Di-n-butyl phthalate (DBP) is an environmental endocrine disruptor with developmental nephrotoxicity, but its long-term effects on renal fibrotic remodeling in offspring remain incompletely understood. In this study, a maternal DBP exposure model was established in pregnant Sprague-Dawley rats (750 mg/kg/day, GD14-GD18), and adult male offspring were examined at postnatal day 42. DBP-exposed offspring showed intrauterine growth restriction, reduced relative kidney weight, renal collagen deposition, and epithelial-mesenchymal transition (EMT)-related molecular changes. Transcriptomic analysis identified hemoglobin subunit beta (HBB) as a prominently downregulated gene in renal tissues of DBP-exposed offspring. HBB expression was reduced by approximately four-fold and was accompanied by increased oxidative stress, including elevated reactive oxygen species and malondialdehyde levels and decreased total superoxide dismutase activity. DBP exposure was also associated with increased expression of NRF2 and its downstream antioxidant targets HO-1 and NQO1. In HK-2 cells, HBB overexpression attenuated DBP-induced oxidative stress and EMT-related marker changes, while N-acetylcysteine (NAC) reduced oxidative stress and restored HBB and EMT-related protein expression. In adult male offspring, renal AAV-HBB delivery partially restored HBB expression and reduced DBP-associated collagen deposition, oxidative stress, and EMT-related changes. These findings indicate that maternal DBP exposure is associated with adult renal fibrotic remodeling in male offspring, with HBB downregulation, oxidative stress, NRF2-related antioxidant responses, and EMT-related alterations as key molecular features. HBB may serve as a candidate biomarker and intervention target for DBP-associated developmental nephrotoxicity, pending further validation.

Maternal exposure to di-n-butyl phthalate (DBP) has been linked to the induction of hypospadias; however, the underlying mechanism remains unclear. Necroptosis is reported to be implicated in developmental malformations. This study aimed to investigate the underlying mechanism of necroptosis in the development of hypospadias. DBP was dissolved in corn oil, and pregnant rats were administered a precisely measured dose of DBP (750 mg/kg/day) via gastric intubation from gestation day 14-18. Control rats received only corn oil. The day of birth was considered postnatal day (PND) 1. Male hypospadias rats were identified on PND 7. Genital tubercle tissues were collected and stored at -80°C for subsequent PCR analysis, cryopreserved in liquid nitrogen for western blot, or fixed in formalin for immunohistochemistry (IHC) staining. IHC staining and western blot analysis revealed increased expression of RIPK1 and necroptosis markers in genital tubercle (GT) tissue compared to the control group. Additionally, higher levels of EMT and impaired androgen receptor expression were observed in GT tissue. Exposure to increased DBP concentrations in rat primary uroepithelial cells (PUCs) led to elevated ROS production. Necroptosis markers and EMT expression levels were upregulated in PUCs following DBP incubation. Notably, treatment with DBP combined with necrostatin-1, a necroptosis inhibitor, reduced the expression of EMT markers and ROS production compared to DBP treatment alone. In vitro studies further revealed that DBP-induced necroptosis promoted the degradation of E-cadherin through the ubiquitin-proteasome pathway in PUCs. Our findings suggest that maternal exposure to DBP promotes necroptosis in uroepithelial cells by elevating ROS level and EMT status. Thus, necroptosis may play an essential role in the development of hypospadias.

Maternal exposure to dibutyl phthalate (DBP) induces renal fibrosis in offspring. However, the specific roles of connexin 43 (Cx43) in DBP-induced renal fibrosis remain unknown. Therefore, in this study, we analysed the expression of Cx43 in renal tubular epithelial cells (RTECs) with or without DBP exposure using reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and western blotting. A small interfering RNA against Cx43 was introduced to assess its role in epithelial–mesenchymal transition (EMT) of RTECs caused by 100 μmol/L DBP. Bioinformatics analysis was conducted with AMP-activated protein kinase (AMPK)-α2 and angiotensin (Ang) II inhibitors to determine the mechanisms involved in the expression of Cx43 in HK-2 cells. RT-qPCR and western blotting revealed that DBP increased the expression of Cx43 in vitro. Moreover, Cx43 knockdown significantly alleviated DBP-induced EMT caused by DBP in HK-2 cells. Bioinformatics analysis with AMPKα2 and Ang II inhibitors revealed that DBP upregulated Cx43 expression by activating the Ang II/AMPKα2 signaling pathway. Our findings indicate that DBP induces renal fibrosis by activating Ang II/AMPKα2/Cx43 signaling pathway and EMT in RETCs, suggesting a potential target for the treatment of renal fibrosis.

