Effects of Mifepristone Compound on the Receptors of Estrogen and Progesterone in Early Pregnancy Deciduas

To examine the effect of mifepristone compound (mifepristone + anor- drin) on estrogen receptor (ER) and progesterone receptor (PR) in early pregnancy decidua. Materials & Methods A Controlled study was carried out among 60 normal early pregnant volunteers (≤49d) in the department of obstetric and gynecology of Peking Union Medical Hospital. The concentrations of ER and PR were measured by radio- ligand and were compared with the control subjects after oral administration of mifepristone or mifepristone compound in different doses. Results The concentration of PR decreased while that of ER increased significantly in the decidua from all subjects administrated with mifepristone compound. We also found the concentration of EcR in Group 5 (mifepristone 30 mg + AF-53 5 mg) was the highest among 6 groups. The compound may be in favor of estrogen's action on endometrium. Conclusion The results indicate that mifepristone compound with AF- 53 has a coordi- nated function and can change the proportion of PR and ER. Hence, it can facilitate abortion. The compound dose of mifepristone 30 mg + AF-53 5mg is in favor of the endometrium recovering.

Studies on Expression of IGF-II Gene in Deciduas Derived from Medical Abortion Patients

To determine the effect of insulin like growth factor Ⅱ(IGF Ⅱ) upon the maintenance of decidua in early pregnancy and its relationship with progesterone, as well as its role in medical abortion. Materials & Methods Decidua tissue was obtained from 28 women who undergoing surgical abortion and 39 for medical abortion respectively at 5~7 weeks of gestation. The extracted total RNA was reversely transcripted and amplified by PCR with specific primers (IGF Ⅱ and β actin). The products were semi quantitated by MIAS 300 system and qualitatively analyzed by southern blotting. Results The expression of IGF Ⅱ gene in decidua from surgical abortion was significantly higher than that from medical abortion (P<0.05). The average IGF Ⅱ gene transcription values were 1.54±0.79 and 0.72±0.39 respectively. The results of southern blotting proved qualitatively that the RT PCR products were IGF Ⅱ cDNA. Conclusion IGF Ⅱ plays a role in the maintenance of decidua in early pregnancy. It may act as a mediator of progestin. It's also involved in the molecular mechanism of mifepristone.

Patterns of Ips cembrae(Herr)infestation on trap and standing trees of European larch(Larix decidua Mill.)

The large larch beetle,Ips cembrae,is a significant pest causing the death of larch.In 2020,the attack density of I.cembrae on larch trap trees and standing trees was evaluated using sample sections placed along the trunk.As a defensive measure against I.cembrae,trap trees were highly effective in both spring and summer.The attack density increased with increasing trap tree surface area/volume.Galleries were established evenly throughout the entire trunk including the thin upper portion.When the number of trap trees was low and their capacity full,a continual aggregation of adults occurred due to pheromone communication,leading to attacks on healthy standing trees in the immediate vicinity.It was found that I.cembrae attacked standing trees from the trunk base,with a continual colonization of the stem up to 70%of the tree height in a time-differentiated progression of development stages.The attack density of I.cembrae on standing trees was up to 40%lower than on the trap trees.

Effect of Mifepristone on Transforming Growth Factor -βand its Receptor Transcription in Decidua, Villi and Serum Tumor Necrosis Factor -α Level in Early Human Gestation

Objective To investigate the role of mifepristone in regulating cytokines of materno-fetal interface and serum of human early gestationMethods Thirty-five women with early pregnancy received mifepristone 50 mg orallyon study d 1 and d 2, respectively, followed by undergoing artificial abortion to getdecidua and villi on study d 3. Twenty-five women with early pregnancy withoutmifepristone administration as control also underwent artificial abortion to get de-cidua and villi. The expressions of TGF-β1 and TGF-β1 receptor mRNA in the earlydecidua and villi were assessed by using RT-PCR . The concentrations of serum TNF-α were measured by radioimmunoassay.Results The decidual expressions of TGF- β1 mRNA and TGF-β1 receptor mRNA in thetreated group were significantly lower than those of the control (P<0.05), while thevillus expressions of TGF-β1 and TGF-β1 receptor mRNA in the treated group were notsignificantly different from those of the control (P>0.05). The serum TNF-β1 levelselevated significantly after mifepristone treatment.Conclusion The antigestational effect of mifepristone might act through suppressingthe transcription of TGF-β1 and TGF-β1 receptor in the decidua and increasing theserum TNF-α level, which interfered in the materno-fetal interface Th2 bias.

