Background Rosemary extract(RE)has been reported to exert antioxidant property.However,the application of RE in late-phase laying hens on egg quality,intestinal barrier and microbiota,and oviductal function has not be...Background Rosemary extract(RE)has been reported to exert antioxidant property.However,the application of RE in late-phase laying hens on egg quality,intestinal barrier and microbiota,and oviductal function has not been systematically studied.This study was investigated to detect the potential effects of RE on performance,egg quality,serum parameters,intestinal heath,cecal microbiota and metabolism,and oviductal gene expressions in late-phase laying hens.A total of 21065-week-old“Jing Tint 6”laying hens were randomly allocated into five treatments with six replicates and seven birds per replicate and fed basal diet(CON)or basal diet supplemented with chlortetracycline at 50 mg/kg(CTC)or RE at 50 mg/kg(RE50),100 mg/kg(RE100),and 200 mg/kg(RE200).Results Our results showed that RE200 improved(P<0.05)Haugh unit and n-6/n-3 of egg yolk,serum superoxide dismutase(SOD)compared with CON.No significant differences were observed for Haugh unit and n-6/n-3 of egg yolk among CTC,RE50,RE100 and RE200 groups.Compared with CTC and RE50 groups,RE200 increased serum SOD activity on d 28 and 56.Compared with CON,RE supplementation decreased(P<0.05)total cholesterol(TC)level.CTC,RE100 and RE200 decreased(P<0.05)serum interleukin-6(IL-6)content compared with CON.CTC and RE200 increased jejunal m RNA expression of ZO-1 and Occludin compared with CON.The biomarkers of cecal microbiota and metabolite induced by RE 200,including Firmicutes,Eisenbergiella,Paraprevotella,Papillibacter,and butyrate,were closely associated with Haugh unit,n-6/n-3,SOD,IL-6,and TC.PICRUSt2 analysis indicated that RE altered carbohydrate and amino acid metabolism of cecal microbiota and increased butyrate synthesizing enzymes,including 3-oxoacid Co A-transferase and butyrate-acetoacetate Co A-transferase.Moreover,transcriptomic analysis revealed that RE200 improved gene expressions and functional pathways related to immunity and albumen formation in the oviductal magnum.Conclusions Dietary supplementation with 200 mg/kg RE could increase egg quality of late-phase laying hens via modulating intestinal barrier,cecal microbiota and metabolism,and oviductal function.Overall,RE could be used as a promising feed additive to improve egg quality of laying hens at late stage of production.展开更多
Background:In vitro production of bovine embryos is a well-established technology,but the in vitro culture(IVC)system still warrants improvements,especially regarding embryo quality.This study aimed to evaluate the ef...Background:In vitro production of bovine embryos is a well-established technology,but the in vitro culture(IVC)system still warrants improvements,especially regarding embryo quality.This study aimed to evaluate the effect of extracellular vesicles(EVs)isolated from oviductal(OF)and uterine fluid(UF)in sequential IVC on the development and quality of bovine embryos.Zygotes were cultured in SOF supplemented with either BSA or EVs-depleted fetal calf serum(dFCS)in the presence(BSA-EV and dFCS-EV)or absence of EVs from OF(D1 to D4)and UF(D5 to D8),mimicking in vivo conditions.EVs from oviducts(early luteal phase)and uterine horns(mid-luteal phase)from slaughtered heifers were isolated by size exclusion chromatography.Blastocyst rate was recorded on days 7-8 and their quality was assessed based on lipid contents,mitochondrial activity and total cell numbers,as well as survival rate after vitrification.Relative mRNA abundance for lipid metabolism-related transcripts and levels of phosphorylated hormonesensitive lipase(pHSL)proteins were also determined.Additionally,the expression levels of 383 miRNA in OF-and UF-EVs were assessed by qRT-PCR.Results:Blastocyst yield was lower(P<0.05)in BSA treatments compared with dFCS treatments.Survival rates after vitrification/warming were improved in dFCS-EVs(P<0.05).EVs increased(P<0.05)blastocysts total cell number in dFCS-EV and BSA-EV compared with respective controls(dFCS and BSA),while lipid content was decreased in dFCSEV(P<0.05)and mitochondrial activity did not change(P>0.05).Lipid metabolism transcripts were affected by EVs and showed interaction with type of protein source in medium(PPARGC1B,LDLR,CD36,FASN and PNPLA2,P<0.05).Levels of pHSL were lower in dFCS(P<0.05).Twenty miRNA were differentially expressed between OF-and UF-EVs and only bta-miR-148b was increased in OF-EVs(P<0.05).Conclusions:Mimicking physiological conditions using EVs from OF and UF in sequential IVC does not affect embryo development but improves blastocyst quality regarding survival rate after vitrification/warming,total cell number,lipid content,and relative changes in expression of lipid metabolism transcripts and lipase activation.Finally,EVs miRNA contents may contribute to the observed effects.展开更多
To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3...To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyle-neimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-Nl was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the β-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.展开更多
