A method was developed for the simultaneous determination of four kinds of estrogens （hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol） in feed by gas chromatography-mass spectrometry （GC-MS）. After ...A method was developed for the simultaneous determination of four kinds of estrogens （hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol） in feed by gas chromatography-mass spectrometry （GC-MS）. After the sample was extracted by ethyl ether and cleaned-up on HLB phase extraction column, four kinds of estrogens were derived and quantified in gas chromatographymass spectrometry. The results showed that the linear detectable ranged from 2.5 ng· mL-1 to 250 ng· mL-1for hexoestrol and from 5 ng· mL-1 to 500 ng· mL-1 for three other estrogens with the correlation coefficients （R2） were no less than 0.990. The recoveries were in the range of 76.34%-96.33% and the relative standard deviation was no more than 22.7%. The limits of quantitation （LOQ） for all analytics were between 10 ug· kg^-1 and 20 ug· kg^-1. The method was accurate and sensitive and could meet the actual requirements for the analyses of feed samples.展开更多
Aim： To assess whether exogenous estradiol has any effect on migration of primordial germ cells （PGCs） in the chick. Methods： Fertilized eggs were treated with 17beta-estradiol （E2） （80 lag/egg） at stage X （d...Aim： To assess whether exogenous estradiol has any effect on migration of primordial germ cells （PGCs） in the chick. Methods： Fertilized eggs were treated with 17beta-estradiol （E2） （80 lag/egg） at stage X （day 0 of incubation）, stages 8-10 （incubation 30 h） and 13-15 （incubation 55 h）. Controls received vehicle （emulsion） only. Changes in PGC number were measured on different days according to developmental stages. Results： In male right gonads, but not in female left gonads, at stages 28-30 （incubation 132 h） significant decreases in the mean number of PGCs aggregating were observed compared with the controls （P 〈 0.05） while the total PGC number in the right and left gonads at each stage did not change （P 〉 0.05）. Conclusion： The present study provides evidence that E2 has significant effects on the localization of PGCs in male right, but not female left, gonads of chicken embryos at stages 28-30, compared with controls. （Asian J Andro12008 Mar; 10： 243-248）展开更多
To study the effects of 17beta-estradiol （E2 ） and progesterone （P4） levels on superovulatory response and recipients conception rate, the E2 and P4 levels of embryo donor （Borderdale） and recipient ewes （Inner...To study the effects of 17beta-estradiol （E2 ） and progesterone （P4） levels on superovulatory response and recipients conception rate, the E2 and P4 levels of embryo donor （Borderdale） and recipient ewes （Inner Mongolian sheep） were determined during multiple ovulation and em- bryo transfer （MOET）. The results showed that the E2 level increased from the day of initiation by CIDR （Controlled internal drug release devices） treatment and decreased to 3.18± 1.32 pg/ml on the day of superovulatory estrus. On the day of embryo collection, the ewes having higher levels of estrogenic peak showed better superovulatory response than the other groups having comparatively lower levels. On the day of initiation of super- ovulatory treatment, the ewes having lower levels of P4 showed better superovulatory response than those having higher levels of P4. The E2 level of the early abortion group increased from Day 0 to Day 45, while that of two pregnant recipient groups came down from Day 0 to Day 30. The ser- um P4 level of the early abortion group was significantly lower than that of the pregnant recipient groups on Day 45 （ P 〈 0.05）, but no significant difference was found between the two pregnant groups after Day 30. Therefore, the levels of E2 and P, on definite time affect superovulatory re- sponse of donors and conception rate of recipients during MOET.展开更多
BACKGROUND： Recent researches have indicated that estrogen has extensive neuroprotective effects. So some studies designed ovariectomized animal models and administrated with estrogen, so as to verify its neuroprotec...BACKGROUND： Recent researches have indicated that estrogen has extensive neuroprotective effects. So some studies designed ovariectomized animal models and administrated with estrogen, so as to verify its neuroprotective effects. OBJECTIVE： To observe the effect of 17 beta-estradiol on the content of norepinephrine （NE） and level of tumor necrosis factor （TNF） in submandibular glands of rats with sympathetic nerve injury, and analyze the dose-dependence and pathway of action. DESIGN： A randomized control animal study SETTINGS： Department of Hand Surgery, the 252 Hospital of Chinese PLA; Department of Hand Surgery Union Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： Fifty healthy female Wistar rats were randomly divided into 5 groups with 10 rats in each group： sham-operated group, ovariectomy＋6-OHDA＋saline group, ovariectomy＋6-OHDA＋17β-estradiol 50, 200 and 500 μg/kg groups. METHODS： The experiments were carried out in Tongji Medical College, Huazhong University of Science and Technology between October 2005 and March 2006. Bilateral ovaries were only exposed but not resected for the rats in the sham-operated group, but bilateral ovaries were resected in all the other groups. In the