In-vitro cytotoxic activity of β-Sitosterol triacontenate isolated from Capparis decidua(Forsk.) Edgew

Objective:To study the isolation and characterization of the constituent responsible for the cytotoxic activity of the ethanolic extract of stem of Capparis decidua（C.decidua）.Methods:The preliminary cytotoxic effect of isolated compound（β-Sitosterol triacontenate） was investigated by MTT assay on A549 solid tumor cells.Results:IC50 value of theβ-Sitosterol triacontenate was found to be 1μM.The cytotoxic activity increased in a dose dependent manner in case ofβ-Sitosterol triacontenate.Conclusions:The data therefore provide direct evidence for the role ofβ-Sitosterol triacontenate as a potent antimetastatic agent,which can markedly inhibit the metastatic and invasive capacity of malignant cells.

Effect of Mifepristone on the Telomerase Activity in Chorion and Decidua during Early Pregnancy

Objective To investigate telomerase activity in chorion and decidua from abortion induced by mifepristone incorporated with misoprostol at early pregnancy. Methods TRAP-SYBR Green assay was used to detect the expression of telomerase. Forty specimen were obtained from medicinal abortion （experiment group） and forty were from normal induced abortion （control group）. Results Positive expression of chorion telomerase was significantly different between the experimental group （28%, 11/40） and the control group （73%, 29/40） （P〈0.05）. While in decidua, the positive rate was 28% （11/40） in the experimental group and 20% （9/40） in the control group, there was no significant difference （P〉0.05）. Conclusion It is suggested that miferistone may significantly decrease the telomerase activity in chorion but not in decidua.

Expression of Survivin in Early Villus and Decidua and Its Implication

Summary: To investigate the expression and implication of survivin protein and mRNA in decidua and villus and the effects of mifepristone on its expression, survivin levels in decidua and villus collected from 15 normal early pregnant women and 15 early pregnant women pretreated with 150 mg mifepristone and 400 μg misoprostol were assessed by immuno-histochemical techniques and reverse transcriptional-polymerase chain reaction (RT-PCR). Our results showed that survivin proteins were stained in the cytoplasm of trophoblasts and decidual cells and in the nuclei of some of the decidual glandular epithelial cells. The expression was strongest in the trophoblasts and decidual glandular epithelial cells. The expression values in the villus and decidua were (14.56±2.44) and (10.46±2.81) respectively for normal pregnant and (8.45±2.08), (7.33±1.91) for those pretreated with mifepristone respectively (P<0.05). The transcription of survivin mRNA in villus and decidua of those pretreated with mifepristone decreased significantly compared with those in the normal pregnant women (P<0.05). It is concluded that survivin can be expressed in the decidua and villus and mifepristone inhibits its mRNA transcription and protein expression, which could possibly be one of the factors inducing decidual and villous apoptosis.

The Effects of Anordrin on Luteal Cells, DecidualCellsand TrophoblastCells in Vitro

Division of Reproductive Pharmacology, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai 200031, China *DEPT. Of Pharmacology, Shanghai Tiedao University, Medical College Shanghai 200070, China By using the morphology and the viability of cells index, the direct effects of anordrin on serum free primary cultures of rat luteal cells, human decidual cells and trophoblast cells were observed.Meanwhile, the effect of anordrin on the secretive function of rat luteal cells was also observed. The results indicated that (1) anordrin has damaging effects on rat luteal cells, human decidual cells and trophoblast cells. The LD 50 s were 14.34±0.9 μg/ml, 17.33±4.1 μg/ml and 34.87±4.9 μg/ml respectively. (2) With nonlethal dose (5 μg/ml), the activity of progesterone secretion of rat lutein cells which was stimulated by hCG and pregnenolone was not influenced by anordrin while the stimulating activity of forskolin was inhibited remarkably.The results suggest that luteolytic action is the main mechanism of the termination of early pregnancy by anordrin and the direct damaging effects of anordrin on decidua and cytotrophoblasts also play a role in the termination of early pregnancy.

