OBJECTIVETo explore the differences of metabolic footprint in the conditioned culture medium of placental explants between early-onset and late-onset severe preeclampsia.

METHODSIn 13 cases of early-onset severe preeclampsia and 14 cases of late-onset severe preeclampsia, the placentas were sampled at the surface of the maternal placenta. High performance liquid chromatography-mass spectrometry (HPLC-MS) was used to determine the differences in the metabolites in the conditioned culture medium of the placental villous explants cultured in 6% atmospheric O(2) for 96 h. Standard samples were used to establish the tryptophan and kynurenine chromatography library by HPLC-MS to analyze the concentration of tryptophan and kynurenine in the conditioned culture medium.

RESULTSThirty-six metabolites showed statistically significant differences between early-onset and late-onset severe preeclampsia (P<0.05). The concentration of kynurenine was significantly higher in early-onset severe preeclampsia than in late-onset severe preeclampsia (P<0.05).

CONCLUSIONEarly-onset and late-onset severe preeclampsia may have different pathogeneses. By detecting the concentration of metabolites, metabolomic strategies provide a new means for predicting the onset time of severe preeclampsia.

Chorionic Villi ; metabolism ; Culture Media, Conditioned ; chemistry ; Female ; Humans ; In Vitro Techniques ; Kynurenine ; metabolism ; Ornithine ; metabolism ; Placenta ; metabolism ; Pre-Eclampsia ; metabolism ; Pregnancy ; Tryptophan ; metabolism

OBJECTIVETo investigate the roles of thromboxane A(2) (TXA(2)) and prostaglandin I(2) (PGI(2)) in development of oligohydramnios.

METHODSThe concentration of TXB(2) and 6-keto-PGF1 in umbilical cord blood collected from 30 normal parturients (control) and 30 parturients with oligohydramnios was detected by radioimmunoassay to calculate the TXA(2)/PGI(2) ratio. Immunohistochemistry was performed to detect the contents of TXA(2)R in vascular endothelial cell in the placental villi.

RESULTSCompared with the control group, the concentration of umbilical cord blood TXB(2) in oligohydramnios group was significantly increased (P<0.01), but the elevation of 6-keto-PGF(2) concentration was not statistically significant (P>0.05). The oligohydramnios group showed significantly higher positivity rates of TXB2 and 6-keto-PGF1 in than the control group (P<0.01), and the positivity rate of TXA(2)R in the vascular endothelial cells in the placental villi was also significantly higher in the oligohydramnios group (22/30, 77.3% vs 11/30, 36.7%, P<0.05). Most of the TXA(2)R-positive cases in the oligohydramnios group showed strong positivities of TXA(2)R.

CONCLUSIONAbnormal elevation of TXA(2) concentration in the umbilical cord blood and the TXA(2)/PGI(2) imbalance are responsible for the development of oligohydramnios.

Adult ; Alprostadil ; analogs & derivatives ; blood ; Epoprostenol ; blood ; Female ; Fetal Blood ; chemistry ; Humans ; Oligohydramnios ; metabolism ; Placenta ; chemistry ; Pregnancy ; Radioimmunoassay ; Receptors, Thromboxane A2, Prostaglandin H2 ; chemistry ; Thromboxane A2 ; blood

OBJECTIVETo study ingredients penetrable through placental barrier by administering pregnant rats with Scutellaria Radix extract using HPLC-MS.

METHODRats in early, middle and late pregnancy were intragastrically administered with Scutellaria Radix extract for 5 days. Maternal plasma and embryonic tissues were obtained at 1.5, 12 h after the final administration of Scutellaria Radix extract to determine ingredients of biological specimens.

RESULTUnder the optimum experimental conditions, seven compounds were detected in all pregnant rat plasma, specifically including baicalin, wogonoside, baicalein, wogonin and oroxylin A. The seven compounds were also discovered in embryonic tissues of rats in early pregnancy, including the five detected compounds. But they were not detected in embryonic tissues of rats in middle pregnancy, and six compounds except baicalein were detected embryonic tissues of rats in late pregnancy.

CONCLUSIONIngredients contained in Scutellaria Radix are detected in pregnant rats at different stages, except those in middle pregnancy, indicating a potential in utero exposure in case of oral administration of Scutellaria Radix pregnancy. Therefore, a study of embryotoxicity shall be continued to evaluate the safety of Scutellaria Radix extract.

Animals ; Drugs, Chinese Herbal ; adverse effects ; chemistry ; pharmacokinetics ; Female ; Placenta ; metabolism ; Plant Extracts ; adverse effects ; chemistry ; pharmacokinetics ; Pregnancy ; Rats ; Scutellaria baicalensis ; chemistry

OBJECTIVETo test the possibility that certain efficient substrates for lipoxygenase (LOX) produce shuttle oxidants that stimulate the generation of reactive oxygen species from other chemicals.

