of the gubernaculum and cremaster sac in estrogen receptor knockout mice[J]. Urol Res. 2001;29(3):163-167.
24) Staub C, Rauch M, Ferrière F,et al. Expression of estrogen receptor ESR1 and its 46-kDa variant in the gubernaculum testis[J]. Biol Reprod. 2005;73(4):703-712.
25) Cederroth CR, Schaad O, Descombes P, et al. Estrogen receptor alpha is a major contributorto estrogen-mediated fetal testis dysgenesis and cryptorchidism[J]. Endocrinology. 2007;148(11):5507-5519.
26) Guillette LJ Jr, Gross TS, Masson GR, Matter JM, et al. Developmental abnormalities of the gonad and abnormal sex hormone concentrations in juvenile alligators from contaminated and control lakes in Florida[J]. Environ Health Perspect. 1994;102(8):680-688.
27) Paris F, Jeandel C, Servant N, et al. Increased serum estrogenic bioactivity in three male newborns with ambiguous genitalia: a potential consequence of prenatal exposure to environmental endocrine disruptors[J]. Environ Res, 2006;100(1):39-43.
28)王金平,王滨有.尿道下裂危险因素的病例对照研究[J].中华流行病学杂志,2004,25(3):261-264.
29) Pierik FH, Burdorf A, Deddens JA, et al. Maternal and paternal risk factors for cryptorchidism and hypospadias: a case-control study in newborn boys[J]. Environ Health Perspect. 2004,112(15):1570-1576.
30) Shono T, Suita S, Kai H, et al. The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats[J]. J Pediatr Surg, 2004,39(2):213-216.
31) Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat[J]. Toxicol Sci,2000,58(2):350-365.
32) Virtanen HE, Rajpert-De Meyts E, Main KM,et al. Testicular dysgenesis syndrome and the development and occurrence of male reproductive disorders[J]. Toxicol Appl Pharmacol. 2005;207(2 Suppl):501-505.
33) Kaleva M, Virtanen HE, Haavisto AM,et al. Circannual rhythm in the incidence of cryptorchidism in Finland[J]. Int J Androl. 2005;28(1):53-57.
34) Skakkebaek NE, J?rgensen N. Testicular dysgenesis and fertility[J]. Andrologia. 2005 ;37(6):217-218.
35) Lassurguère J, Livera G, Habert R, et al. Time- and dose-related effects of estradiol and diethylstilbestrol on the morphology and function of the fetal rat testis in culture[J]. Toxicol Sci. 2003;73(1):160-169.
36) Aravindakshan J, Paquet V, Gregory M,et al. Consequences of xenoestrogen exposure onmale reproductive function in spottail shiners (Notropis hudsonius) [J]. Toxicol Sci. 2004;78(1):156-165.
37) Martinovi? D, Hogarth WT, Jones RE,et al. Environmental estrogens suppress hormones, behavior, and reproductive fitness in male fathead minnows[J]. Environ Toxicol Chem. 2007;26(2):271-8.
38) McKinnll C, Sharpe RM, Mahood K, et al. Expression of insulin-like factor 3 protein in the rat testis during fetal and postnatal development and in relation to cryptorchidism induced by in utero exposure to di (n-Butyl) phthalate[J]. Endocrinology. 2005;146(10):4536-44.
39) Christophe Staub, Michel Rauch,Franc,et al. Expression of Estrogen Receptor ESR1 and Its 46-kDa Variant in the Gubernaculum Testis[J]. BIOLOGY OF REPRODUCTION, 2005;73:703–712.
40) Doolan CM, Harvey BJ. A Galphas protein-coupled membrane receptor, distinct from the classical oestrogen receptor, transduces rapid effects of oestradiol on [Ca2+]i in female rat distal colon[J]. Mol Cell Endocrinol. 2003;199(1-2):87-103.
41) Haynes MP, Li L, Sinha D,et al. Src kinase mediates phosphatidylinositol 3-kinase/Akt-dependent rapid endothelial nitric-oxide synthase activation by estrogen[J]. J Biol Chem. 2003;278(4):2118-2123.
42) Fraser LR, Beyret E, Milligan SR,et al. Effects of estrogenic xenobiotics on human and mouse spermatozoa[J]. Hum Reprod. 2006;21(5):1184-1193.
43) Brucker-Davis F, Thayer K, Colborn T. Significant effects of mild endogenous hormonal changes in humans: considerations for low-dose testing[J]. Environ Health Perspect. 2001;109(Suppl 1):21-26.
44) Bulayeva NN, Watson CS. Xenoestrogen-induced ERK-1 and ERK-2 activation via multiple membrane-initiated signaling pathways[J]. Environ Health Perspect. 2004;112(15):1481-1487.
[1]王光义,陈绍基,蒋学武,等.尿道下裂伴阴茎发育不良的内分泌改变及治疗[J].中华小儿外科杂志,1995;16(1):23-25.