The present paper reported the preliminary results of identification of humannonserum oviduct specific proteins. The 1D-SDS-polyacrylamide gel electrophoresis (PAGE) and 2D-SDS-PAFE in conjunction with the immunoblott...The present paper reported the preliminary results of identification of humannonserum oviduct specific proteins. The 1D-SDS-polyacrylamide gel electrophoresis (PAGE) and 2D-SDS-PAFE in conjunction with the immunoblotting assay were used in the present study. The results showed that the nonserum oviduct specific proteins with MW130, 100 and 80 kD existed in human oviduct fluid or oviductal extract. In addition, the antibody against pig oviduct antigens could more strongly cross-react with human oviduct antigens, mainly recognizing 130,116 and 100 kD proteins from human oviduct. It is suggested that in human oviduct there are some specific antigens possessing some similar epitopes to those in pig oviducts. This result seems to be consistent with predominant cross reactivity existing in antigens of porcine and human zona pellucida.展开更多
The structural change of the oviduct of freshwater shrimp (Macrobrachium nipponense) during spawning was ex- amined by electron microscopy. The oviduct wall structural characteristics seem to be influenced significant...The structural change of the oviduct of freshwater shrimp (Macrobrachium nipponense) during spawning was ex- amined by electron microscopy. The oviduct wall structural characteristics seem to be influenced significantly by the spawning process. Before the parturition and ovulation, two types of epithelial cells (types I and II) are found in the epithelium. The free surfaces of type I and type II cells have very dense long microvilli. Under the type I and type II cells, are a relatively thick layer of secreting material and a layer of mostly dead cells. After ovulation, two other types of epithelial cells (types III and IV) are found in the oviduct wall epithelium. The free surface of type III cells only has short microvilli scattered on the surface. The thick layer with secreting material and the dead cell layer disappeared at this stage. In some type III cells, the leaking out of cytoplasm from broken cell membrane led to the death of these type III cells. The transformation of all four types of epithelial cells was in the order: IV→I→II→III.展开更多
The morphological basis of bilateral lower abdominal pain was studied by means of ultrastructural observation in patients with pelvic congestion syndrome after oviductal ligation. Fallopian tubes of 14 cases were coll...The morphological basis of bilateral lower abdominal pain was studied by means of ultrastructural observation in patients with pelvic congestion syndrome after oviductal ligation. Fallopian tubes of 14 cases were collected during operation. Of them, 10 patients suffered from pelvic congestion syndrome, 4 cases were normal used as control, the 8 small segments from the tubal isthmus were removed during ligation. The essential changes of the fallopian tubes in patients with this syndrome were the marked swelling of the C-type unmyelinated nerve fibers, the decrease in density of axoplasm and in number of microtubules and microfilaments. The Schwann's cells were swollen as well. Furthermore, the mitochondria revealed mild to moderate swelling, their cristae decreased and shortened. However, the changes of the endings of efferent nerve fibers were not obvious.The ultrastructural changes of C-type unmyelinated nerve fibers except the endings of efferent nerve fibers were closely related to the bilateral lower abdominal pain in patients with this syndrome.展开更多
[ Objective] To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2 （SBD-2） in epithelial cells of ovine oviduct. [ Methods] Epithelial cells of ovine oviduct were iso...[ Objective] To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2 （SBD-2） in epithelial cells of ovine oviduct. [ Methods] Epithelial cells of ovine oviduct were isolated and cultured; and then the cultured cells at secondary generation were divided into 17β-estradiol （E2, 10^-8 tool/L） group, estrogen nuclear receptor antagonist ICI182780 （10^-7 tool/L） group, PKA antagonist KT-5720 （1 μmol/L） group, PKC antagonist H- 7（50 μmol/L） group, nuclear factor kappa B antagonist PDTC（50μmol/L） group and the blank control group （ Control ）. Firstly, different antagonists were added into corresponding antagonist groups in order to interfere the epithelial ceils of ovine oviduct for 1 h. Then, 17β-estradiol （ 10^-8 mol/L） was added into each antagonist group and E2 group for cultivation for 6 h. Finally, real-time fluorescent quantitative RT-PCR was used to detect the changes of SBD-2 mRNA expression. [ Results] 10^-8 mol/L 17β-estrodiol had significantly Up-regulatory effects on the expression of SBD-2 mRNA （P 〈 0. 05 ）. Estrogen nuclear receptor antagonist ICI182780, NF-κB antagonist PDTC and PKC antagonist H-7 could all block the Up-regnlatory effects on SBD-2. But PKA antagonist KT-5720 showed no significant effects on the Up-regulation of SBD-2 mRNA expression induced by 17β-estrodiol. [ Conclusions] SBD-2 mRNA expression induced by 17β-estrodiol in epithelial cells of ovine oviduct was mediated by estrogen nuclear receptor ICI182780, NF-κB and PKC pathways. However, PKA pathway might not participate in the Up-regulation of SBD-2 mRNA expression.展开更多