METHODSMetabolic interactions were conducted in vitro between chlorpromazine (BZ) or other phenothiazines and benzidine or other xenobiotics mediated by soybean lipoxygenase (SLO) or purified human term placental lipoxygenase (HTPLO) in the presence of hydrogen peroxide, and the rates of xenobiotics oxidation were measured by spectroscopic analysis.

RESULTSChlorpromazine cation radical (CPZ(*+)) generated by lipoxygenase triggered a rapid oxidation of benzidine to benzidine diimine cation. Under the experimental conditions used, the metabolic interaction resulted in a 42-fold greater stimulation than BZ alone in the rate of BZ oxidation. The magnitude of stimulation of benzidine oxidation exhibited a dependence on the pH of the reaction medium, amount of the enzyme, and concentration of chlorpromazine, BZ, and hydrogen peroxide. A number of other phenothiazines were also found to stimulate BZ oxidation, albeit to a lesser degree. Chlorpromazine cation radical stimulated the oxidation of all six other xenobiotics tested. The highest stimulation (94-fold) was noted in the oxidation of tetramethyl phenylenediamine to Wursters blue radical, while the lowest stimulatory response (2-fold) was observed in guaiacol. Preliminary data showed that purified HTPLO also displayed a similar stimulatory response to benzidine oxidation in the presence of chlorpromazine.

CONCLUSIONBoth soybean lipoxygenase and purified human term placental lipoxygenase can mediate stimulatory response to the oxidation of benzidine and other xenobiotics in the presence of phenothiazines.

Benzidines ; metabolism ; Chlorpromazine ; chemistry ; Drug Interactions ; Female ; Humans ; Lipoxygenase ; isolation & purification ; pharmacology ; Oxidation-Reduction ; drug effects ; Phenothiazines ; pharmacology ; Placenta ; enzymology ; Pregnancy ; Xenobiotics ; metabolism

ATP-Binding Cassette, Sub-Family B, Member 1 ; chemistry ; genetics ; physiology ; Animals ; Blood-Brain Barrier ; metabolism ; Drug Interactions ; Humans ; Intestinal Absorption ; Pharmacogenetics ; Placenta ; metabolism ; Polymorphism, Genetic

The sequestration of infected erythrocytes in the placenta can activate the syncytiotrophoblast to release cytokines that affect the micro-environment and influence the delivery of nutrients and oxygen to fetus. The high level of IL-10 has been reported in the intervillous space and could prevent the pathological effects. There is still no data of Th17 involvement in the pathogenesis of placental malaria. This study was conducted to reveal the influence of placental IL-17 and IL-10 levels on fetal weights in malaria placenta. Seventeen pregnant BALB/C mice were divided into control (8 pregnant mice) and treatment group (9 pregnant mice infected by Plasmodium berghei). Placental specimens stained with hematoxylin and eosin were examined to determine the level of cytoadherence by counting the infected erythrocytes in the intervillous space of placenta. Levels of IL-17 and IL-10 in the placenta were measured using ELISA. All fetuses were weighed by analytical balance. Statistical analysis using Structural Equation Modeling showed that cytoadherence caused an increased level of placental IL-17 and a decreased level of placental IL-10. Cytoadherence also caused low fetal weight. The increased level of placental IL-17 caused low fetal weight, and interestingly low fetal weight was caused by a decrease of placental IL-10. It can be concluded that low fetal weight in placental malaria is directly caused by sequestration of the parasites and indirectly by the local imbalance of IL-17 and IL-10 levels.
Animals ; Female ; *Fetal Weight ; Humans ; Interleukin-10/*analysis/metabolism ; Interleukin-17/*analysis/metabolism ; Malaria/*metabolism/parasitology/physiopathology ; Male ; Mice ; Mice, Inbred BALB C ; Placenta/*chemistry/metabolism ; Plasmodium berghei/*physiology ; Pregnancy ; Pregnancy Complications, Parasitic/*metabolism/parasitology/physiopathology

OBJECTIVETo investigate the molecular mechanisms of effective ingredients of Astragalus-Salvia compound (ASC) in increasing placental blood supply, to provide thought for establishing an effective prevention and treatment of insufficient placental blood supply caused complications of pregnancy.

METHODSThe effective ingredients of ASC was isolated and abstracted and the rat model of inhibited nitric oxide synthesis was established by using L-Arginine methyl ester (L-NAME), a nitric oxide synthase inhibitor. The mRNA expression of vascular endothelial growth factor (VEGF) in placental trophocyte and primordial micrangial endothelial cell was determined by using in situ hybridization at 12th day of pregnancy.

RESULTSThe VEGF mRNA expression was significantly lower in nitric oxide synthesis inhibited model rats than that in rats non-modeled, or in model rats treated by ASC abstracts or nitroglycerin (P < 0.05, P < 0.01).

