Objective To investigate the sensitivity of the human sperm motility assay for detecting endotoxin in culture mediumMaterials &. Methods Motile sperm were separated and exposed to different concentrations of endot...Objective To investigate the sensitivity of the human sperm motility assay for detecting endotoxin in culture mediumMaterials &. Methods Motile sperm were separated and exposed to different concentrations of endotoxin (0.5 ng/mL, 1ng/mL, 10ng/mL, 1000ng/mL, 10 000ng/ mL, and 50 000ng/mL), and sperm motility was determined after incubation. Effects of endotoxin on sperm motility in media without albumin were also examined. In addition, at the same concentrations of endotoxin (0. 5ng/mL, 1 ng/mL, and 10 ng/ mL ) , the sensitivity of the human sperm motility assay was compared to those of 1-cell and 2-cell mouse embryo bioassays.Results At levels of 0. 5ng/mL-1000ng/mL endotoxin in media with 2mg/mL albumin, sperm did not show significant change in motility during 24 h of incubation when compared with the control (P>0. 05). However, the sperm motility was significantly inhibited at endotoxin dosages of 10 000 and 50 000 ng/mL. In the absence of albumin supplementation, at endotoxin levels of 50 000ng/mL, and 1 000ng/mL, there was a marked decrease in sperm motility compared with the control after 2 h or 8 h of incubation, respectively (P<0. 01). In media containing 0. 5 ng/mL and 1 ng/ mL endotoxin, 1-cell and 2-cell mouse embryos had significantly reduced developmental rates in all developmental stages, and at the level of 10ng/mL, the development of the embryos was arrested. Conclusion The human sperm motility assay could detect high levels of endotoxin in culture medium but its sensitivity to endotoxin would be inferior to that of the 1-cell or 2-cell mouse embryo bioassay. In the absence of albumin supplementation, the sensitivity of the sperm motility assay could be improved.展开更多
To investigate the effect of clenbuterol hydrochloride on the in vitro development of both 1 cell and 2 cell mouse embryos. Methods The cultural systems of both 1 cell and 2 cell mouse embryo were used to dete...To investigate the effect of clenbuterol hydrochloride on the in vitro development of both 1 cell and 2 cell mouse embryos. Methods The cultural systems of both 1 cell and 2 cell mouse embryo were used to determine the effect of clenbuterol hydrochloride at doses of 1 ng/mL, 3 ng/mL, and 10 ng/mL on developmental rates of mouse embryos. Results When 1 cell embryos cultured with 1 ng/mL of clenbuterol hydrochloride, developmental rates from the 4 cell stage to blastocyst stage were significantly lower than those in the control group (P<0.05), but on dosages of 3 ng/mL and 10 ng/mL, the inhibiting effects on embryo development were significantly increased (P<0.01). When 2 cell embryos cultured with 1 ng/mL of clenbuterol hydrochloride, obvious differences in developmental rates were not found between the 2 cell embryo group and the control (P>0.05). However, at levels of 3 ng/mL and 10 ng/mL, significant decrease of developmental rates in 2 cell embryos was observed from the 4 cell and from the 8 cell stage, respectively (P<0.05). Embryos cultured with clenbuterol hydrochloride appeared to have more granules, fragments and degeneration than those in the control. Conclusion Clenbuterol hydrochloride has a toxic effect on the mouse embryos, and the effect is in a dose dependent. 1 cell mouse embryos cultured with clenbuterol hydrochloride could be easily inhibited at 2 cell stage, but the effect of clenbuterol hydrochloride on development of the late 2 cell embryos would be reduced.展开更多