Effect of Mifepristone and Anordrin Compound on Levels of Estrogen and Progesterone Receptor mRNAs in Human Decidua of Early Pregnancy

To provide the theoretical fundation for the further clinical application of mifepristone and anordrin compound. Materials & Methods Ribonuclease protection assay was used for the detection and quantitation of estrogen and progesterone receptor mRNAs in human decidua from the termination of early pregnancy.Three groups, each of which had 6～8 cases, were studied. Results Compared to the normal control group, estrogen and progesterone receptor mRNAs increased significantly (P<0.05) in the mifepristone group, whereas the changes in the group administrated mifepristone compound which contains anordrin were not obvious. Conclusions The result suggests that with the similar clinical effect, mifepristone compound has less side effect on the patients, thus being more suitable for the anti early pregnancy drug.

The Mechanism of Swainsonine Causing Early Pregnancy Abnormal Decidualization and Inducing Abortion by Changing Glycosylation Modification

Locoweed is a poisonous plant that severely harms the development of grass-land and animal husbandry throughout the world. Oxytropis and Astragalus are the Latin names for the poisonous plants commonly known as locoweed. The main toxic substance in these plants is swainsonine. It is a strong inhibitor of lysosomal α-mannosidase I and Golgi body α-mannosidase II, resulting in the accumulation of oligosaccharides and glycoproteins in lysosomes, interfering with the molecular and cellular recognition of glycans, and causing tissue-cell vacuolar degeneration. Livestock shows a series of clinical and pathological symptoms, mainly for the decline of reproductive performance. The effects in dams are abortion, weak fetus, and fetal malformations. The key step in maintaining pregnancy in dams is early endometrial decidualization, because reproductive hormones, decidual cell apoptosis, endoplasmic reticulum (ER) stress, and other immune factors are negatively affected by swainsonine. This article addresses the modifications that occur through glycan processing and glycosylation resulting in the change of post-translational modification of the protein and the activity of the glycoprotein, and then direct and indirect mechanisms of ER stress, apoptosis, reproductive hormones, immune factors, and cell cycle and other pathways. Our aim is to find new methods of prevention and treatment of swainsonine poisoning in grassland animals.

The Effect of Human Recombinant Interferon Gamma（hrIFN－γ）on hCG Secretion of Trophoblast and Protein Synthesis of Decidual Tissu

In this study the effect of human recombinant interferon gamma hrIFN-γ)on hCG secretion of human first trimester trophoblast and protein synthesis of decidual tissue was investigated in vitro.The results indicated that hrIFN-γat the doses of 250 U/ml medium and 2500 U/ml medium decreased hCG secretion of trophoblast obviously(P<0.05, P<0.01)in a dose dependent manner.The effect of hrIFN-γon protein synthesis at the doses of 10 U to 1,000 U/ml medium inhibited the 3H leucine incorporation obviously.The cpm values between control and experimental groups were significantly different(P<0. 05) in a dosedependent manner.Furthermore its inhibitory effect is in a dose-dependent manner and was neutralized by IFN-γantiserum.The IFN-a at the doses used did not find any effect on protein synthesis in decidual tissue.

Affiliation of Dihydrolipoyl Dehydrogenase Allozymes in Mycorrhizae of European Forest Trees and Characterization of the Enzyme of the Matt Bolete (<i>Xerocomus pruinatus</i>) and the Bay Bolete (<i>X. badius</i>)