[2]吴艳乔,王艳萍,朱军,等.国人尿道下裂的流行病学特征[J].中华泌尿外科杂志,1997,18(12):753-757.
[3]吴艳乔,代礼,王艳萍,等.中国儿童尿道下裂发生率的变化趋势[J].四川大学学报:医学版,2005,36(2):274-276.
[4]张伟,张波,邓兵.尿道下裂患儿性染色体异常的细胞及分子遗传学研究[J].中华实用医学,2004,6(16):4-6.
[5] Chu JH, Zhang RM, Zhao ZM, et al. Male Fertility Is Compatible with an Arg840Cys Substitution in the AR in a Large Chinese Family Affected with Divergent Phenotypes of AR Insensitivity Syndrome[J]. J Clin Endocrinol Metab,2002,87(1):347–351.
[6] Abbate B, Danti DA, Mattei R, et al. Genetic aspects of hypospadias: considerations apropos of 362 cases[J]. Pediatr Med Chir,1994,16(1):77-79.
[7] Kim KS, Liu W, Cunha GR, et al. Expression of the androgen receptor and 5alpha-reductase type 2 in the developing human fetal penis and urethra[J]. Cell Tissue Res,2002,307(2):145-153.
[8]贾悦,王晓东,崔毓桂,等. 5α-还原酶在男性生殖中的作用[J].生殖医学杂志,2003,12(2):115-118.
[9] Vilchis F, Mendez JP, Canto P,et al. Identification of missense mutations in the SRD5A2 gene from patients with steroid 5alpha-reductase 2 deficiency[J]. Clin Endocrinol (Oxf),2000 ,52(3):383-387.
[10] Silver R I, RussellDW. 5 alpha-reductase type 2mutations are present in some boys with isolated hypospadias[J]. J Urol ,1999, 162 (3 Pt2 ): 1142-1145.
[11]周李,梅骅,刘唐彬,等.先天性尿道下裂与SRD5A2及SRY基因突变关系研究[J].中华医学遗传学杂志,1999,16(5);311-314.
[12] Wang YP, Li Q, Xu JJ, et al. Mutation analysis of five candidate genes in Chinese patients with hypospadias[J]. Eur J Hum Genet ,2004,12(9):706-712.
[13]徐家杰,李森恺,李强,等.尿道下裂患者SRD5A2基因突变的研究[J].中华整形外科杂志,2006,22(2): 139-141.
[14] Aaronson IA, Cakmak MA, Key LL. Defects of the testosterone biosynthetic pathway in boys with hypospadias[J]. J Urol ,1997,157(5):1884-1888.
[15]唐建华,袁继炎.尿道下裂与Ⅱ型3β-羟类固醇脱氢酶基因突变的研究[J].临床小儿外科杂志,2002,1(4):282-285.
[16]李春,谢家伦,卢汉平.先天性尿道下裂外生殖器皮肤细胞内雄激素受体的含量测定[J].衡阳医学院学报, 1998,26(2):179-181.
[17]李强,李森恺,徐家杰,等.尿道下裂患者雄激素受体基因突变的研究[J].中华整形外科杂志,2004,20(6):421-424.
[18]徐哲,郑克立,乔慧,等.雄激素受体基因(CAG)n重复多态与先天性尿道下裂的关系[J].临床小儿外科杂志,2002,1(6):435-437.
[19] Aschim EL, Nordenskjold A, Giwercman A, et al. Linkage between cryptorchidism, hypospadias, and GGN repeat length in the androgen receptor gene[J]. J Clin Endocrinol Metab ,2004,89(10):5105-5109.
[20]蒋学武,陈中献,陈绍基,等.先天性尿道下裂患儿阴茎组织中多种性激素受体的比较研究[J].中国男科学杂志,2000,14 (1) :16-18.
[21] Castineyra G, Copelli S, Levalle O. 46, XX male: clinical, hormonal/genetic findings[J]. Arch Androl ,2002,48 (4) : 251-257.
[22] Quigley DI, McDonald MT, Krishnamuthy V,et al. Triploid mosaicism in a 45,X/69,XXY infant[J]. Am J Med Genet A. 2005, 138 ( 2 ) : 171-4.
[23]刘国庆,赵蕊,叶志纯,等.先天性尿道下裂与SRY基因关系的探讨[J].中华泌尿外科志,2001,22 (4) : 241-242.
[24]李强,李森恺,徐家杰.尿道下裂病人中SRY基因缺失的快速检测[J].中华整形外科杂志,2002 (S1) : 40-41.
[25] Moreno-Garcia M, Miranda EB. Chromosomal anomalies in cryptorchidism and hypospadias[J]. J Urol, 2002 ,168 ( 5 ) : 2170-2172.
[26]游慧灵,余敏. 14例尿道下裂患儿的细胞遗传学分析[J].中国优生与遗传志,2001,9(3):54,127.