Objective To investigate the expression and change of the β3 integrin subunit and fibronectin in normal human oviductal tissue during various phases of the menstrual cycle and tubal ectopic pregnant tissueMethods Sam...Objective To investigate the expression and change of the β3 integrin subunit and fibronectin in normal human oviductal tissue during various phases of the menstrual cycle and tubal ectopic pregnant tissueMethods Samples of normal ( n=29 ) and pregnant fallopian tube ( n=22 ) tissues were obtained from women who had normal cycle and history of normal pregnancy. Normal oviductal tissue samples were divided into 4 groups based on their menstrual cycle. Both expression and distribution of the β3 subunit and fibronectin were determined with the immunohistochemical method and the image analysis.Results The β3 subunit was expressed in the cytoplasm of ciliated cells. The expression level of the β3 subunit was higher after ovulation than that before ovulation in isthmus epithelium (P<0.001), and declined significantly after ovulation in ampullae epithelium (P<0.001). In umbrella epithelium within 4 days after ovulation, the expression level of the β3 subunit was observed at rather higher level among other phases (P <0.001). The ciliated and secretory cells of the epithelium except for where the pregnancy occurred in tubal pregnancy expressed the β3 subunit, and no significant relationship was found between the normal tubal tissue of the secretory phase and tubal ectopic pregnant tissue (P>0.05). Fibronectin was expressed in the basement membrane of human oviductal epithelium and matrix. The expression level of fibronectin was higher in the hyperplastic phase than that in the secretory one (P<0.001). And it was lower in normal tubal tissue of the secretory phase than that in tubal ectopic pregnant tissue (P<0.001).Conclusion Theβ3 integrin subunit was expressed in the ciliated cells of human oviductal epithelium, and fibronectin was expressed in the basement membrane of human oviductal epithelium and matrix. Their expression and change in oviductal tissue is based on different phases of menstrual cycle. The β3 subunit could not related to the occurrence of tubal ectopic pregnancy. Fibronectin could be the potential molecular basis for the tubal ectopic pregnancy.展开更多
Objective: To evaluate the efficiency of Percoll density gradient and swim-up methods for motile sperm isolation for in vitro fertilization and embryo transfer (IVF-ET) program. Methods: The fertilization rate, cleava...Objective: To evaluate the efficiency of Percoll density gradient and swim-up methods for motile sperm isolation for in vitro fertilization and embryo transfer (IVF-ET) program. Methods: The fertilization rate, cleavage rate, embryo developing status and pregnancy outcome of 362 IVF cycles using sperm obtained by the two methods were studied. Results: There was no significant difference in fertilization rate and cleavage rate between the Percoll and swim-up groups. Although the two groups showed no significant difference in the embryo cell number, the percentage of embryos with<20% debris was significantly higher in the Percoll group (77.6%) than in swim-up group (65.9%). The pregnancy rate and the life birth rate were also significantly higher (P<0.01) in the Percoll group (43.7% and 70.3%, respectively) than in the swim-up group (36.6 % and 60.7 %, respectively). Conclusion: The efficiency of the Percoll density gradient method is superior to the swim-up method in motile sperm separation for the IVF-ET program. [Reprod Contracep (in Chinese) 2002; 22: 292]展开更多
Objective: To evaluate the sensitivity of sperm motility assay for detecting the endotoxin effect on human sperm in vitro. Methods: Motile human sperm were separately incubated for up to 24 hours with different concen...Objective: To evaluate the sensitivity of sperm motility assay for detecting the endotoxin effect on human sperm in vitro. Methods: Motile human sperm were separately incubated for up to 24 hours with different concentrations of endotoxin (0.5, 1, 10, 1000, 10 000 and 50 000 ng/mL). Then the sperm motility was determined. The effect of endotoxin on the sperm motility in media without albumin was also determined. In addition, at the endotoxin concentrations of 0.5, 1 and 10 ng/mL, the sensitivity of the assay was compared to those of 1-cell and 2-cell mouse embryo bioassays. Results: At levels of 0.5-1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility after 24 h of incubation (P>0.05), while it was significantly inhibited