[ Objective] To study the growth-development of Boer goats from embryo transplant. [ Method] With Boer goats from embryo transplant as the tested animals and natural breeding Boer goats born in the same period as CK, ...[ Objective] To study the growth-development of Boer goats from embryo transplant. [ Method] With Boer goats from embryo transplant as the tested animals and natural breeding Boer goats born in the same period as CK, the weight of Boer goats at birth and at the age of 1,3 and 6 months as well as the body size at the age of 3 and 6 months were investigated in this study, while their health conditions were also observed and recorded. [ Result] Under the normal feeding conditions, Boer goats from embryo transplant developed well at early stage, and their meat-using body type were also obvious. The average birth weight of bucks was （4.25±0.95） kg, and that of does was （3.74±0.10） kg. The average weight of bucks at the age of 6 months was （31.90 ±0.74） kg and that of was （25.90 ±0.67） kg. The daily gains of bucks and ewes from birth to 6 months of age were 153.64 and 123.11 g respectively, while their body length, body height and chest circumference increased obviously, and there was no significant difference in body weight or body size between them and natural breeding goats. [ Condusioa] The study lays a foundation for further studies on the breedinq and utilization of Boer noats from embryo transplant at early staae.展开更多
A series of experiments were conducted to study the major procedures in nuclear transplantation such as oocyte enucleation and activation, electrofusion and developnent of the nuclear transplant embryos in the mouse, ...A series of experiments were conducted to study the major procedures in nuclear transplantation such as oocyte enucleation and activation, electrofusion and developnent of the nuclear transplant embryos in the mouse, rabbits and sheep. The important results are as follows:11. In the mouse, only 35% of the oocytes collected 15～16 h after hCG had a notable first polar body (FPb) and those without FPb were enucleated by removing cytoplasm from the PVS-wider side and the enucleation rate was similar to that in the oocytes with FPb, and the enucleation rate of removing 1/3 cytoplasm was remarkably higher than that of removing 1/4 cytoplasm. 2. Among the three fusion media tested, mannitol and sucrose solutions produced better results than M2 in electrofusion of mouse 2-cell embryos. Under favorable pulse conditions, the osmotic pressure of fusion medium had no motable effect on electrofusion, but as the conditions became so unfavorable that some embryos began to lyse, the fusion rates in hypertonie mannitol solution were significantly higher than those in isotonic or hypotonic solutions. A wide range of pulse strengths (0.31～2.04 by/ cm) and durations(10～1280 μs) were used and 100% of fusion were obtained in many cases. Optimal pulse durations were plotted for field strengths to obtain high fusion rates (96%～ 100%) in mouse2-cell embryos. 3. With one pulse of 0.45 by / cm, satisfactory results of mouse oocyte activation were obtained only when the duration increased to 160 μs or longer. The activation rate increased as the oocytes got older. Some of the oocytes ar. rested at metaphase Ⅲ after electrical stimulation and their proportion to the number of oocytes not activated increased with egg age. 4. 10% and 31% of the nuclear transplant embryos developed to morula or blastocyst stage in sheep and rabbits, respectively, with Chinese-made hormones and chemicals.展开更多
BACKGROUND： Total saponins of Panax ginseng （TSPG） exhibits neuroprotection against Parkinson＇s disease in the substantia nigra. OBJECTIVE： To investigate the effects of TSPG on human embryonic neural stem cells ...BACKGROUND： Total saponins of Panax ginseng （TSPG） exhibits neuroprotection against Parkinson＇s disease in the substantia nigra. OBJECTIVE： To investigate the effects of TSPG on human embryonic neural stem cells （NSCs） proliferation and differentiation into dopaminergic neurons using in vitro studies, and to observe NSC differentiation in a mouse model of Parkinson＇s disease, as well as behavioral changes before and after transplantation. DESIGN, TIME AND SETTING： In vitro neural cell biology trial and in vivo randomized, controlled animal trial were performed at the Institute of Basic Medical Sciences, Chongqing Medical University between September 2004 and December 2007. MATERIALS： TSPG （purity 〉 95%） was isolated, extracted, and identified by Chongqing Academy of Chinese Materia Medica. Recombinant human basic fibroblast growth factor （bFGF） and recombinant human epidermal growth factor （EGF） were purchased from PeproTech, USA. A total of 25 C57/BL6J mice, aged 18-20 weeks were included. Twenty were used to establish a Parkinson＇s disease model with i.p. injection of MPTP （1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine） and TSPG alone or combined with interleukin-1 （IL-1）-treated NSCs prior to transplantation into the corpus striatum. The remaining five mice were pretreated for 3 days with TSPG prior to MPTP injection, serving as the TSPG prevention group. METHODS： Primary NSCs were isolated, cultured and purified from embryonic cerebral cortex. Immunocytochemistry was employed to detect specific antigen expression in the NSCs. In vitro experiment： （1） to induce proliferation, NSCs were treated with TSPG, EGF＋bFGF, or TSPG＋EGF＋bFGF, respectively; （2） to induce dopaminergic