Background:Replacement gilts are typically fed ad libitum,whereas emerging evidence from human and rodent studies has revealed that time-restricted access to food has health benefits.The objective of this study was to...Background:Replacement gilts are typically fed ad libitum,whereas emerging evidence from human and rodent studies has revealed that time-restricted access to food has health benefits.The objective of this study was to investigate the effect of meal frequency on the metabolic status and ovarian follicular development in gilts.Methods:A total of 36 gilts(Landrace×Yorkshire)with an age of 150±3 d and a body weight of 77.6±3.8 kg were randomly allocated into one of three groups(n=12 in each group),and based on the group allocation,the gilts were fed at a frequency of one meal(T1),two meals(T2),or six meals per day(T6)for 14 consecutive weeks.The effects of the meal frequency on growth preference,nutrient utilization,short-chain fatty acid production by gut microbial,the post-meal dynamics in the metabolic status,reproductive hormone secretions,and ovarian follicular development in the gilts were measured.Results:The gilts in the T1 group presented a higher average daily gain(+48 g/d,P<0.05)and a higher body weight(+4.9 kg,P<0.05)than those in the T6 group.The meal frequency had no effect on the apparent digestibility of dry matter,crude protein,ether extract,ash,and gross energy,with the exception that the T1 gilts exhibited a greater NDF digestibility than the T6 gilts(P<0.05).The nitrogen balance analysis revealed that the T1 gilts presented decreased urine excretion of nitrogen(−8.17 g/d,P<0.05)and higher nitrogen retention(+9.81 g/d,P<0.05),and thus exhibited higher nitrogen utilization than the T6 gilts.The time-course dynamics of glucose,α-amino nitrogen,urea,lactate,and insulin levels in serum revealed that the T1 group exhibited higher utilization of nutrients after a meal than the T2 or T6 gilts.The T1 gilts also had a higher acetate content and SCFAs in feces than the T6 gilts(P<0.05).The age,body weight and backfat thickness of the gilts at first estrous expression were not affected by the meal frequency,but the gilts in the T1 group had higher levels of serum luteinizing hormone on the 18th day of the 3rd estrus cycle and 17β-estradiol,a larger number of growing follicles and corpora lutea,and higher mRNA expression levels of genes related to follicular development on the 19th day of the 3rd estrus cycle.Conclusions:The current findings revealed the benefits of a lower meal frequency equal feed intake on nutrient utilization and reproductive function in replacement gilts,and thus provide new insights into the nutritional strategy for replacement gilts,and the dietary pattern for other mammals,such as humans.展开更多
BACKGROUND High humidity and temperature in Taiwan have significant effects on the reproductivity of Holstein cattle,resulting in the occurrence of bovine ovarian follicular cyst(OFC).Because of economic loss from OFC...BACKGROUND High humidity and temperature in Taiwan have significant effects on the reproductivity of Holstein cattle,resulting in the occurrence of bovine ovarian follicular cyst(OFC).Because of economic loss from OFC,manual rupture and hormone injection have been advocated for the management of OFC.However,these incomplete treatments increase hormone resistance in cattle.Mesenchymal stem cells(MSCs)derived from placental stem cells(PSCs)demonstrate potential properties for the treatment of several diseases via promoting angiogenesis and immune modulation.AIM To establish the possibility of cattle placental stem cells(CPSCs)as a treatment modality for OFC of cows in Taiwan.METHODS The cows with OFC were divided into three groups:control(BC1 and BC2),hormone(H1 and H2),and CPSC(PS1 and PS2)treatment groups.In the hormone treatment group,the cows were given gonadotrophin-releasing hormone(GnRH)-prostaglandin-GnRH intramuscular injection with or without drainage of follicular fluid.In the CPSC treatment group,CPSCs were isolated from the placenta after labor.With the identification of surface antigen on stem cells,the cows were administered ovarian injection of 1×106 or 6×106 CPSCs with drainage.In all groups,OFC was scanned by ultrasound once a week