at endotoxin levels of 10 000 and 50 000 ng/mL. In media without albumin, endotoxin levels of 50 000 and 1 000 ng/mL, markedly inhibited the sperm motility after 2 or 8 h of incubation (P<0.01). With media containing 0.5 and 1 ng/mL endotoxin, there was a significant reduction in the development rate at all developmental stages with 1-cell and 2-cell mouse embryo assays and at the level of 10 ng/mL, the embryo development was completely arrested. Conclusion: The sperm motility assay could detect high levels of endotoxin effect in vitro, but its sensitivity is low as compared with the 1-cell or 2-cell mouse embryo bioassay.展开更多
基金This study was supported by the Science&Technology Commission of Guangdong Province,P.R.China
文摘Objective To investigate the sensitivity of the human sperm motility assay for detecting endotoxin in culture mediumMaterials &. Methods Motile sperm were separated and exposed to different concentrations of endotoxin (0.5 ng/mL, 1ng/mL, 10ng/mL, 1000ng/mL, 10 000ng/ mL, and 50 000ng/mL), and sperm motility was determined after incubation. Effects of endotoxin on sperm motility in media without albumin were also examined. In addition, at the same concentrations of endotoxin (0. 5ng/mL, 1 ng/mL, and 10 ng/ mL ) , the sensitivity of the human sperm motility assay was compared to those of 1-cell and 2-cell mouse embryo bioassays.Results At levels of 0. 5ng/mL-1000ng/mL endotoxin in media with 2mg/mL albumin, sperm did not show significant change in motility during 24 h of incubation when compared with the control (P>0. 05). However, the sperm motility was significantly inhibited at endotoxin dosages of 10 000 and 50 000 ng/mL. In the absence of albumin supplementation, at endotoxin levels of 50 000ng/mL, and 1 000ng/mL, there was a marked decrease in sperm motility compared with the control after 2 h or 8 h of incubation, respectively (P<0. 01). In media containing 0. 5 ng/mL and 1 ng/ mL endotoxin, 1-cell and 2-cell mouse embryos had significantly reduced developmental rates in all developmental stages, and at the level of 10ng/mL, the development of the embryos was arrested. Conclusion The human sperm motility assay could detect high levels of endotoxin in culture medium but its sensitivity to endotoxin would be inferior to that of the 1-cell or 2-cell mouse embryo bioassay. In the absence of albumin supplementation, the sensitivity of the sperm motility assay could be improved.
基金the Natural Science Foundation of Guangdong ProvinceChina
文摘To investigate the effect of clenbuterol hydrochloride on the in vitro development of both 1 cell and 2 cell mouse embryos. Methods The cultural systems of both 1 cell and 2 cell mouse embryo were used to determine the effect of clenbuterol hydrochloride at doses of 1 ng/mL, 3 ng/mL, and 10 ng/mL on developmental rates of mouse embryos. Results When 1 cell embryos cultured with 1 ng/mL of clenbuterol hydrochloride, developmental rates from the 4 cell stage to blastocyst stage were significantly lower than those in the control group (P<0.05), but on dosages of 3 ng/mL and 10 ng/mL, the inhibiting effects on embryo development were significantly increased (P<0.01). When 2 cell embryos cultured with 1 ng/mL of clenbuterol hydrochloride, obvious differences in developmental rates were not found between the 2 cell embryo group and the control (P>0.05). However, at levels of 3 ng/mL and 10 ng/mL, significant decrease of developmental rates in 2 cell embryos was observed from the 4 cell and from the 8 cell stage, respectively (P<0.05). Embryos cultured with clenbuterol hydrochloride appeared to have more granules, fragments and degeneration than those in the control. Conclusion Clenbuterol hydrochloride has a toxic effect on the mouse embryos, and the effect is in a dose dependent. 1 cell mouse embryos cultured with clenbuterol hydrochloride could be easily inhibited at 2 cell stage, but the effect of clenbuterol hydrochloride on development of the late 2 cell embryos would be reduced.
基金This is a part of the project supported by Natural Science Foundation of Guangdong Province,China.