neuronal differentiation, NSCs were treated with TSPG, IL-1, or TSPG＋IL-1, respectively. MAIN OUTCOME MEASURES： In vitro experiment： the effects of TSPG on NSCs proliferation were evaluated with flow cytometry and MTT assay. Tyrosine hydroxylase expression was determined by immunocytochemistry assay to observe effects of TSPG on dopaminergic neuronal differentiation. In vivo experiment： differentiation of grafted NSCs in the mouse brain was determined by immunohistochemical staining. Behavioral changes were evaluated by spontaneous activity frequency, memory function, and score of paralysis agitans. RESULTS： （1） NSCs were cultured and passaged for more than three passages. Immunocytochemistry revealed positive nestin staining, as well as neurofilament protein and glial fibrillary acidic protein. （2） TSPG significantly increased NSC proliferation, in particular when combined with EGF and bFGF, which was twice as effective as FGF or bFGF alone. TSPG also induced dopaminergic differentiation in NSCs, in particular when TSPG was added together with IL-1, resulting in an effect five times greater than that of IL-1 alone. （3） At day 30 following transplantation, most NSCs in the TSPG prevention group differentiated into dopaminergic neurons, and the scores of paralysis agitans, spontaneous activity, and memory function were significantly increased compared with TSPG alone or TSPG＋IL-1 groups （P 〈 0.05）. CONCLUSION： TSPG stimulated NSC proliferation, in particular when combined with FGF and bFGF. TSPG significantly induced dopaminergic neuronal differentiation of NSCs, and the effect was greater when combined with IL-1. In addition, TSPG greatly improved behavior in the Parkinson＇s disease mouse model following NSC transplantation. Following NSC transplantation, TSPG pretreatment exhibited superior efficacy over either TSPG alone or TSPG in combination with IL-1, in terms of behavioral improvements in the Parkinson＇s disease mouse model.展开更多
Objective:Some patients fail to obtain an embryo for transplantation during previousin vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)cycles,and require multiple reproductive treatments.This study ai...Objective:Some patients fail to obtain an embryo for transplantation during previousin vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)cycles,and require multiple reproductive treatments.This study aimed to evaluate whether changing the control ovarian stimulation(COS)protocol during the subsequent stimulation cycle could improve laboratory and clinical outcomes in these patients.Methods:Patients without a transplantable embryo(TE)in the previous IVF/ICSI cycles were recruited during their second cycles.They were classified into two groups according to their first cycle protocol:Group A,patients treated with a gonadotropinreleasing hormone agonist(GnRH-a),and Group B,patients treated with a gonadotropin-releasing hormone antagonist(GnRH-ant).The study group included patients whose stimulation protocols were changed,whereas the control group consisted of patients who used the same stimulation protocol in the second cycle.We then compared the numbers of oocytes collected(OC)and TE,the incidence of non-TE,the pregnancy rate(PR),and the live birth rate(LBR).Results:In Group A,the numbers of OC and TE were significantly lower(6.0±4.7vs.9.4±6.4,2.3±2.2vs.4.5±3.8,P<0.05)in the study group compared with those in the control group.In Group B,the numbers of OC and TE were higher(7.0±5.5vs.4.0±4.3,3.5±3.4vs.1.8±2.1,P<0.05)in the study group.There was a significant increase in the incidence of non-TE(adjusted odds ratio(AOR)=2.12,95%CI:1.04–4.69)of the study group in Group A but not in Group B.No significant differences in the PR or LBR were found between the study and control groups in either Group A or B.Conclusion:Changing the COS protocol from GnRH-ant to GnRH-a or continuing the GnRH-a protocol can improve laboratory outcomes in patients with no TE in the previous IVF/ICSI cycle.展开更多
基金supported by Talent Research Foundation of Henan University of Technology (09001095)Scientific and Technological Project of Henan Province " Research and Demonstration of Supporting Production Technology Systemfor High-quality Funiu White Goats" (072102130004)
文摘[ Objective] To study the growth-development of Boer goats from embryo transplant. [ Method] With Boer goats from embryo transplant as the tested animals and natural breeding Boer goats born in the same period as CK, the weight of Boer goats at birth and at the age of 1,3 and 6 months as well as the body size at the age of 3 and 6 months were investigated in this study, while their health conditions were also observed and recorded. [ Result] Under the normal feeding conditions, Boer goats from embryo transplant developed well at early stage, and their meat-using body type were also obvious. The average birth weight of bucks was （4.25±0.95） kg, and that of does was （3.74±0.10） kg. The average weight of bucks at the age of 6 months was （31.90 ±0.74） kg and that of was （25.90 ±0.67） kg. The daily gains of bucks and ewes from birth to 6 months of age were 153.64 and 123.11 g respectively, while their body length, body height and chest circumference increased obviously, and there was no significant difference in body weight or body size between them and natural breeding goats. [ Condusioa] The study lays a foundation for further studies on the breedinq and utilization of Boer noats from embryo transplant at early staae.