Mycorrhizal roots of the deciduous trees European beech (Fagus sylvatica (L.)) and Sessile oak (Quercus petraea (MattuschkaLiebl.)) and the conifers Norway spruce (Picea abies (L.) H. Karst.) and European larch (Larix decidua (Mill.)) associated with the ectomycorrhizal fungi matt bolete (Xerocomus pruinatus (Fries 1835)) or bay bolete (X. badius (Fries 1818)) were analysed with respect to the occurrence of dihydrolipoyl dehydrogenase (EC 1.8.1.4) allozymes. In root tissues of the two deciduous trees, two gene loci could be visualized after cellulose acetate electrophoresis while three loci were expressed in root tissues of the two coniferous species. The two fungal species and further ectomycorrhizal fungi expressed exclusively one dihydrolipoyl dehydrogenase gene. In Xerocomus pruinatus and X. badius, the dihydrolipoyl dehydrogenase gene consists of 1460 bp and 1370 bp, respectively, including five introns each consisting of 52 bp. Their DNA sequences correspond to 70 to 90% to other fungal dihydrolipoyl dehydrogenase genes. One monomer of the dimeric dihydrolipoyl dehydrogenase enzyme consists of 486 (X. pruinatus) or 454 (X. badius) amino acids which sum up to a molecular mass of 55 kDa (X. pruinatus), respectively 52 kDa (X. badius). The number of positively charged amino acid residues makes 79 (X. pruinatus) and 68 (X. badius) and the number of negatively charged amino acid residues was calculated to make 46 (X. pruinatus) and 48 (X. badius);isoelectric points make 9.99 (X. pruinatus) and 9.68 (X. badius). Calculated three dimensional structures reveal a short NADH binding site being part of a larger FAD-binding site and a binding/dimerization domain.

Single-cell sequencing analysis of the maternal-fetal interface

The microenvironment at the maternal-fetal interface is optimized to facilitate the development and survival of the fetus during pregnancy.It involves a balance between cell populations and interactions of the fetal placenta with various cell types(ie,stromal cells,endothelial cells,immune cells,and fibroblasts)that are embedded in the maternal endometrium/decidua.Aberrant shifts in cell populations and deranged cell-cell interactions are closely related to pregnancy disorders.Thus,analysis of the dynamic changes in cell populations and their interactions at the maternal-fetal interface in normal and complicated pregnancies is essential to provide insights into the fundamental processes involved in the establishment and maintenance of normal pregnancy,and how these processes are dysregulated.Thus,informing novel pathways for therapeutic targets of pregnancy complications.Single-cell sequencing(SCS)is a powerful tool for transcriptome analysis at single-cell resolution.Combined with information on the developmental trajectory and function of different cell populations,SCS can provide an unparalleled opportunity for refining the spatiotemporal cell atlas to elaborate dynamic changes in cell populations and their interactions in tissues that consist of highly heterogeneous cell populations such as the maternal-fetal interface.This minireview briefly summarizes traditional methods and their limitations for analyzing maternal-fetal interface cell-cell interactions,and introduces the current applications,advantages,limitations,and prospective applications of SCS in research on maternal-fetal interactions.

Expression of Ceramide Kinase (Cerk) and Related Regulation in Early Pregnant Uterus of Mice

Objective To investigate the expression of ceramide kinase （Cerk） and related regulation in mouse uterus during early pregnancy. Methods Several mouse models, including early pregnancy, pseudopregnancy, delayed or activated implantation, artificial decidualization, and steroid hormonal treatment were performed （n=10）. Immunohistochemistry and in situ hybridization were used to detect the expression of Cerk protein and mRNA in uterus. Results Expression of Cerk mRNA and protein was strongly detected in the luminal and glandular epithelium on day 1 of pregnancy. However, Cerk mRNA and protein signals were strongly detected in the subluminal stroma surrounding the implanting blastocyst on day 5 and decidua from day 6 to day 8 whereas not in the luminal epithelium. The expression of Cerk in luminal and glandular epithelium of pseudopregnancy was similar to that of early pregnancy from day 1 to day 4 whereas on day 5 of pseudoprgenancy still with remarkable signals in the luminal epithelium. Under delayed implantation, no obvious Cerk expression was detected in the uterus. After delayed implantation was terminated by estrogen treatment and embryo implantation was initiated, both Cerk mRNA and protein were detected in the subluminal stroma surrounding the implanted blastocyst. A strong Cerk signal was detected in decidualized cells under artificial decidualization, whereas only a basal level of Cerk signal was observed in the control uterus which did not inject sesame oil. Progesterone induced a slight expression of Cerk in the luminal and glandular epithelium. Both estradiol and a combination of progesterone with estradiol strongly increased the level of Cerk signal in the luminal and glandular epithelium. Conclusion Cerk expression is under the regulation of progesterone and estrogen. The strong expression of Cerk in implantation site and decidua suggests that Cerk might play an important role during implantation and decidualization.