文摘Objective To investigate the expression and change of the β3 integrin subunit and fibronectin in normal human oviductal tissue during various phases of the menstrual cycle and tubal ectopic pregnant tissueMethods Samples of normal ( n=29 ) and pregnant fallopian tube ( n=22 ) tissues were obtained from women who had normal cycle and history of normal pregnancy. Normal oviductal tissue samples were divided into 4 groups based on their menstrual cycle. Both expression and distribution of the β3 subunit and fibronectin were determined with the immunohistochemical method and the image analysis.Results The β3 subunit was expressed in the cytoplasm of ciliated cells. The expression level of the β3 subunit was higher after ovulation than that before ovulation in isthmus epithelium (P<0.001), and declined significantly after ovulation in ampullae epithelium (P<0.001). In umbrella epithelium within 4 days after ovulation, the expression level of the β3 subunit was observed at rather higher level among other phases (P <0.001). The ciliated and secretory cells of the epithelium except for where the pregnancy occurred in tubal pregnancy expressed the β3 subunit, and no significant relationship was found between the normal tubal tissue of the secretory phase and tubal ectopic pregnant tissue (P>0.05). Fibronectin was expressed in the basement membrane of human oviductal epithelium and matrix. The expression level of fibronectin was higher in the hyperplastic phase than that in the secretory one (P<0.001). And it was lower in normal tubal tissue of the secretory phase than that in tubal ectopic pregnant tissue (P<0.001).Conclusion Theβ3 integrin subunit was expressed in the ciliated cells of human oviductal epithelium, and fibronectin was expressed in the basement membrane of human oviductal epithelium and matrix. Their expression and change in oviductal tissue is based on different phases of menstrual cycle. The β3 subunit could not related to the occurrence of tubal ectopic pregnancy. Fibronectin could be the potential molecular basis for the tubal ectopic pregnancy.
文摘Objective: To evaluate the efficiency of Percoll density gradient and swim-up methods for motile sperm isolation for in vitro fertilization and embryo transfer (IVF-ET) program. Methods: The fertilization rate, cleavage rate, embryo developing status and pregnancy outcome of 362 IVF cycles using sperm obtained by the two methods were studied. Results: There was no significant difference in fertilization rate and cleavage rate between the Percoll and swim-up groups. Although the two groups showed no significant difference in the embryo cell number, the percentage of embryos with<20% debris was significantly higher in the Percoll group (77.6%) than in swim-up group (65.9%). The pregnancy rate and the life birth rate were also significantly higher (P<0.01) in the Percoll group (43.7% and 70.3%, respectively) than in the swim-up group (36.6 % and 60.7 %, respectively). Conclusion: The efficiency of the Percoll density gradient method is superior to the swim-up method in motile sperm separation for the IVF-ET program. [Reprod Contracep (in Chinese) 2002; 22: 292]
文摘Objective: To evaluate the sensitivity of sperm motility assay for detecting the endotoxin effect on human sperm in vitro. Methods: Motile human sperm were separately incubated for up to 24 hours with different concentrations of endotoxin (0.5, 1, 10, 1000, 10 000 and 50 000 ng/mL). Then the sperm motility was determined. The effect of endotoxin on the sperm motility in media without albumin was also determined. In addition, at the endotoxin concentrations of 0.5, 1 and 10 ng/mL, the sensitivity of the assay was compared to those of 1-cell and 2-cell mouse embryo bioassays. Results: At levels of 0.5-1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility after 24 h of incubation (P>0.05), while it was significantly inhibited at endotoxin levels of 10 000 and 50 000 ng/mL. In media without albumin, endotoxin levels of 50 000 and 1 000 ng/mL, markedly inhibited the sperm motility after 2 or 8 h of incubation (P<0.01). With media containing 0.5 and 1 ng/mL endotoxin, there was a significant reduction in the development rate at all developmental stages with 1-cell and 2-cell mouse embryo assays and at the level of 10 ng/mL, the embryo development was completely arrested. Conclusion: The sperm motility assay could detect high levels of endotoxin effect in vitro, but its sensitivity is low as compared with the 1-cell or 2-cell mouse embryo bioassay.