for a total of seven times.The concentrations of estradiol and progesterone in serum were tested in the same period.The estrus cycle was analyzed by food intake and activity.If estrus was detected,artificial insemination was conducted.Then the cow was monitored by ultrasound for confirmation of pregnancy.RESULTS After 7 d of culture,CPSCs were successfully isolated from placental pieces.CPSCs significantly proliferated every 24 h and had high expression of MSC markers such as cluster of differentiation 44,as determined by flow cytometry.Ultrasound showed lower numbers of OFCs with drainage of follicular fluid.We achieved recovery rates of 0%,50%,50%,75%,75%and 75%in BC1,BC2,H1,H2,PS1,and PS2,respectively.Higher concentrations of progesterone were detected in the CPSC treatment groups.However,both hormone and CPSC treatment groups had no significant difference in the concentration of estradiol.The estrus rate was 0%,100%,25%,75%,75%and 75%in BC1,BC2,H1,H2,PS1,and PS2,respectively.The two fetuses were born in H2 and PS1.In brief,cows with CPSC injection achieved higher recovery,estrus,and inseminated conception rates.CONCLUSION CPSCs have efficacy in treating cows with OFC,and thus,may serve as an alternative treatment for reproductive disorders.展开更多
Objective:To determine the ovine ovarian histomorphology and follicular staging at various age periods in Awassi breed.Methods:Ovaries were collected from prenatal fetuses[gestational age(95±5)days],neonatal(day ...Objective:To determine the ovine ovarian histomorphology and follicular staging at various age periods in Awassi breed.Methods:Ovaries were collected from prenatal fetuses[gestational age(95±5)days],neonatal(day 0),and prepubertal ewe lambs(two and four months of age);each age group included six animals.Ovaries(n=12,each group)were dissected and processed for hematoxylin and eosin staining.Stained sections(n=24,each group)were imaged and utilized for histomorphology assessment,follicle measurement,and classification.Results:Prenatal ovaries were mainly enriched with primordial follicles accompanied by a lower proportion of primary follicles.In addition to primordial and primary follicles,neonatal ovaries demonstrated a proportion of centrally located multilayered and antral follicles.In comparison with neonatal ovaries,the proportion of multilayered and antral follicles was significantly higher in the ovaries of two-month-old lambs;conversely,the proportion of peripherally situated primordial follicles dramatically declined compared to that of earlier age of lamb.Although there was no statistical variation in the sizes of primordial follicles across groups,the mean diameter of the primary follicle in the prenatal ovaries was substantially smaller than in postnatal ovaries.Compared to the neonatal ovaries,the size of the multilayered and antral follicles in the prepubertal ovaries was substantially larger.Conclusions:The earliest follicular developmental stages were established prenatally whereas the advanced growth stages started in the neonatal period and greatly increased in the prepubertal period.展开更多
Objectives: Evaluation of outcome follicular aspiration compared to coating procedure for infertile women undergoing ICSI regarding frequency and severity of ovarian hyperstimulation syndrome (OHSS). Patients & Me...Objectives: Evaluation of outcome follicular aspiration compared to coating procedure for infertile women undergoing ICSI regarding frequency and severity of ovarian hyperstimulation syndrome (OHSS). Patients & Methods: Infertile PCOS women underwent controlled ovarian stimulation with luteal phase GnRH-agonist long down-regulation protocol. Women developed serum E2 level > 4000 pg/ml and/or >20 follicles of ≥10 mm in diameter were randomly divided to receive coasting strategy (Coasting group) or TVU-guided aspiration (Aspiration group). When ≥3 follicles were ≥18 mm and serum E2 level was and 10,000 IU was administered 36 h before oocyte retrieval, and ICSI was performed 72-hr thereafter and the frequency of moderate-to-severe OHSS was determined. Results: 82 women developed criteria for categorization and 21 women (25.6%) developed moderate-to-severe OHSS;5 women of aspiration and 16 of Coasting group;with significantly (P = 0.005) lower frequency in aspiration group. Twenty women developed ascites;3 women had clinically detectable, while 17 women had US detected ascites with significantly (P = 0.039) higher frequency among women that had coasting. All other manifestations of OHSS were significantly lower with aspiration procedure. Both coasting and aspiration therapy significantly reduced serum E2 and ovarian diameter on day of hCG injection compared to estimates taken at time of categorization. Conclusion: Coasting procedure prior to hCG injection could decrease incidence of OHSS and lessens its manifestation. Follicular aspiration provided more superior results and improved outcome of these women. Any of these modalities could be provided to infertile high-risk women according to the availability of experiences and patients’ selection.展开更多
为深入了解哺乳动物卵泡发育的机制,对小鼠卵巢颗粒细胞中干扰素诱导的跨膜蛋白1(interferon-induced transmembrane protein 1,Ifitm1)基因进行超表达和抑制表达分析,通过流式细胞术、荧光定量PCR、EdU法及western blot分析Ifitm1对小...为深入了解哺乳动物卵泡发育的机制,对小鼠卵巢颗粒细胞中干扰素诱导的跨膜蛋白1(interferon-induced transmembrane protein 1,Ifitm1)基因进行超表达和抑制表达分析,通过流式细胞术、荧光定量PCR、EdU法及western blot分析Ifitm1对小鼠颗粒细胞生长的影响及对小鼠排卵相关基因表达的调控作用,并用相关通路抑制剂处理颗粒细胞,探究Ifitm1影响小鼠卵泡发育及排卵的相关机制。结果显示,在小鼠卵巢颗粒细胞中成功地超表达和抑制表达了Ifitm1基因,干扰Ifitm1基因表达使细胞周期蛋白Ccnd1表达降低了63.5%,G0/G1期细胞占比也下降,使细胞阻滞在G2/M期,从而抑制颗粒细胞增殖;干扰Ifitm1基因还导致排卵标记基因Lhr、Ereg、Cyp19a1表达水平提高了1.95~6.76倍(P<0.05),并抑制PI3K/AKT信号通路上关键蛋白p-AKT(Ser473)的表达,而阻断PI3K/AKT信号通路后再抑制Ifitm1基因表达,Lhr、Ereg和Cyp19a1的mRNA水平则未出现明显改变,说明Ifitm1基因通过抑制PI3K/AKT信号通路活性影响排卵。以上结果表明,Ifitm1基因通过PI3K/AKT通路介导在小鼠颗粒细胞生长以及卵泡排卵过程中发挥重要作用。展开更多
基金This study was supported by National Key R&D Program of China(2018YFD0501005)National Natural Science Foundation of China,PR China(31772616).
文摘Background:Replacement gilts are typically fed ad libitum,whereas emerging evidence from human and rodent studies has revealed that time-restricted access to food has health benefits.The objective of this study was to investigate the effect of meal frequency on the metabolic status and ovarian follicular development in gilts.Methods:A total of 36 gilts(Landrace×Yorkshire)with an age of 150±3 d and a body weight of 77.6±3.8 kg were randomly allocated into one of three groups(n=12 in each group),and based on the group allocation,the gilts were fed at a frequency of one meal(T1),two meals(T2),or six meals per day(T6)for 14 consecutive weeks.The effects of the meal frequency on growth preference,nutrient utilization,short-chain fatty acid production by gut microbial,the post-meal dynamics in the metabolic status,reproductive hormone secretions,and ovarian follicular development in the gilts were measured.Results:The gilts in the T1 group presented a higher average daily gain(+48 g/d,P<0.05)and a higher body weight(+4.9 kg,P<0.05)than those in the T6 group.The meal frequency had no effect on the apparent digestibility of dry matter,crude protein,ether extract,ash,and gross energy,with the exception that the T1 gilts exhibited a greater NDF digestibility than the T6 gilts(P<0.05).The nitrogen balance analysis revealed that the T1 gilts presented decreased urine excretion of nitrogen(−8.17 g/d,P<0.05)and higher nitrogen retention(+9.81 g/d,P<0.05),and thus exhibited higher nitrogen utilization than the T6 gilts.The time-course dynamics of glucose,α-amino nitrogen,urea,lactate,and insulin levels in serum revealed that the T1 group exhibited higher utilization of nutrients after a meal than the T2 or T6 gilts.The T1 gilts also had a higher acetate content and SCFAs in feces than the T6 gilts(P<0.05).The age,body weight and backfat thickness of the gilts at first estrous expression were not affected by the meal frequency,but the gilts in the T1 group had higher levels of serum luteinizing hormone on the 18th day of the 3rd estrus cycle and 17β-estradiol,a larger number of growing follicles and corpora lutea,and higher mRNA expression levels of genes related to follicular development on the 19th day of the 3rd estrus cycle.Conclusions:The current findings revealed the benefits of a lower meal frequency equal feed intake on nutrient utilization and reproductive function in replacement gilts,and thus provide new insights into the nutritional strategy for replacement gilts,and the dietary pattern for other mammals,such as humans.
