In the present study, the effect of blockage of the costimulatory signal CD86 at time of implantation on the expressions of TGF-β1, MMP-9, TIMP-3 and PAI-1 proteins at the maternal-fetal interface and the outcome of ...In the present study, the effect of blockage of the costimulatory signal CD86 at time of implantation on the expressions of TGF-β1, MMP-9, TIMP-3 and PAI-1 proteins at the maternal-fetal interface and the outcome of pregnancy in murine abortion-prone model was investigated, in which the CBA/J x DBA/2 matings were used as the abortion-prone model and the CBA/J×BALB/c matings used as the normal pregnant model. The study was performed in following three groups: 2 groups of the abortion-prone model, which were experimental group and control experimental group, and 1 group of normal pregnant model, and each group had 10 pregnant CBA/J mice exclusively. Female pregnant CBA/J mice in the experimental group received an intraperitoneal (i. p.) injection of 100μg of antimouse CD86 mAb in 200μl of PBS at day 4.5 of gestation, and the irrelevant-isotope matched rat IgG2b was administrated in the control experimental group with the same dosage and at same time. For the normal pregnant group, no treatment was given. The pregnant CBA/J mice were killed on day 13.5 of gestation. Then, the embryo resorption rate was calculated and the expressions of TGF-β1, MMP-9, TIMP3 and PAI-1 were detected by using immunohistochemical methods. It was demonstrated that the embryo resorption rate in the experimental group was significantly reduced in comparison with that in the control experimental group (x2=7.441,P = 0.006), but there was no significant difference with that in normal pregnant group (x2=0.016, P = 0.898) . The expressions of TGF-β1 and PAI-1 in the experimental group were significantly increased in comparison with that in the control experimental group (P=0.010,P=0.003, respectively), with no significant difference from that in the normal pregnant group (P = 0.500) . However, the expression of MMP-9 in the experimental group was significantly reduced in comparison with that in the control experimental group (P = 0.012) with no significant difference from that in the normal pregnant group (P = 0.500) . The expression of TIMP-3 in the experimental group showed no significant difference both with the control experimental group (P = 0. 328) and the normal pregnant group (P = 0.500) . It is concluded that the blockage of the costimulatory molecule CD86 at early stage of gestation can render TGF-β1, MMP-9, TIMP-3 and PAI-1 proteins to express their immuno-tolerant effects through their characteristic pathways and induce the reduction of the embryo resorption rate in the natural abortion-prone model of mice to the level of normal pregnancy.展开更多
Objective: To explore the mechanism of ShouTaiWan's intervention in the treatment of Threatened Abortion based on Network Pharmacology. Methods: The effective active ingredients and targets in ShouTaiWan were obta...Objective: To explore the mechanism of ShouTaiWan's intervention in the treatment of Threatened Abortion based on Network Pharmacology. Methods: The effective active ingredients and targets in ShouTaiWan were obtained by searching in TCMSP database, BATMAN-TCM database and Uniprot database. The target of threatened abortion was obtained by searching for 'threatened abortion' in GeneCards. And the co-target gene of ShouTaiWan and threaten abortion can be obtained through the use of gene mapping in Venn diagram. Then an interaction network "disease-drug-target" can be constructed through a software called Cytoscape. Upload the Co-targeted gene to the STRING database to construct the protein interaction network map (PPI) , which would be screened for the key genes by using R language. Finally, used the R language to analyze gene ontology (GO) enrichment and kyoto encyclopedia of genes and genome (KEGG) pathway enrichment. Result: It is predicted that 21 effective active ingredients for the treatment of threatened abortion and 89 common effective targets are expected. Quercetin, kaempferol, beta-sitosterol, and isorhamnetin were the most effective components. As for the targets, prostaglandin endoperoxidase receptor (PTGS2), progesterone receptor (PGR), prostaglandin endoperoxidase receptor 1 (PTGS1), and Peroxisome proliferator activates receptor gamma (PPARG) were the most effective ones. The core targets in PPI network are Interleukin-6 (IL-6), protein kinase (AKT1), vascular endothelial growth factor A (VEGFA), etc. GO enrichment analysis shows that the activity of cytokines, regulatory functions, nuclear protein receptors and transcription of genes would be affected. KEGG pathway enrichment analysis illustrates that advanced glycation end products-receptor for AGE (AGE-RAGE) pathways are of the highest significance, followed by the fluid shear stress and atherosclerosis pathways. Conclusion: ShouTaiWan can achieve anti-inflammatory, increase vascular permeability,improve endometrial receptivity and other aspects of regulation to treat threatened abortion and embryo implantation through regulating cytokine.展开更多