文摘A series of experiments were conducted to study the major procedures in nuclear transplantation such as oocyte enucleation and activation, electrofusion and developnent of the nuclear transplant embryos in the mouse, rabbits and sheep. The important results are as follows:11. In the mouse, only 35% of the oocytes collected 15～16 h after hCG had a notable first polar body (FPb) and those without FPb were enucleated by removing cytoplasm from the PVS-wider side and the enucleation rate was similar to that in the oocytes with FPb, and the enucleation rate of removing 1/3 cytoplasm was remarkably higher than that of removing 1/4 cytoplasm. 2. Among the three fusion media tested, mannitol and sucrose solutions produced better results than M2 in electrofusion of mouse 2-cell embryos. Under favorable pulse conditions, the osmotic pressure of fusion medium had no motable effect on electrofusion, but as the conditions became so unfavorable that some embryos began to lyse, the fusion rates in hypertonie mannitol solution were significantly higher than those in isotonic or hypotonic solutions. A wide range of pulse strengths (0.31～2.04 by/ cm) and durations(10～1280 μs) were used and 100% of fusion were obtained in many cases. Optimal pulse durations were plotted for field strengths to obtain high fusion rates (96%～ 100%) in mouse2-cell embryos. 3. With one pulse of 0.45 by / cm, satisfactory results of mouse oocyte activation were obtained only when the duration increased to 160 μs or longer. The activation rate increased as the oocytes got older. Some of the oocytes ar. rested at metaphase Ⅲ after electrical stimulation and their proportion to the number of oocytes not activated increased with egg age. 4. 10% and 31% of the nuclear transplant embryos developed to morula or blastocyst stage in sheep and rabbits, respectively, with Chinese-made hormones and chemicals.
文摘BACKGROUND： Total saponins of Panax ginseng （TSPG） exhibits neuroprotection against Parkinson＇s disease in the substantia nigra. OBJECTIVE： To investigate the effects of TSPG on human embryonic neural stem cells （NSCs） proliferation and differentiation into dopaminergic neurons using in vitro studies, and to observe NSC differentiation in a mouse model of Parkinson＇s disease, as well as behavioral changes before and after transplantation. DESIGN, TIME AND SETTING： In vitro neural cell biology trial and in vivo randomized, controlled animal trial were performed at the Institute of Basic Medical Sciences, Chongqing Medical University between September 2004 and December 2007. MATERIALS： TSPG （purity 〉 95%） was isolated, extracted, and identified by Chongqing Academy of Chinese Materia Medica. Recombinant human basic fibroblast growth factor （bFGF） and recombinant human epidermal growth factor （EGF） were purchased from PeproTech, USA. A total of 25 C57/BL6J mice, aged 18-20 weeks were included. Twenty were used to establish a Parkinson＇s disease model with i.p. injection of MPTP （1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine） and TSPG alone or combined with interleukin-1 （IL-1）-treated NSCs prior to transplantation into the corpus striatum. The remaining five mice were pretreated for 3 days with TSPG prior to MPTP injection, serving as the TSPG prevention group. METHODS： Primary NSCs were isolated, cultured and purified from embryonic cerebral cortex. Immunocytochemistry was employed to detect specific antigen expression in the NSCs. In vitro experiment： （1） to induce proliferation, NSCs were treated with TSPG, EGF＋bFGF, or TSPG＋EGF＋bFGF, respectively; （2） to induce dopaminergic neuronal differentiation, NSCs were treated with TSPG, IL-1, or TSPG＋IL-1, respectively. MAIN OUTCOME MEASURES： In vitro experiment： the effects of TSPG on NSCs proliferation were evaluated with flow cytometry and MTT assay. Tyrosine hydroxylase expression was determined by immunocytochemistry assay to observe effects of TSPG on dopaminergic neuronal differentiation. In