[27] Paris F, Jeandel C, Servant N, et al. Increased serum estrogenic bioactivity in three male newborns with ambiguous genitalia: a potential consequence of prenatal exposure to environmental endocrine disruptors[J]. Environ Res, 2006,100 ( 1 ) : 39-43.
[28]罗艳,栾荣生,朱军,等.尿道下裂与环境污染的关系—Poisson回归模型与GM (1 , N)模型结合分析.现代预防医学[J], 2005 , 32 ( 1 ) : 1-2 , 15.
[29] Kun Suk Kim, Carlos R. Torres Jr., et al. Induction of hypospadias in a murine model by maternal exposure to synthetic estrogens[J]. J Environmental Research ,2004, 94 : 267–275.
[30] Willingham E, Agras K, Vilela M, et al. Loratadine Exerts Estrogen-Like Effects and Disrupts Penile Development in the Mouse[J]. J Urol, 2006,175 (2) :723-726.
[31]龚春雨,鲁刚,李虹,等.己烯雌酚建立尿道下裂大鼠模型的实验研究[J].中华小儿外科杂志,2004,25 (2) :180-181.
[32] Klip H, Verloop J, van Gool JD, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero: a cohort study[J]. Lancet , 2002,359 (9312) : 1102-1107.
[33] Kallen B. Use of antihistamine drugs in early pregnancy and delivery outcome[J]. J Matern Fetal Neonatal Med , 2002 ,11 (3) : 146-152.
[34] Brouwers MM, Feitz WF, Roelofs LA, et al. Hypospadias: a transgenerational effect of diethylstilbestrol[J] Hum Reprod, 2006,21 ( 3 ) : 666-669.
[35] Palmer JR, Wise LA, Robboy SJ, et al. Hypospadias in sons of women exposed to diethylstilbestrol in utero[J]. Epidemiology , 2005 ,16 ( 4 ) : 583-586.
[36] Centers for Disease Control and Prevention (CDC). Evaluation of an association between loratadine and hypospadias--United States, 1997-2001. MMWR Morb Mortal Wkly Rep, 2004,53(10):219-21.
[37]贺厚光,张炜,朱佳庚.小鼠尿道下裂动物模型的建立[J].中华男科学,2004,10 ( 3 ) : 172-174.
[38]韩雪冰,杨晓峰,任连生,等.先天性尿道下裂动物模型的建立[J].中华实验外科学杂志,2005, 22 ( 5 ) : 590.
[39]李岩,张浩,王文冬,等.小剂量氟他胺对大鼠性分化影响的实验研究[J].四川大学学报(医学版) , 2006 ,37 ( 2 ) : 258-261.
[40]张炜,袁琳,吴婷,等.邻苯二甲酸二丁酯诱导尿道下裂大鼠模型的建立及其作用机制[J].中华实验外科杂志, 2005 , 22 ( 2 ) : 246-248.
[41] Shono T, Suita S, Kai H, et al. The effect of a prenatal androgen disruptor, vinclozolin, on gubernacular migration and testicular descent in rats[J]. J Pediatr Surg, 2004, 39 ( 2) : 213-216.
[42] Gray LE Jr, Ostby J, Furr J, et al. Perinatal exposure to the phthalates DEHP, BBP, and DINP, but not DEP, DMP, or DOTP, alters sexual differentiation of the male rat[J]. Toxicol Sci , 2000 , 58 ( 2 ) : 350-365.
[43]王金平,王滨有.尿道下裂危险因素的病例对照研究[J].中华流行病学杂志, 2004 , 25 ( 3 ) : 261-264.
[44] Pierik FH, Burdorf A, Deddens JA, et al. Maternal and paternal risk factors for cryptorchidism and hypospadias: a case-control study in newborn boys[J]. Environ Health Perspect. 2004 , 112 ( 15 ) : 1570-1576.
[45] North K, Golding J. A maternal vegetarian diet in pregnancy is associated with hypospadias. The ALSPAC Study Team. Avon Longitudinal Study of Pregnancy and Childhood[J]. BJU Int. 2000 , 85 (1) :107-113.
[46] LIU BC, WANG Z, LIN GT, et al. Activating Transcription Factor 3 Is Up-Regulated in Patients with Hypospadias[J]. Pediatric Research , 2005 ,58( 6) : 1280-1283 .
[47] Carmichael SL, Shaw GM, Laurent C, et al . Maternal progestin intake and risk of hypospadias[J]. Arch Pediatr Adolesc Med , 2005 , 159 ( 10 ) : 957-62.
[48] Silver RI, Rodriguez R, Chang TS, et a l. In vitro Fertilization is assosiated with an increased risk of hypospadias[J]. J Uro l, 1999 , 161 : 1954-1957.
[49]王军,袁继炎,周学锋.尿道下裂患儿包皮组织中表皮生长因子及其受体的表达及意义[J].中华小儿外科杂志, 2001 , 22 ( 1 ) : 30-31.
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