Objective:To study the association of environmental adverse factors with early embryo loss,and explore the possible risk factors in daily life. Methods:A questionnaire was administered to 93 new cases of embryo loss(c...Objective:To study the association of environmental adverse factors with early embryo loss,and explore the possible risk factors in daily life. Methods:A questionnaire was administered to 93 new cases of embryo loss(case group) collected in four general hospitals in Tianjin from April 2007 to April 2008 and 93 matched cases of induced abortion(control group) in normal pregnant women who sought the abortion by other reasons.The questionnaire covered information on parental exposure to various environmental factors during and before pregnancy,and the information on daily life.Data were analyzed by single-factor analysis,multiple linear regression and logistic regression analysis.Possible risk factors were identified and odds ratio calculated. Results:Cooking frequently during pregnancy,more daily traffic hours,and decoration history in early pregnancy and paternal exposure to toxic matters three months before pregnancy were associated with early embryo loss, while maternal education was a protective factor. Conclusion:Women exposed to the harmful substances from traffic emissions,cooking and decoration could be at an increased risk of early embryo loss.展开更多
A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductin...A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductins. N-terminal peptide (NTP) (384 residues) and C-terminal peptide (CTP) (73 residues) of deduced protein precursor has about 80% and 50% identity with that of other mammals respectively. Fusion proteins GST-NTP 368(1R-368N)and GST-CTP73 (369F-441A) were expressed and purified. NH2-terminal of CTP sequencing reveals that the purified protein is consistent with the deduced one. In order to study the function of NTP and CTP the mouse anti-NTP and rabbit anti-CTP antisera were prepared. Tissue-specific (skeleton muscle, oviduct, uterus, ovary, liver, heart and brain) analysis indicated that rabbit oviductin was only found in oviduct. The conditioned medium derived from the rabbit oviduct mucosa epithelial cells has a function of overcoming the early embryonic development block of Kunming mous e cultured in vitro. Anti-CTP antiserum could totally inhibit the early embryo development at 2-cell stage cultured in the conditioned culture medium, but anti-NTP antiserum couldn’t. There was a positive relationship between the ratio of early embryos at development block and the dosage of anti-CTP antiserum added in the conditioned culture medium. These results suggest that oviductin has a function not only on fertilization, but also on the release of early embryonic development block, and the later function domain of rabbit oviductin may be situate in its C-terminal.展开更多
文摘In the present study, the effect of blockage of the costimulatory signal CD86 at time of implantation on the expressions of TGF-β1, MMP-9, TIMP-3 and PAI-1 proteins at the maternal-fetal interface and the outcome of pregnancy in murine abortion-prone model was investigated, in which the CBA/J x DBA/2 matings were used as the abortion-prone model and the CBA/J×BALB/c matings used as the normal pregnant model. The study was performed in following three groups: 2 groups of the abortion-prone model, which were experimental group and control experimental group, and 1 group of normal pregnant model, and each group had 10 pregnant CBA/J mice exclusively. Female pregnant CBA/J mice in the experimental group received an intraperitoneal (i. p.) injection of 100μg of antimouse CD86 mAb in 200μl of PBS at day 4.5 of gestation, and the irrelevant-isotope matched rat IgG2b was administrated in the control experimental group with the same dosage and at same time. For the normal pregnant group, no treatment was given. The pregnant CBA/J mice were killed on day 13.5 of gestation. Then, the embryo resorption rate was calculated and the expressions of TGF-β1, MMP-9, TIMP3 and PAI-1 were detected by using immunohistochemical methods. It was demonstrated that the embryo resorption rate in the experimental group was significantly reduced in comparison with that in the control experimental group (x2=7.441,P = 0.006), but there was no significant difference with that in normal pregnant group (x2=0.016, P = 0.898) . The expressions of TGF-β1 and PAI-1 in the experimental group were significantly increased in comparison with that in the control experimental group (P=0.010,P=0.003, respectively), with no significant difference from that in the normal pregnant group (P = 0.500) . However, the expression of MMP-9 in the experimental group was significantly reduced in comparison with that in the control experimental group (P = 0.012) with no significant difference from that in the normal pregnant group (P = 0.500) . The expression of TIMP-3 in the experimental group showed no significant difference both with the control experimental group (P = 0. 328) and the normal pregnant group (P = 0.500) . It is concluded that the blockage of the costimulatory molecule CD86 at early stage of gestation can render TGF-β1, MMP-9, TIMP-3 and PAI-1 proteins to express their immuno-tolerant effects through their characteristic pathways and induce the reduction of the embryo resorption rate in the natural abortion-prone model of mice to the level of normal pregnancy.