ovariectomy＋6-OHDA＋176-estradiol 50, 200 and 500 μg/kg groups, the rats were administrated with intraperitoneal injection of 6-OHDA （8 mg/kg）, and then immediately given 176-estradiol of corresponding dosages respectively, once a day for 10 days continuously. Rats in the sham-operated group and ovariectomy＋6-OHDA＋saline group were administrated with saline of the same volume. After administration, 5 rats in each group were killed to determine the NE contents in bilateral submandibular glands with high performance liquid chromatography-electrochemical detector （HPLC-ECD）, and the other 5 rats were used to determine the TNF levels in submandibular glands with enzyme-linked immunosorbant assay. MAIN OUTCOME MEASURES： The NE contents and TNF levels in submandibular glands of rats in each group were observed. RESULTS： All the 50 rats were involved in the analysis of results. （1） The NE content was obviously lower in the ovariectomy＋6-OHDA＋saline group than in the sham-operated group [（1 035±196）, （1 823±314） ng/g, P 〈 0.05], there were no significant differences between the ovariectomy＋6-OHDA＋17β-estradiol 50 μg/kg group and ovariectomy＋6-OHDA＋saline group [（1 004±253）, （1 035±196） ng/g, P 〉 0.05], but obviously higher in the ovariectomy＋6-OHDA＋17β-estradiol 200 and 500 μg/kg groups than in the ovariectomy＋6-OHDA＋saline group [（1 487±268）, （1 939±274）, （1 035±196） ng/g, P 〈 0.05]. （2） The TNF level was obviously higher in the ovariectomy＋6-OHDA＋saline group than in the sham-operated group [（3.498±0.792）, （1.893±0.533） ng/g, P 〈 0.05], there were no significant differences between the ovariectomy＋ 6-OHDA＋17β-estradiol 50 μg/kg group and ovariectomy＋6-OHDA＋saline group [（3.328 ±0.712）, （3.498±0,792） ng/g, P 〉 0.05], but obviously lower in the ovariectomy＋6-OHDA＋17β-estradiol 200 and 500 μg/kg groups than in the ovariectomy＋6-OHDA＋saline group [（2.639±0.438）, （2.016±0.619）, （3.498＋0.792） ng/g, P 〈 0.05]. CONCLUSION： Estrogen has obvious protective effect dose-dependently on 6-OHDA induced chemical sympathetic nerve terminal injury in rats, and it may play its protective role by reducing TNF level and ameliorating inflammatory reaction.展开更多
BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen o...BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen on the biological behaviors of human biliary epithelium mucosa remain unknown. The present study aimed to clarify their effects on the proliferation, apoptosis, migration and oxidative DNA damage of a human intrahepatic biliary epithelial cell (HIBEC) line in vitro. METHODS: HIBECs were co -cultured with 17 beta-estradiol (at 10(-9) mol/L, 10(-7) mol/L, and 10(-5) mol/L) and H. pylori (at MOI=0.5:1, 1:1, and 2:1) and continuously passaged until the 15th generation (approximately 45 days). Then, the following assays were performed. HIBEC proliferation was measured using the CCK-8 assay, plate clone-formation assay and by determining Ki-67 expression with immunocytochemistry; cell apoptosis and migration were investigated using Annexin-V/PI and transwell assays, respectively; and reactive oxygen species (ROS) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) production were detected by flow cytometry and immunofluorescence staining combined with confocal laser scanning microscopy, respectively. The results were the basis for evaluating the level of oxidative stress and the related DNA damage in HIBECs. RESULTS: HIBECs maintained a normal morphology and vitality when treated with 17 beta-estradiol (at 10(-9) mol/L) and H. pylori (at MOI=0.5:1 and 1:1). 17 beta-estradiol at 10(-7) mol/L and 10(-5) mol/L and H. pylori at MOI=2:1, by contrast, caused cell death. Compared with controls, HIBECs treated with 17 beta-estradiol (10(-9) mol/L) and H. pylori (MOI=1:1) had a higher up-regulation of proliferation, Ki-67 expression, clone formation, migration activity and the expression of ROS and 8-OHdG and exhibited a down-regulation of apoptosis. The above effects were further increased when 17 beta-estradiol and H. pylori were combined (P<0.05). CONCLUSIONS: H. pylori and 17 beta-estradiol, separately or in combination, promoted cell proliferation and suppressed apoptosis of HIBECs in vitro. The above phenomena might be related to oxidative stress and its subsequent DNA damage with H. pylori and 17 beta-estradiol.展开更多
基金Supported by Fund of Harbin Provincial Education Department(2014AB3BN041)
文摘A method was developed for the simultaneous determination of four kinds of estrogens （hexoestrol, diethylstilbestrol, estrone, and 17-beta-estradiol） in feed by gas chromatography-mass spectrometry （GC-MS）. After the sample was extracted by ethyl ether and cleaned-up on HLB phase extraction column, four kinds of estrogens were derived and quantified in gas chromatographymass spectrometry. The results showed that the linear detectable ranged from 2.5 ng· mL-1 to 250 ng· mL-1for hexoestrol and from 5 ng· mL-1 to 500 ng· mL-1 for three other estrogens with the correlation coefficients （R2） were no less than 0.990. The recoveries were in the range of 76.34%-96.33% and the relative standard deviation was no more than 22.7%. The limits of quantitation （LOQ） for all analytics were between 10 ug· kg^-1 and 20 ug· kg^-1. The method was accurate and sensitive and could meet the actual requirements for the analyses of feed samples.