BackgroundBladder cancer is a common malignant type in the world, and over 90% are transitional cell carcinoma. While the impact of inflammatory response on cancer progression has been reported, the role of inflammatory response-associated genes (IRAGs) in transitional bladder cancer still needs to be understood.MethodsIn this study, IRAGs were download from Molecular Signature Database (MSigDB). The transcriptional expression and matched clinicopathological data were separately obtained from public databases. The TCGA-BLCA cohort was used to identify the differentially expressed IRAGs, and prognostic IRAGs were filtrated by univariate survival analysis. The intersection between them was displayed by Venn diagram. Based on least absolute shrinkage and selection operator (LASSO) regression analysis method, the TCGA-BLCA cohort was used to construct a risk signature. Survival analysis was conducted to calculate the overall survival (OS) in TCGA and GSE13507 cohort between two groups. We then conducted univariate and multivariate survival analyses to identify independently significant indicators for prognosis. Relationships between the risk scores and age, grade, stage, immune cell infiltration, immune function, and drug sensitivity were demonstrated by correlation analysis. The expression level of prognostic genes in vivo and in vitro were determined by qRT-PCR assay.ResultsComparing with normal tissues, there were 49 differentially expressed IRAGs in cancer tissues, and 12 of them were markedly related to the prognosis in TCGA cohort for transitional bladder cancer patients. Based on LASSO regression analysis, a risk model consists of 10 IRAGs was established. Comparing with high-risk groups, survival analysis showed that patients in low-risk groups were more likely to have a better survival time in TCGA and GSE13507 cohorts. Besides, the accuracy of the model in predicting prognosis is acceptable, which is demonstrated by receiver operating characteristic curve (ROC) analysis. Age, stage, and risk scores variables were identified as the independently significant indicators for survival in transitional bladder cancer. Correlation analysis represented that the risk score was identified to be significantly related to the above variables except gender variable. Moreover, the expression level of prognostic genes in vivo and in vitro was markedly upregulated for transitional bladder cancer.ConclusionsA novel model based on the 10 IRAGs that can be used to predict survival time for transitional bladder cancer. In addition, this study may provide treatment strategies according to the drug sensitivity in the future.

Objective: This study focused on the oxidative stress effect of di-n-butyl phthalate (DBP) on development of the urinary system. Methods: We examined the mRNA expression of genital tubercle (GT) in control and DBP induced hypospadias group by Affymetrix Rat 230 2.0 Array. Real-time PCR and Western Blot were used to detect the protein and mRNA expression levels of inositol-1,4,5-triphate-receptor (IP3R) and epithelial-mesenchymal-transition (EMT)-related molecular markers, such as E-cadherin, beta-Catenin, Snail, N-cadherin, in the GT of hypospadiac male rats and controls. The results of array were further confirmed in vitro. The changes of intracellular calcium concentration in urethral epithelial cells were detected by Fluo-3-AM before and after DBP treatment. The levels of reactive oxygen species (ROS) in urethral epithelial cells were measured by DCFH-DA with different concentrations of DBP (0, 1, 10, 100 mu mol/L) treatment. Results: The mRNA expression profiles of GT in control and DBP induced hypospadias group showed high expression of IP3R and the abnormalities of EMT. Compared to the control group, the expression levels of IP3R, E-cadherin and beta-Catenin increased at both the protein and mRNA levels. However the expression levels of Snail and N-cadherin decreased. The intracellular calcium concentration increased significantly after DBP treatment. The effect of DBP on urethral epithelial cells was linked to the generation of oxidative stress. Conclusion: DBP can influence the development of GT through its oxidative stress effect, which significantly increases the concentration of calcium and inhibits EMT in urethral epithelial cells, and block the fusion process of urethral groove, causing the occurrence of hypospadias. This study provides a new understanding of DBP's molecular mechanisms on hypospadias and may lead to new treatment strategies for the disease.