BACKGROUND To investigate the clinicopathological features of endometrial clear cell carcinoma that has invaded the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor.CASE SUMMARY A case of endomet...BACKGROUND To investigate the clinicopathological features of endometrial clear cell carcinoma that has invaded the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor.CASE SUMMARY A case of endometrial clear cell carcinoma invading the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor was collected and analyzed using pathomorphology and immunohistochemistry.Endometrial clear cell carcinoma cells were distributed in a solid nest,papillary,shoe nail-like,and glandular tube-like distribution.There was infiltrative growth,and tumor cells had clear cytoplasm and obvious nuclear heteromorphism.The cancer tissue was necrotic and mitotic.The cancer tissue invaded the right oviduct.The ipsilateral oviduct also had an adenomatoid tumor.The adenomatoid tumor was arranged in microcapsules lined with flat or cubic cells that were surrounded by smooth muscle tissue.The adenomatoid tumor cells were round in shape.CONCLUSION Clear cell carcinoma of the endometrium can invade the oviduct and occur simultaneously with tubal adenomatoid tumors.Upon pathological diagnosis,one should pay close attention to distinguishing whether an endometrial clear cell carcinoma is invading the oviduct or whether it is accompanied by an adenomatoid tumor of the oviduct.Immunohistochemistry is helpful to differentiate these two disease entities.Endometrial clear cell carcinomas express Napsin-A and P16 and are negative for estrogen receptor and progesterone receptor.The presence of endometrial clear cell carcinoma does not affect the expression of CK and calretinin in adenomatoid tumors.展开更多
The present investigation has been carried out to examine the effect of human oviductaltissue co-culture system on the development of mouse embryos in vitro. Two-cell embryos collected from superovulated mouse were co...The present investigation has been carried out to examine the effect of human oviductaltissue co-culture system on the development of mouse embryos in vitro. Two-cell embryos collected from superovulated mouse were co-cultured with human oviductal tissue suspended inHam 's F10+10% Fetal Calf Serum(F10 FCS),or,in oviductal tissue conditioned medium andF10 FCS as control.The results showed that the proportion developed into blastocyst,proportion of hatchedand the velocity of embryo development were higher in both tissue co-culture and conditionedmedium as compared with F10 FCS control. Furthermore,the velocity and percentage ofembryomic development were higher in co-culture with ampullary tissue or its conditioned medium than that of isthmus.The effects of co-culture and conditioned medium on embryo development had no significant difference. All the embryos obtained from two co-culture systemscould cleave normally.This experimental observation indicated that human oviductalepithelium might secrete some factors to promote the embryonic development in vitro.展开更多
Experiments were conducted to study the effects of dietary taurine and housing density on oviduct function in laying hens. Green-shell laying hens were randomly assigned to a free range group and two caged groups, one...Experiments were conducted to study the effects of dietary taurine and housing density on oviduct function in laying hens. Green-shell laying hens were randomly assigned to a free range group and two caged groups, one with low-density and the other with high-density housing. Each group was further divided into control（C） and taurine treatment（T） groups. All hens were fed the same basic diet except that the T groups＇ diet was supplemented with 0.1% taurine. The experiment lasted 15 d. Survival rates, laying rates, daily feed consumption, and daily weight gain were recorded. Histological changes, inflammatory mediator levels, and oxidation and anti-oxidation levels were determined. The results show that dietary taurine supplementation and reduced housing density significantly attenuated pathophysiological changes in the oviduct. Nuclear factor-κB（NF-κB） DNA binding activity increased significantly in the high-density housing group compared with the two other housing groups and was reduced by taurine supplementation. Tumor necrosis factor-α（TNF-α） m RNA expression in the high-density and low-density C and T groups increased significantly. In the free range and low-density groups, dietary taurine significantly reduced the expression of TNF-α m RNA. Supplementation with taurine decreased interferon-γ（IFN-γ） m RNA expression significantly in the low-density groups. Interleukin 4（IL-4） m RNA expression was significantly higher in caged hens. IL-10 m RNA expression was higher in the high-density C group than in the free range and low-density C groups. Supplementation with taurine decreased IL-10 m RNA expression significantly in the high-density group and increased superoxide dismutase（SOD） activity in the free range hens. We conclude that taurine has important protective effects against oviduct damage. Reducing housing density also results in less oxidative stress, less inflammatory cell infiltration, and lower levels of inflammatory mediators in the oviduct. Therefore, both dietary taurine and reduced housing density can ameliorate oviduct injury, enhance oviduct health, and promote egg production in laying hens.展开更多