vivo experiment： differentiation of grafted NSCs in the mouse brain was determined by immunohistochemical staining. Behavioral changes were evaluated by spontaneous activity frequency, memory function, and score of paralysis agitans. RESULTS： （1） NSCs were cultured and passaged for more than three passages. Immunocytochemistry revealed positive nestin staining, as well as neurofilament protein and glial fibrillary acidic protein. （2） TSPG significantly increased NSC proliferation, in particular when combined with EGF and bFGF, which was twice as effective as FGF or bFGF alone. TSPG also induced dopaminergic differentiation in NSCs, in particular when TSPG was added together with IL-1, resulting in an effect five times greater than that of IL-1 alone. （3） At day 30 following transplantation, most NSCs in the TSPG prevention group differentiated into dopaminergic neurons, and the scores of paralysis agitans, spontaneous activity, and memory function were significantly increased compared with TSPG alone or TSPG＋IL-1 groups （P 〈 0.05）. CONCLUSION： TSPG stimulated NSC proliferation, in particular when combined with FGF and bFGF. TSPG significantly induced dopaminergic neuronal differentiation of NSCs, and the effect was greater when combined with IL-1. In addition, TSPG greatly improved behavior in the Parkinson＇s disease mouse model following NSC transplantation. Following NSC transplantation, TSPG pretreatment exhibited superior efficacy over either TSPG alone or TSPG in combination with IL-1, in terms of behavioral improvements in the Parkinson＇s disease mouse model.
基金grants to W.H.M.from the National Natural Science Foundation of China(81860160)M.J.L.from Guangxi Key Research and Development Program(No.AB20238002)+2 种基金Y.H.Y.from the National Natural Science Foundation of China(81871172)the Natural Science Foundation of Guangxi Zhuang Autonomous Region(2019GXNSFFA245013,2018GXNSFDA050017)the Guangxi Medical University Training Program for Distinguished Young Scholars and the Special Fund of the Female Fertility Preservation Innovation Team of the First Affiliated Hospital of Guangxi Medical University。
文摘Objective:Some patients fail to obtain an embryo for transplantation during previousin vitro fertilization(IVF)or intracytoplasmic sperm injection(ICSI)cycles,and require multiple reproductive treatments.This study aimed to evaluate whether changing the control ovarian stimulation(COS)protocol during the subsequent stimulation cycle could improve laboratory and clinical outcomes in these patients.Methods:Patients without a transplantable embryo(TE)in the previous IVF/ICSI cycles were recruited during their second cycles.They were classified into two groups according to their first cycle protocol:Group A,patients treated with a gonadotropinreleasing hormone agonist(GnRH-a),and Group B,patients treated with a gonadotropin-releasing hormone antagonist(GnRH-ant).The study group included patients whose stimulation protocols were changed,whereas the control group consisted of patients who used the same stimulation protocol in the second cycle.We then compared the numbers of oocytes collected(OC)and TE,the incidence of non-TE,the pregnancy rate(PR),and the live birth rate(LBR).Results:In Group A,the numbers of OC and TE were significantly lower(6.0±4.7vs.9.4±6.4,2.3±2.2vs.4.5±3.8,P<0.05)in the study group compared with those in the control group.In Group B,the numbers of OC and TE were higher(7.0±5.5vs.4.0±4.3,3.5±3.4vs.1.8±2.1,P<0.05)in the study group.There was a significant increase in the incidence of non-TE(adjusted odds ratio(AOR)=2.12,95%CI:1.04–4.69)of the study group in Group A but not in Group B.No significant differences in the PR or LBR were found between the study and control groups in either Group A or B.Conclusion:Changing the COS protocol from GnRH-ant to GnRH-a or continuing the GnRH-a protocol can improve laboratory outcomes in patients with no TE in the previous IVF/ICSI cycle.