基金Acknowledgment This work was supported by the National Natural Science Foundation of China （No. 30270648 and No. 30500270）, Zhejiang Province Scientific and Technological Project （No. 2005C22052） and Zhejiang Province Science Foundation （No. Y304194）. We thank Dr Ji- Min Zhang of the University of California for linguistic revision of the manuscript.
文摘Aim： To assess whether exogenous estradiol has any effect on migration of primordial germ cells （PGCs） in the chick. Methods： Fertilized eggs were treated with 17beta-estradiol （E2） （80 lag/egg） at stage X （day 0 of incubation）, stages 8-10 （incubation 30 h） and 13-15 （incubation 55 h）. Controls received vehicle （emulsion） only. Changes in PGC number were measured on different days according to developmental stages. Results： In male right gonads, but not in female left gonads, at stages 28-30 （incubation 132 h） significant decreases in the mean number of PGCs aggregating were observed compared with the controls （P 〈 0.05） while the total PGC number in the right and left gonads at each stage did not change （P 〉 0.05）. Conclusion： The present study provides evidence that E2 has significant effects on the localization of PGCs in male right, but not female left, gonads of chicken embryos at stages 28-30, compared with controls. （Asian J Andro12008 Mar; 10： 243-248）
基金funded by Agricultural Research Institute"948"Program of China(2000-2005)
文摘To study the effects of 17beta-estradiol （E2 ） and progesterone （P4） levels on superovulatory response and recipients conception rate, the E2 and P4 levels of embryo donor （Borderdale） and recipient ewes （Inner Mongolian sheep） were determined during multiple ovulation and em- bryo transfer （MOET）. The results showed that the E2 level increased from the day of initiation by CIDR （Controlled internal drug release devices） treatment and decreased to 3.18± 1.32 pg/ml on the day of superovulatory estrus. On the day of embryo collection, the ewes having higher levels of estrogenic peak showed better superovulatory response than the other groups having comparatively lower levels. On the day of initiation of super- ovulatory treatment, the ewes having lower levels of P4 showed better superovulatory response than those having higher levels of P4. The E2 level of the early abortion group increased from Day 0 to Day 45, while that of two pregnant recipient groups came down from Day 0 to Day 30. The ser- um P4 level of the early abortion group was significantly lower than that of the pregnant recipient groups on Day 45 （ P 〈 0.05）, but no significant difference was found between the two pregnant groups after Day 30. Therefore, the levels of E2 and P, on definite time affect superovulatory re- sponse of donors and conception rate of recipients during MOET.