The interaction of oviductal epithelial cells （OECs） with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatoz...The interaction of oviductal epithelial cells （OECs） with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatozoa previously selected by density gradient in OEC and determinate some sperm characteristics that could explain the results obtained. In this study, we assessed in vitro fertilization （IVF）, tyrosine phosphorylation, phosphatidylserine translocation, nuclear DNA fragmentation, and chromatin decondensation. Three experimental sperm groups, previously selected by Percoll gradient, were established according to the origin of the sperm used for IVF： （i） W30 group： spermatozoa were incubated with oocytes in the absence of OEC; （ii） NB group： after sperm incubation in OEC, the unbound spermatozoa were incubated with oocytes, in the absence of OEC; and （iii） B group： after sperm incubation with OEC, the bound spermatozoa were incubated with oocytes in the OEC plates. The results showed that sperm from the NB group led to a lower IVF yield, accompanied by low penetration rates （NB： 19.6%, B： 94.9%, and W30： 62.9%; P 〈 0.001） and problems of nuclear decondensation. Moreover, higher levels of tyrosine phosphorylation were observed in the NB group compared with the W30 and B groups （NB： 58.7%, B： 2.5%, and W30： 4.5%; P 〈 0.01）. A similar trend was observed in phosphatidylserine translocation （NB： 93.7%, B. 5.7%, and W30： 44.2%; P 〈 0.01）. These results demonstrate that the OEC exerts a rigorous degree of sperm selection, even within an already highly selected population of spermatozoa, and can capture the best functional spermatozoa for fertilization.展开更多
Objective:To examine the structure of the oviduct of the shrimp Penaeus monodon.Methods:The oviducts of P.monodon with three different major groups of ovarian development(Group(Gr.) 1:Stages I & V;Gr.2:Stages II &...Objective:To examine the structure of the oviduct of the shrimp Penaeus monodon.Methods:The oviducts of P.monodon with three different major groups of ovarian development(Group(Gr.) 1:Stages I & V;Gr.2:Stages II & III;and Gr.3:Stage IV) were examined by light,transmission electron,and scanning electron microscopies,respectively.Results:The epithelium of the oviduct in Gr.1 was composed of tall simple columnar cells with their basal nuclei located on the basement membrane and its thick collagen fibers.In Gr.2,the oviduct seemed to produce some substances and their epithelial cells became transitional with centrally located nuclei and formed some vacuoles.Obviously,the epithelial cells in Gr.3(at Stage IV) were disorganized,disrupted,and shed accumulated spherical secretory substances including some cellular contents into the lumen.Conclusions:The structural changes of the P.monodon oviduct were related to ovarian maturation stages(Grs.1-3).Prior to spawning,only the oviduct epithelium at ovary Stage IV produced and secreted a number of spherical secretion substances into the lumen.These substances may act as the oviductal lubricants to facilitate the spawning process.展开更多
Objective: To find an effective treatment of oviduct pregnancy. Methods: One hundred and fifteen patients of oviduct pregnancy were treated by injecting trichosanthin in cervlx. Results: One week after injection, 95.5...Objective: To find an effective treatment of oviduct pregnancy. Methods: One hundred and fifteen patients of oviduct pregnancy were treated by injecting trichosanthin in cervlx. Results: One week after injection, 95.5 % of the treated patients had their HCG turned to negative. Symptoms such as painful abdominal distention, and signs such as pelvic hydrops and vaginal bleeding, were also relieved obviously in most of the cases. Conclusion: Trichosanthin injection may be an effective treatment for oviduct pregnancy, but further improvement in its preparation, and method of medication and dosage are necessary.展开更多
Background Spermatozoa interact with oviduct secretions before fertilization in vivo but the molecular players of this dialog and underlying dynamics remain largely unknown.Our objectives were to identify an exhaustiv...Background Spermatozoa interact with oviduct secretions before fertilization in vivo but the molecular players of this dialog and underlying dynamics remain largely unknown.Our objectives were to identify an exhaustive list of sperm-interacting proteins(SIPs)in the bovine oviduct fluid and to evaluate the impact of the oviduct anatomical region(isthmus vs.ampulla)and time relative to ovulation(pre-ovulatory vs.post-ovulatory)on SIPs number and abundance.Methods Pools of oviduct fluid(OF)from the pre-ovulatory ampulla,pre-ovulatory isthmus,post-ovulatory ampulla,and post-ovulatory isthmus in the side of ovulation were collected from the slaughterhouse.Frozen-thawed bull sperm were incubated with OF or phosphate-buffered saline(control)for 60 min at 38.5℃.After protein extraction and digestion,sperm and OF samples were analyzed by nanoLC-MS/MS and label-free protein quantification.Results A quantitative comparison between proteins identified in sperm and OF samples(2333 and 2471 proteins,respectively)allowed for the identification of 245 SIPs.The highest number(187)were found in the pre-ovulatory isthmus,i.e.,time and place of the sperm reservoir.In total,41 SIPs(17%)were differentially abundant between stages in a given region or between regions at a given stage and 76 SIPs(31%)were identified in only one region×stage condition.Functional analysis of SIPs predicted roles in cell response to stress,regulation of cell motility,fertilization,and early embryo development.Conclusion This study provides a comprehensive list of SIPs in the bovine oviduct and evidences dynamic spatiotemporal changes in sperm-oviduct interactions around ovulation time.Moreover,these data provide protein candidates to improve sperm conservation and in vitro fertilization media.展开更多