The study focused on the toxicological effect of Di-n-butyl phthalate (DBP) on the expression of Phosphorylated signal transducer and activator of transcription 1 (pSTAT1) -regulated Forkhead box protein M1 (FoxM1), which might provide a new understanding of gestational diabetes mellitus (GDM) development and a potential target for treatment. Streptozotocin (STZ) (40 mg/kg) was introduced in maternal rats by intraperitoneal injection on gestation day 0 (GD 0) in the STZ and STZ + DBP groups. DBP was introduced in maternal rats by oral feeding in the STZ + DBP group over the following 3 days (750 mg/kg/day). The changes in fasting blood glucose level in rats were detected on GD 1 and GD 5. The insulin levels in maternal rats and PIBCs were measured on GD 18. The Oral Glucose Tolerance Test (OGTT) test was performed on GD 18 to check the stability of the GDM model. The primary islet β cells (PIBCs) were established for in vitro experiments. We examined the FoxM1 and pSTAT1 expression in pancreas by immunohistochemistry. Real-time PCR and Western blot were used to detect the pSTAR1 and FoxM1 protein and mRNA gene expression levels in PIBCs. Cell Counting Kit-8 (CCK-8) and flow cytometric analysis was used to test the viability and apoptosis of cells. The results showed that the STZ + DBP group had higher glucose and lower insulin secretion levels than the other groups by both fasting test and OGTT. FoxM1 was significantly suppressed while pSTAT1 was highly expressed after DBP exposure. FoxM1 could be regulated by pSTAT1. DBP can influence the progression of GDM through its toxicological effect, which significantly increases the expression of pSTAT1 and suppresses FoxM1, causing a decline in β cell viability.

目的:验证邻苯二甲酸二丁酯(DBP)是通过降低血清雄激素水平导致自噬异常激活,同时探讨DBP致子代大鼠尿道下裂发生的具体机制.方法:将孕鼠随机分为DBP染毒组与对照组,并于妊娠期14-18天通过灌胃的方式,分别用DBP(750 mg/kg/天)饲养DBP染毒,用等量花生油饲养对照组.依照此方法成功构建了子代新生大鼠尿道下裂模型.采集子鼠生殖结节(GT)用福尔马林保存,用免疫组织化学(IHC)染色观察生殖结节组织中自噬水平,即LC3B及Beclin1表达水平;在子鼠麻醉后采集血液标本,用放射免疫分析方法观测子鼠血清睾酮水平.在原代大鼠尿路上皮细胞(PUECs)基础上,用Western印迹方法检测有无双氢睾酮(DHT)对PUECs中LC3I、LC3Ⅱ及Beclin1表达水平影响.结果:DBP染毒组尿道下裂发生率为42.3％,对照组子代无尿道下裂.DBP染毒组子代GT组织中自噬表达较对照组明显增加.DBP染毒组(n=10)较对照组中血清睾酮水平有明显差异(n=10)(P＜0.05).体外研究表明DHT缺乏组Beclin1及LC3蛋白转化率水平较对照组升高.结论:孕期暴露于DBP可以诱发子代尿道下裂发生,这可能是由于DBP降低子鼠雄激素水平促使自噬发生导致的,然而该疾病的机制仍需要进一步研究.

目的:探讨基于团队的学习模式(Team-Based Learning,TBL)和基于讲座的学习模式(Lecture-Based Learning,LBL)在泌尿外科临床实习教学实践中的应用及效果评价.方法:选取2017年6月至2018年3月在上海交通大学附属第一人民医院泌尿外科进行临床实习的临床医学专业本科学生56名,按照成绩排序并逐次分配至实验组(28名)和对照组(28名).实验组采用TBL教学法,对照组采用LBL教学法课程结束后,对比分析两组的理论知识和实践能力得分,并采用调查问卷评价两组教学效果.结果:TBL组理论知识和临床实践考核成绩优于LBL组,TBL组对教学效果评价的满意度高于LBL组.结论:TBL教学法在理论知识学习和临床实践能力方面均有明显优势,调查问卷发现大部分学生对TBL教学法比较认可,但其结果还需要更大规模的研究验证.