Natural killer cell-triggered vascular transformation:maternal care before birth?

Natural killer(NK)cells are found in lymphoid and non-lymphoid organs.In addition to important roles in immune surveillance,some NK cells contribute to angiogenesis and circulatory regulation.The uterus of early pregnancy is a non-lymphoid organ enriched in NK cells that are specifically recruited to placental attachment sites.In species with invasive hemochorial placentation,these uterine natural killer(uNK)cells,via secretion of cytokines,chemokines,mucins,enzymes and angiogenic growth factors,contribute to the physiological change of mesometrial endometrium into the unique stromal environment called decidua basalis.In humans,uNK cells have the phenotype CD56^(bright) CD16^(dim) and they appear in great abundance in the late secretory phase of the menstrual cycle and early pregnancy.Gene expression studies indicate that CD56^(bright) CD16^(dim) uterine and circulating cells are functionally distinct.In humans but not mice or other species with post-implantation decidualization,uNK cells may contribute to blastocyst implantation and are of interest as therapeutic targets in female infertility.Histological and genetic studies in mice first identified triggering of the process of gestation spiral arterial modification as a major uNK cell function,achieved via interferon(IFN)-c secretion.During spiral arterial modification,branches from the uterine artery that traverse the endometrium/decidua transiently lose their muscular coat and ability to vasoconstrict.The expression of vascular markers changes from arterial to venous as these vessels dilate and become low-resistance,high-volume channels.Full understanding of the vascular interactions of human uNK cells is difficult to obtain because endometrial time-course studies are not possible in pregnant women.Here we briefly review key information concerning uNK cell functions from studies in rodents,summarize highlights concerning human uNK cells and describe our preliminary studies on development of a humanized,pregnant mouse model for in vivo investigations of human uNK cell functions.

The integrative roles of chemokines at the maternal-feta interface in early pregnancy

Embryos express paternal antigens that are foreign to the mother, but the mother provides a special immune milieu at the fetal-maternal interface to permit rather than reject the embryo growth in the uterus until parturition by establishing precise crosstalk between the mother and the fetus. There are unanswered questions in the maintenance of pregnancy, including the poorly understood phenomenon of maternal tolerance to the allogeneic conceptus, and the remarkable biological roles of placental trophoblasts that invade the uterine wall. Chemokines are multifunctional molecules initially described as having a role in leukocyte trafficking and later found to participate in developmental processes such as differentiation and directed migration. It is increasingly evident that the gestational uterine microenvironment is characterized, at least in part, by the differential expression and secretion of chemokines that induce selective trafficking of leukocyte subsets to the maternal-fetal interface and regulate multiple events that are closely associated with normal pregnancy. Here, we review the expression and function of chemokines and their receptors at the maternal-fetal interface, with a special focus on chemokine as a key component in trophoblast invasiveness and placental angiogenesis, recruitment and instruction of immune cells so as to form a fetus-supporting milieu during pregnancy. The chemokine network is also involved in pregnancy complications.

Decidual natural killer cells and the immune microenvironment at the maternal-fetal interface

During early pregnancy,an orchestrated evolutionary maternal adaption toward tolerance of the semiallogeneic fetus is required to ensure decidualization and early embryo development.Remodeling of the immune system involves natural killer cells(NKs),macrophages,T cells and dendritic cells(DCs) altering the microenvironment in the deciduas.In particular,a unique population of NK cells with a CD56^(bright)CD16^- phenotype in the decidua has been proposed to play a key role in the maternal adaptation to pregnancy.However,there is a tendency for pregnancy immunology to reflect transplantation immunology regarding the assumption that the maternal immune system should be suppressed.This tendency is misleading.We discuss how the immune system is formed in early deciduas and the interactions between maternal NK cells and fetal growth.We propose that the maternal immune response must not be fully suppressed and is even necessary for the local response of uterine NK cells.