基金supported by the China Postdoctoral Science Foundation(2022M723370)the Strategic Priority Research Program of the Chinese Academy of Sciences(XDA23080603)。
文摘Background Rosemary extract(RE)has been reported to exert antioxidant property.However,the application of RE in late-phase laying hens on egg quality,intestinal barrier and microbiota,and oviductal function has not been systematically studied.This study was investigated to detect the potential effects of RE on performance,egg quality,serum parameters,intestinal heath,cecal microbiota and metabolism,and oviductal gene expressions in late-phase laying hens.A total of 21065-week-old“Jing Tint 6”laying hens were randomly allocated into five treatments with six replicates and seven birds per replicate and fed basal diet(CON)or basal diet supplemented with chlortetracycline at 50 mg/kg(CTC)or RE at 50 mg/kg(RE50),100 mg/kg(RE100),and 200 mg/kg(RE200).Results Our results showed that RE200 improved(P<0.05)Haugh unit and n-6/n-3 of egg yolk,serum superoxide dismutase(SOD)compared with CON.No significant differences were observed for Haugh unit and n-6/n-3 of egg yolk among CTC,RE50,RE100 and RE200 groups.Compared with CTC and RE50 groups,RE200 increased serum SOD activity on d 28 and 56.Compared with CON,RE supplementation decreased(P<0.05)total cholesterol(TC)level.CTC,RE100 and RE200 decreased(P<0.05)serum interleukin-6(IL-6)content compared with CON.CTC and RE200 increased jejunal m RNA expression of ZO-1 and Occludin compared with CON.The biomarkers of cecal microbiota and metabolite induced by RE 200,including Firmicutes,Eisenbergiella,Paraprevotella,Papillibacter,and butyrate,were closely associated with Haugh unit,n-6/n-3,SOD,IL-6,and TC.PICRUSt2 analysis indicated that RE altered carbohydrate and amino acid metabolism of cecal microbiota and increased butyrate synthesizing enzymes,including 3-oxoacid Co A-transferase and butyrate-acetoacetate Co A-transferase.Moreover,transcriptomic analysis revealed that RE200 improved gene expressions and functional pathways related to immunity and albumen formation in the oviductal magnum.Conclusions Dietary supplementation with 200 mg/kg RE could increase egg quality of late-phase laying hens via modulating intestinal barrier,cecal microbiota and metabolism,and oviductal function.Overall,RE could be used as a promising feed additive to improve egg quality of laying hens at late stage of production.
基金supported by the Spanish Ministry of Science and Innovation (PID2019-111641RB-I00 to DR, and RTI2018-093548-B-I00 to AGA)Sáo Paulo Research Foundation,Brazil (FAPESP+4 种基金#2017/20339-3 and CNPqBrazil 304276/2018-9 to CLVL#2019/04981-2 to RM2014/22887-0 and 2015/21829-9 to JCDS)Minciencias-Colombia Postdoctoral Fellowship (848-2019) to KC-BSecretaria de Educación Superior,Ciencia y Tecnología e Innovación (SENESCYT-Ecuador) to YNC.
文摘Background:In vitro production of bovine embryos is a well-established technology,but the in vitro culture(IVC)system still warrants improvements,especially regarding embryo quality.This study aimed to evaluate the effect of extracellular vesicles(EVs)isolated from oviductal(OF)and uterine fluid(UF)in sequential IVC on the development and quality of bovine embryos.Zygotes were cultured in SOF supplemented with either BSA or EVs-depleted fetal calf serum(dFCS)in the presence(BSA-EV and dFCS-EV)or absence of EVs from OF(D1 to D4)and UF(D5 to D8),mimicking in vivo conditions.EVs from oviducts(early luteal phase)and uterine horns(mid-luteal phase)from slaughtered heifers were isolated by size exclusion chromatography.Blastocyst rate was recorded on days 7-8 and their quality was assessed based on lipid contents,mitochondrial activity and total cell numbers,as well as survival rate after vitrification.Relative mRNA abundance for lipid metabolism-related transcripts and levels of phosphorylated hormonesensitive lipase(pHSL)proteins were also determined.Additionally,the expression levels of 383 miRNA in OF-and UF-EVs were assessed by qRT-PCR.Results:Blastocyst yield was lower(P<0.05)in BSA treatments compared with dFCS treatments.Survival rates after vitrification/warming were improved in dFCS-EVs(P<0.05).EVs increased(P<0.05)blastocysts total cell number in dFCS-EV and BSA-EV compared with respective controls(dFCS and BSA),while lipid content was decreased in dFCSEV(P<0.05)and mitochondrial activity did not change(P>0.05).Lipid metabolism transcripts were affected by EVs and showed interaction with type of protein source in medium(PPARGC1B,LDLR,CD36,FASN and PNPLA2,P<0.05).Levels of pHSL were lower in dFCS(P<0.05).Twenty miRNA were differentially expressed between OF-and UF-EVs and only bta-miR-148b was increased in OF-EVs(P<0.05).Conclusions:Mimicking physiological conditions using EVs from OF and UF in sequential IVC does not affect embryo development but improves blastocyst quality regarding survival rate after vitrification/warming,total cell number,lipid content,and relative changes in expression of lipid metabolism transcripts and lipase activation.Finally,EVs miRNA contents may contribute to the observed effects.