肺真菌感染常发生于器官移植受者、恶性肿瘤患者、恶性血液病患者等免疫功能低下者,其主要由隐球菌、念珠菌、曲霉菌等真菌侵入肺部导致.但随着城市化发展与人群流动的日益频繁,免疫健全者肺部自然感染隐球菌的可能性也大幅度提高[1].伏立康唑(voriconazole)是第2代三唑类广谱抗真菌药物,作用机制是通过抑制真菌中由CYP450介导的14α-甾醇去甲基化,使真菌细胞膜重要成分麦角甾醇的生物合成受阻,而产生抗真菌作用,用于治疗严重侵袭性真菌感染[2,3].

Maternal exposure to di-n-butyl phthalate (DBP) induces hypospadias, but the underlying mechanisms remain elusive. Here we hypothesize that aberrant activation of autophagy and epithelial-mesenchymal transition (EMT) are the leading cause of DBP-related hypospadias. Pregnant rats received DBP orally at a dose of 750 mg/kg/day during gestational days 14-18. In DBP-induced hypospadiac male offspring, immunohistochemistry (IHC) staining and Western blot showed increased expression of autophagy and EMT markers in genital tubercle (GT) tissue compared to the control. In addition, lower testosterone levels and androgen receptor (AR) expression in GT tissue were detected. In vitro studies revealed that impaired AR signaling was involved in DBP-induced autophagy and autophagy activation furthermore promoted EMT in urethral epithelial cells. DBP combined with chloroquine, an autophagy inhibitor, reduced the expression of EMT markers compared with DBP treatment alone, while DBP combined with the autophagy inducer rapamycin elevated the expression of EMT markers. The autophagy-lysosomal pathway inhibitor CQ but not proteasome inhibitor MG-132 rescued the decrease of E-cadherin after DBP treatment, which indicated autophagy-induced E-cadherin degradation contributes to DBP-related EMT. Taken together, our findings show that prenatal exposure to DBP induces abnormal autophagy and EMT that may play important roles in hypospadias development.

BACKGROUND:We previously demonstrated that maternal exposure to di-n-butyl phthalate (DBP) induces dysplasia of the kidney in newborn male offspring and renal fibrosis in adults. But the underlying mechanisms remain elusive. Fgf10/Fgfr2 and androgen receptor (AR) are known to be important for renal development. We therefore investigated whether these genes are involved in DBP-induced renal fibrosis. MATERIALS AND METHODS:Using Sprague-Dawley rats and rat renal proximal tubular cells (NRK52E), we determined the potential involvement of Fgf10, Fgfr2 and AR in DBP-induced renal fibrosis. RESULTS:We found that maternal exposure to DBP induces renal fibrosis in adult male offspring. A lower serum testosterone concentration and reduced expression of Fgf10, Fgfr2 and AR were detected in these animals. These was a trend toward lower expression of Fgf10, Fgfr2 and AR in NRK52E cells subjected to DBP exposure. Furthermore, higher expression levels of TGF-β and α-SMA were observed in abnormal renal tissue and DBP-treated NRK52E cells. CONCLUSION:Our findings suggest the potential involvement of Fgf10/Fgfr2 and AR in renal fibrosis of adult male rat offspring induced by prenatal exposure to DBP. The anti-androgenic effects of DBP might play an important role in this pathological process.

This study was to determine the impact of maternal exposure to di-n-butyl phthalate (DBP) on renal development and fibrosis in adult offspring. Pregnant rats received DBP at a dose of 850 mg/kg BW/day by oral perfusion during gestational days 14-18. In DBP exposed newborn offspring, gross observation and histopathological examination revealed the dysplasia of kidney. The expression of genes related to renal development was also changed. In DBP exposed adult offspring, histopathological examination and Masson's trichrome staining revealed the pathological changes of renal fibrosis. Furthermore, higher expression levels of transforming growth factor- β (TGF-β) and alpha-smooth muscle actin (α-SMA) were also detected. In vitro studies reveal that DBP promoted the activation of NRK49F cells and G2/M arrest in NRK52E cells at a sublethal dose. The effect of DBP on these cell lines was linked to the generation of oxidative stress. In addition, DBP induced oxidative stress in both renal fibroblasts and tubular epithelial cells, whereas vitamin C ameliorated the changes caused by DBP. In conclusion, our results showed that prenatal exposure to DBP may generate oxidative stress in both renal fibroblasts and tubular epithelial cells, leading to kidney dysplasia and renal fibrosis.