文摘BACKGROUND： Recent researches have indicated that estrogen has extensive neuroprotective effects. So some studies designed ovariectomized animal models and administrated with estrogen, so as to verify its neuroprotective effects. OBJECTIVE： To observe the effect of 17 beta-estradiol on the content of norepinephrine （NE） and level of tumor necrosis factor （TNF） in submandibular glands of rats with sympathetic nerve injury, and analyze the dose-dependence and pathway of action. DESIGN： A randomized control animal study SETTINGS： Department of Hand Surgery, the 252 Hospital of Chinese PLA; Department of Hand Surgery Union Hospital affiliated to Tongji Medical College, Huazhong University of Science and Technology. MATERIALS： Fifty healthy female Wistar rats were randomly divided into 5 groups with 10 rats in each group： sham-operated group, ovariectomy＋6-OHDA＋saline group, ovariectomy＋6-OHDA＋17β-estradiol 50, 200 and 500 μg/kg groups. METHODS： The experiments were carried out in Tongji Medical College, Huazhong University of Science and Technology between October 2005 and March 2006. Bilateral ovaries were only exposed but not resected for the rats in the sham-operated group, but bilateral ovaries were resected in all the other groups. In the ovariectomy＋6-OHDA＋176-estradiol 50, 200 and 500 μg/kg groups, the rats were administrated with intraperitoneal injection of 6-OHDA （8 mg/kg）, and then immediately given 176-estradiol of corresponding dosages respectively, once a day for 10 days continuously. Rats in the sham-operated group and ovariectomy＋6-OHDA＋saline group were administrated with saline of the same volume. After administration, 5 rats in each group were killed to determine the NE contents in bilateral submandibular glands with high performance liquid chromatography-electrochemical detector （HPLC-ECD）, and the other 5 rats were used to determine the TNF levels in submandibular glands with enzyme-linked immunosorbant assay. MAIN OUTCOME MEASURES： The NE contents and TNF levels in submandibular glands of rats in each group were observed. RESULTS： All the 50 rats were involved in the analysis of results. （1） The NE content was obviously lower in the ovariectomy＋6-OHDA＋saline group than in the sham-operated group [（1 035±196）, （1 823±314） ng/g, P 〈 0.05], there were no significant differences between the ovariectomy＋6-OHDA＋17β-estradiol 50 μg/kg group and ovariectomy＋6-OHDA＋saline group [（1 004±253）, （1 035±196） ng/g, P 〉 0.05], but obviously higher in the ovariectomy＋6-OHDA＋17β-estradiol 200 and 500 μg/kg groups than in the ovariectomy＋6-OHDA＋saline group [（1 487±268）, （1 939±274）, （1 035±196） ng/g, P 〈 0.05]. （2） The TNF level was obviously higher in the ovariectomy＋6-OHDA＋saline group than in the sham-operated group [（3.498±0.792）, （1.893±0.533） ng/g, P 〈 0.05], there were no significant differences between the ovariectomy＋ 6-OHDA＋17β-estradiol 50 μg/kg group and ovariectomy＋6-OHDA＋saline group [（3.328 ±0.712）, （3.498±0,792） ng/g, P 〉 0.05], but obviously lower in the ovariectomy＋6-OHDA＋17β-estradiol 200 and 500 μg/kg groups than in the ovariectomy＋6-OHDA＋saline group [（2.639±0.438）, （2.016±0.619）, （3.498＋0.792） ng/g, P 〈 0.05]. CONCLUSION： Estrogen has obvious protective effect dose-dependently on 6-OHDA induced chemical sympathetic nerve terminal injury in rats, and it may play its protective role by reducing TNF level and ameliorating inflammatory reaction.
基金supported by grant from the National Natural Science Foundation of China(81401932)
文摘BACKGROUND: Biliary cancers are more common in females, and previous studies have suggested that Helicobacter pylori (H. pylori) exists in the biliary system. However, the effects of H. pylori infection and estrogen on the biological behaviors of human biliary epithelium mucosa remain unknown. The present study aimed to clarify their effects on the proliferation, apoptosis, migration and oxidative DNA damage of a human intrahepatic biliary epithelial cell (HIBEC) line in vitro. METHODS: HIBECs were co -cultured with 17 beta-estradiol (at 10(-9) mol/L, 10(-7) mol/L, and 10(-5) mol/L) and H. pylori (at MOI=0.5:1, 1:1, and 2:1) and continuously passaged until the 15th generation (approximately 45 days). Then, the following assays were performed. HIBEC proliferation was measured using the CCK-8 assay, plate clone-formation assay and by determining Ki-67 expression with immunocytochemistry; cell apoptosis and migration were investigated using Annexin-V/PI and transwell assays, respectively; and reactive oxygen species (ROS) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) production were detected by flow cytometry and immunofluorescence staining combined with confocal laser scanning microscopy, respectively. The results were the basis for evaluating the level of oxidative stress and the related DNA damage in HIBECs. RESULTS: HIBECs maintained a normal morphology and vitality when treated with 17 beta-estradiol (at 10(-9) mol/L) and H. pylori (at MOI=0.5:1 and 1:1). 17 beta-estradiol at 10(-7) mol/L and 10(-5) mol/L and H. pylori at MOI=2:1, by contrast, caused cell death. Compared with controls, HIBECs treated with 17 beta-estradiol (10(-9) mol/L) and H. pylori (MOI=1:1) had a higher up-regulation of proliferation, Ki-67 expression, clone formation, migration activity and the expression of ROS and 8-OHdG and exhibited a down-regulation of apoptosis. The above effects were further increased when 17 beta-estradiol and H. pylori were combined (P<0.05). CONCLUSIONS: H. pylori and 17 beta-estradiol, separately or in combination, promoted cell proliferation and suppressed apoptosis of HIBECs in vitro. The above phenomena might be related to oxidative stress and its subsequent DNA damage with H. pylori and 17 beta-estradiol.