文摘To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyle-neimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-Nl was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the β-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.
文摘The present paper reported the preliminary results of identification of humannonserum oviduct specific proteins. The 1D-SDS-polyacrylamide gel electrophoresis (PAGE) and 2D-SDS-PAFE in conjunction with the immunoblotting assay were used in the present study. The results showed that the nonserum oviduct specific proteins with MW130, 100 and 80 kD existed in human oviduct fluid or oviductal extract. In addition, the antibody against pig oviduct antigens could more strongly cross-react with human oviduct antigens, mainly recognizing 130,116 and 100 kD proteins from human oviduct. It is suggested that in human oviduct there are some specific antigens possessing some similar epitopes to those in pig oviducts. This result seems to be consistent with predominant cross reactivity existing in antigens of porcine and human zona pellucida.
基金Project (No. Grant 961102205) supported by the Science and Tech-nology Bureau of Zhejiang Province, China
文摘The structural change of the oviduct of freshwater shrimp (Macrobrachium nipponense) during spawning was ex- amined by electron microscopy. The oviduct wall structural characteristics seem to be influenced significantly by the spawning process. Before the parturition and ovulation, two types of epithelial cells (types I and II) are found in the epithelium. The free surfaces of type I and type II cells have very dense long microvilli. Under the type I and type II cells, are a relatively thick layer of secreting material and a layer of mostly dead cells. After ovulation, two other types of epithelial cells (types III and IV) are found in the oviduct wall epithelium. The free surface of type III cells only has short microvilli scattered on the surface. The thick layer with secreting material and the dead cell layer disappeared at this stage. In some type III cells, the leaking out of cytoplasm from broken cell membrane led to the death of these type III cells. The transformation of all four types of epithelial cells was in the order: IV→I→II→III.
文摘The morphological basis of bilateral lower abdominal pain was studied by means of ultrastructural observation in patients with pelvic congestion syndrome after oviductal ligation. Fallopian tubes of 14 cases were collected during operation. Of them, 10 patients suffered from pelvic congestion syndrome, 4 cases were normal used as control, the 8 small segments from the tubal isthmus were removed during ligation. The essential changes of the fallopian tubes in patients with this syndrome were the marked swelling of the C-type unmyelinated nerve fibers, the decrease in density of axoplasm and in number of microtubules and microfilaments. The Schwann's cells were swollen as well. Furthermore, the mitochondria revealed mild to moderate swelling, their cristae decreased and shortened. However, the changes of the endings of efferent nerve fibers were not obvious.The ultrastructural changes of C-type unmyelinated nerve fibers except the endings of efferent nerve fibers were closely related to the bilateral lower abdominal pain in patients with this syndrome.
基金Supported by the National Natural Science Foundation of China(31060328)the Natural Science Foundation of Inner Mongolia(No.2014BS0801)+1 种基金the Doctor's Start-up Fund in Inner Mongolia Medical University(NY2011BQ003)the Youth Entrepreneurship Foundation of Inner Mongolia Medical University(NY2010QN003)
文摘[ Objective] To investigate the mechanisms involved in the Up-regulatory effects of 17β-estrodiol on β-defensin-2 （SBD-2） in epithelial cells of ovine oviduct. [ Methods] Epithelial cells of ovine oviduct were isolated and cultured; and then the cultured cells at secondary generation were divided into 17β-estradiol （E2, 10^-8 tool/L） group, estrogen nuclear receptor antagonist ICI182780 （10^-7 tool/L） group, PKA antagonist KT-5720 （1 μmol/L） group, PKC antagonist H- 7（50 μmol/L） group, nuclear factor kappa B antagonist PDTC（50μmol/L） group and the blank control group （ Control ）. Firstly, different antagonists were added into corresponding antagonist groups in order to interfere the epithelial ceils of ovine oviduct for 1 h. Then, 17β-estradiol （ 10^-8 mol/L） was added into each antagonist group and E2 group for cultivation for 6 h. Finally, real-time fluorescent quantitative RT-PCR was used to detect the changes of SBD-2 mRNA expression. [ Results] 10^-8 mol/L 17β-estrodiol had significantly Up-regulatory effects on the expression of SBD-2 mRNA （P 〈 0. 05 ）. Estrogen nuclear receptor antagonist ICI182780, NF-κB antagonist PDTC and PKC antagonist H-7 could all block the Up-regnlatory effects on SBD-2. But PKA antagonist KT-5720 showed no significant effects on the Up-regulation of SBD-2 mRNA expression induced by 17β-estrodiol. [ Conclusions] SBD-2 mRNA expression induced by 17β-estrodiol in epithelial cells of ovine oviduct was mediated by estrogen nuclear receptor ICI182780, NF-κB and PKC pathways. However, PKA pathway might not participate in the Up-regulation of SBD-2 mRNA expression.