基金Special research project of Chinese medicine industry(No.201507001-02)National administration of traditional Chinese medicine(No.1199ws02)
文摘Objective: To explore the mechanism of ShouTaiWan's intervention in the treatment of Threatened Abortion based on Network Pharmacology. Methods: The effective active ingredients and targets in ShouTaiWan were obtained by searching in TCMSP database, BATMAN-TCM database and Uniprot database. The target of threatened abortion was obtained by searching for 'threatened abortion' in GeneCards. And the co-target gene of ShouTaiWan and threaten abortion can be obtained through the use of gene mapping in Venn diagram. Then an interaction network "disease-drug-target" can be constructed through a software called Cytoscape. Upload the Co-targeted gene to the STRING database to construct the protein interaction network map (PPI) , which would be screened for the key genes by using R language. Finally, used the R language to analyze gene ontology (GO) enrichment and kyoto encyclopedia of genes and genome (KEGG) pathway enrichment. Result: It is predicted that 21 effective active ingredients for the treatment of threatened abortion and 89 common effective targets are expected. Quercetin, kaempferol, beta-sitosterol, and isorhamnetin were the most effective components. As for the targets, prostaglandin endoperoxidase receptor (PTGS2), progesterone receptor (PGR), prostaglandin endoperoxidase receptor 1 (PTGS1), and Peroxisome proliferator activates receptor gamma (PPARG) were the most effective ones. The core targets in PPI network are Interleukin-6 (IL-6), protein kinase (AKT1), vascular endothelial growth factor A (VEGFA), etc. GO enrichment analysis shows that the activity of cytokines, regulatory functions, nuclear protein receptors and transcription of genes would be affected. KEGG pathway enrichment analysis illustrates that advanced glycation end products-receptor for AGE (AGE-RAGE) pathways are of the highest significance, followed by the fluid shear stress and atherosclerosis pathways. Conclusion: ShouTaiWan can achieve anti-inflammatory, increase vascular permeability,improve endometrial receptivity and other aspects of regulation to treat threatened abortion and embryo implantation through regulating cytokine.
文摘Objective:To study the association of environmental adverse factors with early embryo loss,and explore the possible risk factors in daily life. Methods:A questionnaire was administered to 93 new cases of embryo loss(case group) collected in four general hospitals in Tianjin from April 2007 to April 2008 and 93 matched cases of induced abortion(control group) in normal pregnant women who sought the abortion by other reasons.The questionnaire covered information on parental exposure to various environmental factors during and before pregnancy,and the information on daily life.Data were analyzed by single-factor analysis,multiple linear regression and logistic regression analysis.Possible risk factors were identified and odds ratio calculated. Results:Cooking frequently during pregnancy,more daily traffic hours,and decoration history in early pregnancy and paternal exposure to toxic matters three months before pregnancy were associated with early embryo loss, while maternal education was a protective factor. Conclusion:Women exposed to the harmful substances from traffic emissions,cooking and decoration could be at an increased risk of early embryo loss.
基金Supported by National Natural Science Foundation of China (39730460)National "973" Project (G1999055902)National Labora-
文摘A full-length rabbit oviductin cDNA(1909bp) was cloned. It consists of a 5’-UTR of 52bp, an open reading frame (ORF) of 1374bp and a 3’-UTR of 483bp and has more than 80% homology with that of other mammal oviductins. N-terminal peptide (NTP) (384 residues) and C-terminal peptide (CTP) (73 residues) of deduced protein precursor has about 80% and 50% identity with that of other mammals respectively. Fusion proteins GST-NTP 368(1R-368N)and GST-CTP73 (369F-441A) were expressed and purified. NH2-terminal of CTP sequencing reveals that the purified protein is consistent with the deduced one. In order to study the function of NTP and CTP the mouse anti-NTP and rabbit anti-CTP antisera were prepared. Tissue-specific (skeleton muscle, oviduct, uterus, ovary, liver, heart and brain) analysis indicated that rabbit oviductin was only found in oviduct. The conditioned medium derived from the rabbit oviduct mucosa epithelial cells has a function of overcoming the early embryonic development block of Kunming mous e cultured in vitro. Anti-CTP antiserum could totally inhibit the early embryo development at 2-cell stage cultured in the conditioned culture medium, but anti-NTP antiserum couldn’t. There was a positive relationship between the ratio of early embryos at development block and the dosage of anti-CTP antiserum added in the conditioned culture medium. These results suggest that oviductin has a function not only on fertilization, but also on the release of early embryonic development block, and the later function domain of rabbit oviductin may be situate in its C-terminal.