文摘BACKGROUND High humidity and temperature in Taiwan have significant effects on the reproductivity of Holstein cattle,resulting in the occurrence of bovine ovarian follicular cyst(OFC).Because of economic loss from OFC,manual rupture and hormone injection have been advocated for the management of OFC.However,these incomplete treatments increase hormone resistance in cattle.Mesenchymal stem cells(MSCs)derived from placental stem cells(PSCs)demonstrate potential properties for the treatment of several diseases via promoting angiogenesis and immune modulation.AIM To establish the possibility of cattle placental stem cells(CPSCs)as a treatment modality for OFC of cows in Taiwan.METHODS The cows with OFC were divided into three groups:control(BC1 and BC2),hormone(H1 and H2),and CPSC(PS1 and PS2)treatment groups.In the hormone treatment group,the cows were given gonadotrophin-releasing hormone(GnRH)-prostaglandin-GnRH intramuscular injection with or without drainage of follicular fluid.In the CPSC treatment group,CPSCs were isolated from the placenta after labor.With the identification of surface antigen on stem cells,the cows were administered ovarian injection of 1×106 or 6×106 CPSCs with drainage.In all groups,OFC was scanned by ultrasound once a week for a total of seven times.The concentrations of estradiol and progesterone in serum were tested in the same period.The estrus cycle was analyzed by food intake and activity.If estrus was detected,artificial insemination was conducted.Then the cow was monitored by ultrasound for confirmation of pregnancy.RESULTS After 7 d of culture,CPSCs were successfully isolated from placental pieces.CPSCs significantly proliferated every 24 h and had high expression of MSC markers such as cluster of differentiation 44,as determined by flow cytometry.Ultrasound showed lower numbers of OFCs with drainage of follicular fluid.We achieved recovery rates of 0%,50%,50%,75%,75%and 75%in BC1,BC2,H1,H2,PS1,and PS2,respectively.Higher concentrations of progesterone were detected in the CPSC treatment groups.However,both hormone and CPSC treatment groups had no significant difference in the concentration of estradiol.The estrus rate was 0%,100%,25%,75%,75%and 75%in BC1,BC2,H1,H2,PS1,and PS2,respectively.The two fetuses were born in H2 and PS1.In brief,cows with CPSC injection achieved higher recovery,estrus,and inseminated conception rates.CONCLUSION CPSCs have efficacy in treating cows with OFC,and thus,may serve as an alternative treatment for reproductive disorders.