文摘BACKGROUND To investigate the clinicopathological features of endometrial clear cell carcinoma that has invaded the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor.CASE SUMMARY A case of endometrial clear cell carcinoma invading the right oviduct with a cooccurring ipsilateral oviduct adenomatoid tumor was collected and analyzed using pathomorphology and immunohistochemistry.Endometrial clear cell carcinoma cells were distributed in a solid nest,papillary,shoe nail-like,and glandular tube-like distribution.There was infiltrative growth,and tumor cells had clear cytoplasm and obvious nuclear heteromorphism.The cancer tissue was necrotic and mitotic.The cancer tissue invaded the right oviduct.The ipsilateral oviduct also had an adenomatoid tumor.The adenomatoid tumor was arranged in microcapsules lined with flat or cubic cells that were surrounded by smooth muscle tissue.The adenomatoid tumor cells were round in shape.CONCLUSION Clear cell carcinoma of the endometrium can invade the oviduct and occur simultaneously with tubal adenomatoid tumors.Upon pathological diagnosis,one should pay close attention to distinguishing whether an endometrial clear cell carcinoma is invading the oviduct or whether it is accompanied by an adenomatoid tumor of the oviduct.Immunohistochemistry is helpful to differentiate these two disease entities.Endometrial clear cell carcinomas express Napsin-A and P16 and are negative for estrogen receptor and progesterone receptor.The presence of endometrial clear cell carcinoma does not affect the expression of CK and calretinin in adenomatoid tumors.
文摘The present investigation has been carried out to examine the effect of human oviductaltissue co-culture system on the development of mouse embryos in vitro. Two-cell embryos collected from superovulated mouse were co-cultured with human oviductal tissue suspended inHam 's F10+10% Fetal Calf Serum(F10 FCS),or,in oviductal tissue conditioned medium andF10 FCS as control.The results showed that the proportion developed into blastocyst,proportion of hatchedand the velocity of embryo development were higher in both tissue co-culture and conditionedmedium as compared with F10 FCS control. Furthermore,the velocity and percentage ofembryomic development were higher in co-culture with ampullary tissue or its conditioned medium than that of isthmus.The effects of co-culture and conditioned medium on embryo development had no significant difference. All the embryos obtained from two co-culture systemscould cleave normally.This experimental observation indicated that human oviductalepithelium might secrete some factors to promote the embryonic development in vitro.
基金Project supported by the National Natural Science Foundation of China(Nos.31101783 and 31472164)the Special Fund for Independent Innovation of Agricultural Science and Technology in Jiangsu Province of China(No.CX(13)3070)
文摘Experiments were conducted to study the effects of dietary taurine and housing density on oviduct function in laying hens. Green-shell laying hens were randomly assigned to a free range group and two caged groups, one with low-density and the other with high-density housing. Each group was further divided into control（C） and taurine treatment（T） groups. All hens were fed the same basic diet except that the T groups＇ diet was supplemented with 0.1% taurine. The experiment lasted 15 d. Survival rates, laying rates, daily feed consumption, and daily weight gain were recorded. Histological changes, inflammatory mediator levels, and oxidation and anti-oxidation levels were determined. The results show that dietary taurine supplementation and reduced housing density significantly attenuated pathophysiological changes in the oviduct. Nuclear factor-κB（NF-κB） DNA binding activity increased significantly in the high-density housing group compared with the two other housing groups and was reduced by taurine supplementation. Tumor necrosis factor-α（TNF-α） m RNA expression in the high-density and low-density C and T groups increased significantly. In the free range and low-density groups, dietary taurine significantly reduced the expression of TNF-α m RNA. Supplementation with taurine decreased interferon-γ（IFN-γ） m RNA expression significantly in the low-density groups. Interleukin 4（IL-4） m RNA expression was significantly higher in caged hens. IL-10 m RNA expression was higher in the high-density C group than in the free range and low-density C groups. Supplementation with taurine decreased IL-10 m RNA expression significantly in the high-density group and increased superoxide dismutase（SOD） activity in the free range hens. We conclude that taurine has important protective effects against oviduct damage. Reducing housing density also results in less oxidative stress, less inflammatory cell infiltration, and lower levels of inflammatory mediators in the oviduct. Therefore, both dietary taurine and reduced housing density can ameliorate oviduct injury, enhance oviduct health, and promote egg production in laying hens.