Objective To induce anorectal malformations (ARMs) in male rats by maternal exposure to di-n-butyl phthalate (DBP) during late pregnancy and examine the genetic characterization and morphological abnormalities in male offspring.Methods Twenty timed pregnant rats were randomly divided into two groups.DBP was administered by gastric intubation at a dose of 0,850 mg/kg/d from gestation day (GD) 12 to 18.On postnatal day (PND) 1,the incidence of ARMs was evaluated and body weight and anogenital distance (AGD) were measured.Themorphological abnormalities of ARMs were observed by histology.The mRNA expressions of such genes as Shh,Gli2,Gli3,Bmp4,Wnt5a,Hoxa13,Hoxd13,Fgf10 and Fgfr2 associated with anorectal development were examined by real-time polymerase chain reaction (PCR) in terminal rectum of ARM male offspring.Results The incidence of ARMs was 39.5％.Body weight and AGD of ARM male rats on PND1 were signi cantly lower than those of control (P＜0.05).The presence of ARMs was confirmed histologically.Remarkably decreased mRNA expressions of the above genes were observed in terminal rectum of ARM male offspring (P＜0.05).Conclusions During late pregnancy,an exposure to DBP at a dose of 850 mg/kg/d may lead to a high incidence of ARMs and developmental damage in male rats.And the down-regulations of the relevant genes may disturb the signaling networks in terminal rectum of ARMs.

目的：：探析黄金微针射频结合表皮生长因子在面部皮肤年轻化治疗中的应用效果.方法：选取2015年5月到2016年3月我院收治的面部皮肤老化30例作为对照组，给予光子嫩肤治疗；选取同期同类型30例作为研究组，给予黄金微针射频结合表皮生长因子治疗.对两组患者的临床治疗效果进行分析和对比.结果：给予两种不同的治疗方法后，研究组治疗总体有效率以及总体满意率均高于对照组(P＜0．05)．讨论：黄金微针射频结合表皮生长因子在面部皮肤年轻化治疗中具有极高的应用价值，值得大力推广以及应用.

This study was to compare the alterations of androgen cascades in di-n-butyl phthalate (DBP)-exposed male offspring without hypospadias (undeformed) versus those with hypospadias. To induce hypospadias in male offspring, pregnant rats received DBP via oral gavage at a dose of 750mg/kg BW/day during gestational days 14–18. The mRNA expression levels of genes downstream of the androgen signaling pathway, such as androgen receptor (AR) and Srd5a2, in testes of undeformed rat pups were similar to those in controls; in hypospadiac rat pups these levels were significantly lower than those of control pups. In contrast, both undeformed and hypospadiac rats had decreased serum testosterone levels, reduced mRNA expression of key enzymes in the androgen synthetic pathway in the testes, and ablated genes of developmental pathways, such as Shh, Bmp4, Fgf8, Fgf10 and Fgfr2, in the genital tubercle (GT) as compared to those in DBP-unexposed controls, albeit hypospadiac rats had a more severe decrement than those of undeformed rats. Although other possibilities cannot be excluded, our findings suggest that the relatively normal levels of testosterone-AR-Srd5a2 may contribute to the resistance to DBP toxicity in undeformed rats. In conclusion, our results showed a potential correlation between decreased testosterone levels, reduced mRNA expression of AR and Srd5a2 and the occurrence of hypospadias in male rat offspring prenatally exposed to DBP.

Anorectal malformations in combination with hypospadias (ARMs & hypospadias) are a type of complex congenital malformations. The underlying mechanisms of this deformity are largely unknown. In this study, we comprehensively characterized the dysplasia, histological malformations, and genetic changes of ARMs & hypospadias in male rats after maternal exposure to di-n-butyl phthalate (DBP) by gastric intubation at doses of 850mg/kg bw/day during GD11-15. On postnatal day 1, anatomical and histopathological analysis confirmed combined malformations of the genital tubercle (GT), terminal rectum (TR) and testes. DBP-induced dysplasia was also seen in the kidney, lung, spleen, heart and liver of ARMs & hypospadias male rats. Moreover, decreased levels of serum testosterone, as well as reduced expression of genes related to the androgen signaling pathway (Cyp11a1, Hsd3b, Scarb1, Star, AR, Srd5a2) were found in the testes of ARMs & hypospadias male rats after DBP exposure as compared to untreated controls. Further, decreased mRNA levels of Shh, Fgf10, Gli2, Gli3, Bmp4, Wnt5a, Hoxa13, Hoxd13, Fgfr2 and AR were observed in TR and GT in the ARMs & hypospadias group. These results provide evidence that prenatal exposure to DBP can lead to combined anorectal and urogenital malformations as well as dysplasia of the testes.