CONCLUSIONEffective ingredients of ASC may improve VEGF mRNA expression in placental trophocyte and vascular endothelial cell in early pregnancy, and thus be favorable for placental vascular net formation, trophocyte infiltration and mother-placental circulation constitution for increasing placental blood supply to prevent and treat the insufficient placental blood supply caused diseases such as pregnancy-induced hypertension syndrome.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Endothelial Cells ; cytology ; Female ; Male ; Nitric Oxide ; biosynthesis ; Placenta ; cytology ; metabolism ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Trophoblasts ; metabolism ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics

OBJECTIVETo clone and identify the gene encoding human ubiquitin binding enzyme 2 and study its expression pattern.

METHODSAccording to the sequence of human EST, which is highly homologous to the mouse ubiquitin binding/conjugating enzyme (E2), primers were synthesized to screen the human fetal brain cDNA library. The gene was analyzed by bioinformatics technique and its expression pattern was studied by using multiple-tissue Northern blot.

RESULTSTwo cDNA clones encoding human ubiquitin conjugating enzyme have been isolated and identified. Both containing the ubiquitin conjugating domain, the 2 cDNA clones are 88% identical in amino acid sequences and splicing isoforms to each other only with an exon excised to form the short sequence. They belong to a highly conserved and widely expressed E2 enzyme family. Northern blot shows that they are expressed exclusively in adult human heart, placenta, and pancreas but no transcripts can be detected in brain, lung, liver, skeletal muscle or kidney.

CONCLUSIONSThe gene encoding human ubiquitin binding enzyme is expressed under temporal control. As a key enzyme in the degradation of proteins, ubiquitin conjugating enzymes play a central role in the expression regulation on the level of post-translation.

Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Female ; Humans ; Mice ; Molecular Sequence Data ; Myocardium ; metabolism ; Pancreas ; metabolism ; Placenta ; metabolism ; Rats ; Sequence Alignment ; Sequence Analysis, DNA ; Ubiquitin ; genetics ; Ubiquitin-Conjugating Enzymes ; biosynthesis ; chemistry ; genetics

While human induced pluripotent stem cells (hiPSCs) have promising applications in regenerative medicine, most of the hiPSC lines available today are not suitable for clinical applications due to contamination with nonhuman materials, such as sialic acid, and potential pathogens from animal-product-containing cell culture systems. Although several xeno-free cell culture systems have been established recently, their use of human fibroblasts as feeders reduces the clinical potential of hiPSCs due to batch-to-batch variation in the feeders and time-consuming preparation processes. In this study, we have developed a xeno-free and feeder-cell-free human embryonic stem cell (hESC)/hiPSC culture system using human plasma and human placenta extracts. The system maintains the self-renewing capacity and pluripotency of hESCs for more than 40 passages. Human iPSCs were also derived from human dermal fibroblasts using this culture system by overexpressing three transcription factors-Oct4, Sox2 and Nanog. The culture system developed here is inexpensive and suitable for large scale production.
Cell Culture Techniques ; methods ; Cell Differentiation ; Cellular Reprogramming ; Culture Media ; Extracellular Matrix Proteins ; isolation & purification ; Female ; Fibroblasts ; cytology ; Humans ; Lentivirus ; genetics ; Placenta ; chemistry ; Pluripotent Stem Cells ; cytology ; metabolism ; Pregnancy ; Sodium Chloride ; chemistry ; Transcription Factors ; genetics

OBJECTIVE: The purpose of this study was to determine the role of Ureaplasma urealyticum-derived lipid-associated membrane proteins (LAMPs) in the host innate immune system, specifically their effect on Toll-like receptors (TLRs). METHODS: LAMPs were derived from U. urealyticum strains, and human amniotic epithelial cells (HAECs) were isolated from healthy full-term placentas. Cytokine concentrations were determined by enzyme-linked immunosorbent assay (ELISA) and TLR2 mRNA by real-time PCR. Expression of TLR2 was confirmed by Western blotting and immunohistochemistry. RESULTS: LAMPs induced HAECs to produce inflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor (TNF)-α. Cytokine production was reduced after blocking TLR2 using TLR2 inhibitor (anti-hTLR2-IgA). CONCLUSIONS LAMPs isolated from U. urealyticum induced TLR2-dependent up-regulation of inflammatory genes and cytokines in HAECs.
Amnion/cytology* ; Amniotic Fluid/cytology* ; Cytokines/metabolism* ; Dose-Response Relationship, Drug ; Epithelial Cells/metabolism* ; Female ; Humans ; Inflammation ; Interleukin-6/metabolism* ; Interleukin-8/metabolism* ; Lipids/chemistry* ; Lipopolysaccharides/metabolism* ; Membrane Proteins/metabolism* ; Placenta/metabolism* ; Pregnancy ; Toll-Like Receptor 2/metabolism* ; Tumor Necrosis Factor-alpha/metabolism* ; Up-Regulation ; Ureaplasma urealyticum/metabolism*