文摘The interaction of oviductal epithelial cells （OECs） with the spermatozoa has beneficial effects on the sperm functions. The aim of this study is to evaluate the in vitro fertilizing capacity of incubating spermatozoa previously selected by density gradient in OEC and determinate some sperm characteristics that could explain the results obtained. In this study, we assessed in vitro fertilization （IVF）, tyrosine phosphorylation, phosphatidylserine translocation, nuclear DNA fragmentation, and chromatin decondensation. Three experimental sperm groups, previously selected by Percoll gradient, were established according to the origin of the sperm used for IVF： （i） W30 group： spermatozoa were incubated with oocytes in the absence of OEC; （ii） NB group： after sperm incubation in OEC, the unbound spermatozoa were incubated with oocytes, in the absence of OEC; and （iii） B group： after sperm incubation with OEC, the bound spermatozoa were incubated with oocytes in the OEC plates. The results showed that sperm from the NB group led to a lower IVF yield, accompanied by low penetration rates （NB： 19.6%, B： 94.9%, and W30： 62.9%; P 〈 0.001） and problems of nuclear decondensation. Moreover, higher levels of tyrosine phosphorylation were observed in the NB group compared with the W30 and B groups （NB： 58.7%, B： 2.5%, and W30： 4.5%; P 〈 0.01）. A similar trend was observed in phosphatidylserine translocation （NB： 93.7%, B. 5.7%, and W30： 44.2%; P 〈 0.01）. These results demonstrate that the OEC exerts a rigorous degree of sperm selection, even within an already highly selected population of spermatozoa, and can capture the best functional spermatozoa for fertilization.
文摘Objective:To examine the structure of the oviduct of the shrimp Penaeus monodon.Methods:The oviducts of P.monodon with three different major groups of ovarian development(Group(Gr.) 1:Stages I & V;Gr.2:Stages II & III;and Gr.3:Stage IV) were examined by light,transmission electron,and scanning electron microscopies,respectively.Results:The epithelium of the oviduct in Gr.1 was composed of tall simple columnar cells with their basal nuclei located on the basement membrane and its thick collagen fibers.In Gr.2,the oviduct seemed to produce some substances and their epithelial cells became transitional with centrally located nuclei and formed some vacuoles.Obviously,the epithelial cells in Gr.3(at Stage IV) were disorganized,disrupted,and shed accumulated spherical secretory substances including some cellular contents into the lumen.Conclusions:The structural changes of the P.monodon oviduct were related to ovarian maturation stages(Grs.1-3).Prior to spawning,only the oviduct epithelium at ovary Stage IV produced and secreted a number of spherical secretion substances into the lumen.These substances may act as the oviductal lubricants to facilitate the spawning process.
文摘Objective: To find an effective treatment of oviduct pregnancy. Methods: One hundred and fifteen patients of oviduct pregnancy were treated by injecting trichosanthin in cervlx. Results: One week after injection, 95.5 % of the treated patients had their HCG turned to negative. Symptoms such as painful abdominal distention, and signs such as pelvic hydrops and vaginal bleeding, were also relieved obviously in most of the cases. Conclusion: Trichosanthin injection may be an effective treatment for oviduct pregnancy, but further improvement in its preparation, and method of medication and dosage are necessary.
基金funded by INRAE and Agence Nationale de la Recherche under the grant number ANR-18-CE92-0049supported by grants from Biogenouest+1 种基金Infrastructures en Biologie Santéet Agronomie (IBiSA)Conseil Régional de Bretagne awarded to Protim proteomics core facility。
文摘Background Spermatozoa interact with oviduct secretions before fertilization in vivo but the molecular players of this dialog and underlying dynamics remain largely unknown.Our objectives were to identify an exhaustive list of sperm-interacting proteins(SIPs)in the bovine oviduct fluid and to evaluate the impact of the oviduct anatomical region(isthmus vs.ampulla)and time relative to ovulation(pre-ovulatory vs.post-ovulatory)on SIPs number and abundance.Methods Pools of oviduct fluid(OF)from the pre-ovulatory ampulla,pre-ovulatory isthmus,post-ovulatory ampulla,and post-ovulatory isthmus in the side of ovulation were collected from the slaughterhouse.Frozen-thawed bull sperm were incubated with OF or phosphate-buffered saline(control)for 60 min at 38.5℃.After protein extraction and digestion,sperm and OF samples were analyzed by nanoLC-MS/MS and label-free protein quantification.Results A quantitative comparison between proteins identified in sperm and OF samples(2333 and 2471 proteins,respectively)allowed for the identification of 245 SIPs.The highest number(187)were found in the pre-ovulatory isthmus,i.e.,time and place of the sperm reservoir.In total,41 SIPs(17%)were differentially abundant between stages in a given region or between regions at a given stage and 76 SIPs(31%)were identified in only one region×stage condition.Functional analysis of SIPs predicted roles in cell response to stress,regulation of cell motility,fertilization,and early embryo development.Conclusion This study provides a comprehensive list of SIPs in the bovine oviduct and evidences dynamic spatiotemporal changes in sperm-oviduct interactions around ovulation time.Moreover,these data provide protein candidates to improve sperm conservation and in vitro fertilization media.