文摘Objective:To determine the ovine ovarian histomorphology and follicular staging at various age periods in Awassi breed.Methods:Ovaries were collected from prenatal fetuses[gestational age(95±5)days],neonatal(day 0),and prepubertal ewe lambs(two and four months of age);each age group included six animals.Ovaries(n=12,each group)were dissected and processed for hematoxylin and eosin staining.Stained sections(n=24,each group)were imaged and utilized for histomorphology assessment,follicle measurement,and classification.Results:Prenatal ovaries were mainly enriched with primordial follicles accompanied by a lower proportion of primary follicles.In addition to primordial and primary follicles,neonatal ovaries demonstrated a proportion of centrally located multilayered and antral follicles.In comparison with neonatal ovaries,the proportion of multilayered and antral follicles was significantly higher in the ovaries of two-month-old lambs;conversely,the proportion of peripherally situated primordial follicles dramatically declined compared to that of earlier age of lamb.Although there was no statistical variation in the sizes of primordial follicles across groups,the mean diameter of the primary follicle in the prenatal ovaries was substantially smaller than in postnatal ovaries.Compared to the neonatal ovaries,the size of the multilayered and antral follicles in the prepubertal ovaries was substantially larger.Conclusions:The earliest follicular developmental stages were established prenatally whereas the advanced growth stages started in the neonatal period and greatly increased in the prepubertal period.
文摘Objectives: Evaluation of outcome follicular aspiration compared to coating procedure for infertile women undergoing ICSI regarding frequency and severity of ovarian hyperstimulation syndrome (OHSS). Patients & Methods: Infertile PCOS women underwent controlled ovarian stimulation with luteal phase GnRH-agonist long down-regulation protocol. Women developed serum E2 level > 4000 pg/ml and/or >20 follicles of ≥10 mm in diameter were randomly divided to receive coasting strategy (Coasting group) or TVU-guided aspiration (Aspiration group). When ≥3 follicles were ≥18 mm and serum E2 level was and 10,000 IU was administered 36 h before oocyte retrieval, and ICSI was performed 72-hr thereafter and the frequency of moderate-to-severe OHSS was determined. Results: 82 women developed criteria for categorization and 21 women (25.6%) developed moderate-to-severe OHSS;5 women of aspiration and 16 of Coasting group;with significantly (P = 0.005) lower frequency in aspiration group. Twenty women developed ascites;3 women had clinically detectable, while 17 women had US detected ascites with significantly (P = 0.039) higher frequency among women that had coasting. All other manifestations of OHSS were significantly lower with aspiration procedure. Both coasting and aspiration therapy significantly reduced serum E2 and ovarian diameter on day of hCG injection compared to estimates taken at time of categorization. Conclusion: Coasting procedure prior to hCG injection could decrease incidence of OHSS and lessens its manifestation. Follicular aspiration provided more superior results and improved outcome of these women. Any of these modalities could be provided to infertile high-risk women according to the availability of experiences and patients’ selection.
文摘为深入了解哺乳动物卵泡发育的机制,对小鼠卵巢颗粒细胞中干扰素诱导的跨膜蛋白1(interferon-induced transmembrane protein 1,Ifitm1)基因进行超表达和抑制表达分析,通过流式细胞术、荧光定量PCR、EdU法及western blot分析Ifitm1对小鼠颗粒细胞生长的影响及对小鼠排卵相关基因表达的调控作用,并用相关通路抑制剂处理颗粒细胞,探究Ifitm1影响小鼠卵泡发育及排卵的相关机制。结果显示,在小鼠卵巢颗粒细胞中成功地超表达和抑制表达了Ifitm1基因,干扰Ifitm1基因表达使细胞周期蛋白Ccnd1表达降低了63.5%,G0/G1期细胞占比也下降,使细胞阻滞在G2/M期,从而抑制颗粒细胞增殖;干扰Ifitm1基因还导致排卵标记基因Lhr、Ereg、Cyp19a1表达水平提高了1.95~6.76倍(P<0.05),并抑制PI3K/AKT信号通路上关键蛋白p-AKT(Ser473)的表达,而阻断PI3K/AKT信号通路后再抑制Ifitm1基因表达,Lhr、Ereg和Cyp19a1的mRNA水平则未出现明显改变,说明Ifitm1基因通过抑制PI3K/AKT信号通路活性影响排卵。以上结果表明,Ifitm1基因通过PI3K/AKT通路介导在小鼠颗粒细胞生长以及卵泡排卵过程中发挥重要作用。