目的：观察干扰素-α联合窄谱中波紫外线治疗皮肤 T 细胞淋巴瘤的临床疗效。方法我院2012年1月至2013年12月期间所收治的皮肤 T 细胞淋巴瘤患者中选取20例作为此次研究对象，随机分为观察组和对照组，两组患者的年龄、性别等一般情况比较没有明显的差异，有可比性，P>0.05，具有统计学意义。结果观察组患者的治疗有效率为90.0%，对照组患者的治疗有效率为60.0%，观察组患者治疗有效率明显高于对照组患者，P<0.05，差异具有统计学意义。结论对于皮肤 T 细胞淋巴瘤患者，采用干扰素-α联合窄谱中波紫外线治疗，有助于提高患者治疗总有效率，安全性较高，具有一定的应用价值和治疗优势，值得大力推广使用。

The aim of this study is to compare the clinical efficacy and safety of retroperitoneal laparoscopic ureterolithotomy (RPLU) and ureteroscopic holmium laser lithotripsy (UHLL) as two minimally invasive procedures in managing obstructive upper ureteral calculi with concurrent urinary tract infections (UTI). The retrospective study included 189 patients who underwent unilateral obstructive upper ureteral stones with concurrent UTI from January 2007 to November 2014 at our institution. Patients received RPLU (81 cases) or UHLL (108 cases). All patients received preoperative anti-infection treatment (indwelling ureteral stent and/or preoperative antibiotics). Collected data, including sex, age, stone size, success rate, operation duration, post-operation hospitalization time, and post-operation complications, were compared. All patients were followed up for more than 6 months after surgeries, and no ureterostenosis occurred. The study included 189 patients, 41 (21.7 %) females and 148 (78.3 %) males with a medium age of 52 years (range 22–81 years). All surgeries were successfully performed without conversion to open surgery. Stone size in the RPLU group was larger than that of the UHLL group (16.1 ± 1.4 vs. 10.4 ± 1.6 mm, P = 0.012). Operative duration (P = 0.009) and hospitalization time (P < 0.001) in the UHLL group were significantly shorter than those in the RPLU group, whereas stone clearance rate was significantly higher in the RPLU group (100 vs. 88.9 %, P = 0.002). Of note, postoperative fever was more common in patients treated with UHLL (15 cases) versus RPLU (4 cases) (13.9 vs. 4.9 %, P = 0.043). Moreover, in the UHLL group, three patients without a preoperative indwelling ureteral stent were complicated with sepsis, which was not seen in RPLU group. In our study, the safety and stone clearance rate of RPLU are better than those of UHLL in the treatment of unilateral upper ureteric calculi with concurrent UTI. Preoperative antibiotics and indwelling ureteral stent may reduce the risk of postoperative infections.

Previous study have demonstrated that not only the anorectal development but also the general conditions of anorectal malformations (ARMs) male rats are severely affected by di-n-butyl phthalate (DBP) maternal exposure. However, the mechanisms underlying DBP-induced congenital defects remain elusive. Reportedly, Fgf10/Fgfr2 and androgen receptor (AR) are pivotal for the development of multiple organs. In this study, we therefore investigated the expression of Fgf10/Fgfr2 together with AR in the terminal rectum and multiple organs of ARM male rats induced by in utero exposure to DBP. DBP was administered to pregnant rats to establish the model and the incidence of ARMs in male offspring was 39.5%. On postnatal day(PND)1, the gross photograph and histopathological staining confirmed the abnormal manifestations in these organs of newborn ARMs. Decreased anogenital distance, body weight and serum testosterone level were observed in ARM male offspring. The reduced expression of Fgf10/Fgfr2 mRNA and protein was seen in terminal rectum and kidney, spleen, liver, heart in ARM male rats, whereas the reduced expression of AR was only observed in the kidney and terminal rectum. Our findings suggest the potential involvement of altered Fgf10/Fgfr2 signaling and AR in pathogenesis of local and systemic development defects in ARMs male rats induce by DBP.