Role of uterine natural killer cells in angiogenesis of human decidua of the first-trimester pregnancy

Decidualization is accompanied by extensive angiogenesis, which is an essential step in the maturation of new blood vessels in mammalian pregnancy. The purpose of this study was to determine a distribution of uNK cells (CD56+ uNK or CD56bright cells) in human decidua of the first-trimester pregnancy, and investigate whether uNK cells in human decidua could express vascular endothelial growth factor (VEGF-A) and/or angiopoietin2 (Ang2). Our immunohistochemical staining results demonstrated that a great amount of uNK (CD56+) cells scattered throughout the decidual stroma and near endometrial gland and spiral vessels in human decidua. The protein expression of VEGF-A and Ang2 was detected in decidual stroma cells, capillary endothelial cells and glandular cells in tissue specimens. There was a positive correlation between microvessel density (MVD) and the number of the CD56-positive uNK cells in decidual stroma, and also between the number of the CD56-positive uNK cells and VEGF-A protein expression in the tissue. In addition, we found that uNK cells in human decidua could express VEGF-A mRNA, but not Ang2 mRNA, in isolated uNK cells in human decidua of the first-trimester gestation by combination of LCM and Nested-PCR. Our study indicated that uNK cells, through expressing VEGF-A, may play an important role in the angiogenic response at the time of human decidualization and early placenta development.

Leukocyte driven-decidual angiogenesis in early.pregnancy

Successful pregnancy and long-term, post-natal maternal and offspring cardiac, vascular and metabolic health require key maternal cardiovascular adaptations over gestation. Within the pregnant decidualizing uterus, coordinated vascular, immunological and stromal cell changes occur. Considerable attention has been given to the roles of uterine natural killer （uNK） cells in initiating decidual spiral arterial remodeling, a process normally completed by mid-gestation in mice and in humans. However, leukocyte roles in much earlier, region specific, decidual vascular remodeling are now being defined. Interest in immune cell-promoted vascular remodeling is driven by vascular aberrations that are reported in human gestational complications such as infertility, recurrent spontaneous abortion, preeclampsia （PE） and fetal growth restriction. Appropriate maternal cardiovascular responses during pregnancy protect mothers and their children from later cardiovascular disease risk elevation. One of the earliest uterine responses to pregnancy in species with hemochorial placentation is stromal cell decidualization, which creates unique niches for angiogenesis and leukocyte recruitment. In early decidua basalis, the aspect of the implantation site that will cradle the developing placenta and provide the major blood vessels to support mature placental functions, leukocytes are greatly enriched and display specialized properties. UNK cells, the most abundant leukocyte subset in early decidua basalis, have angiogenic abilities and are essential for normal early decidual angiogenesis. The regulation of uNK cells and their roles in determining maternal and progeny cardiovascular health over ore^nancv and PostPartum are discussed.

Biology and pathology of the uterine micro environ merit and its natural killer cells

Tissues are the new frontier of discoveries in immunology.Cells of the immune system are an integral part of tissue physiology and immunity.Determining how immune cells inhabit,housekeep,and defend gut,lung,brain,liver,uterus,and other organs helps revealing the intimate details of tissue physiology and may offer new therapeutic targets to treat pathologies.The uterine microenvironment modulates the development and function of innate lymphoid cells[ILC,largely represented by natural killer(NK)cells],macrophages,T cells,and dendritic cells.These immune cells,in turn,contribute to tissue homeostasis.Regulated by ovarian hormones,the human uterine mucosa(endometrium)undergoes ~400 monthly cycles of breakdown and regeneration from menarche to menopause,with its fibroblasts,glands,blood vessels,and immune cells remodeling the tissue into the transient decidua.Even more transformative changes occur upon blastocyst implantation.Before the placenta is formed,the endometrial glands feed the embryo by histiotrophic nutrition while the uterine spiral arteries are stripped of their endothelial layer and smooth muscle actin.This arterial remodeling is carried out by invading fetal trophoblast and maternal immune cells,chiefly uterine NK(uNK)cells,which also assist fetal growth.The tran sformed arteries no Ion ger resp ond to mater nal stimuli and meet the increasi ng dema nds of the growing fetus.This review focuses on how the everchanging uterine microenvironment affects uNK cells and how uNK cells regulate homeostasis of the decidua,placenta development,and fetal growth.Determining these pathways will help understand the causes of major pregnancy complications.