To investigate the expressions of placental growth factor (PLGF) in placenta with hypertensive disorders of pregnancy (HDP), 45 women with HDP and 20 normally pregnant women were studied. Among 45 women with HDP, there were 23 cases of severe preeclampsia and one case of eclampsia. The location and level of PLGF proteins was determined by immunohistochemistry and Western blot. The expression of PLGF mRNA in placenta was assessed by reverse transcriptional-polymerase chain reaction (RT-PCR). The results showed that: (1) The distribution of PLGF in placenta with HDP was similar to normal one, which was mainly in the cytoplasm of villous syncytiotrophoblast and villous stroma; (2) The expression of PLGF protein was significantly decreased in placentas with mild and severe preeclampsia compared to the normal ones (0.3 +/- 0.4 vs 0.6 +/- 0.4, 0.2 +/- 0.5 vs 0.6 +/- 0.4, P < 0.01). There were no differences between the gestational hypertension placenta and normal one (0.5 +/- 0.6 vs 0.6 +/- 0.4, P > 0.05); (3) The transcription levels of the PLGF mRNA in placentas with preeclampsia were significantly lower than in normal groups (3.33 +/- 0.39 vs 4.87 +/- 0.60, 1.97 +/- 0.29 vs 4.87 +/- 0.60, P < 0.01), and no differences were found between the gestational hypertension placenta and normal groups. These findings suggest that the abnormal expression of PLGF in placentas is related to the pathogenesis of HDP.
Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy/*metabolism ; Pregnancy Proteins/*biosynthesis ; Pregnancy Proteins/genetics

The relationship between T cell immunoglobulin domain and mucin domain protein 3 (Tim-3)/Galectin (Gal)-9 pathway and recurrent spontaneous abortion (RSA) was studied. Thirty-one pregnant women with RSA and 27 normal early gravidas were investigated to detect the levels of Tim-3 and Gal-9 in villi and deciduas by Western blotting. Meanwhile, the concentration of interleukin (IL)-4 and IL-12 in peripheral blood plasma was determined by ELISA in 25 healthy fertile non-pregnant controls, the normal early gravidas and pregnant women with RSA mentioned above, respectively. It was found that the relative expression levels of Tim-3 and Gal-9 in villi and deciduas were significantly increased in pregnant women with RSA as compared with those in the normal early gravidas. The concentration of IL-4 in peripheral blood plasma of pregnant women with RSA was lower than that of the normal early gravidas (P<0.05) and healthy fertile non-pregnant controls (P<0.05), but that of IL-2 in pregnant women with RSA was significantly higher than that of the normal early gravidas (P<0.05) and healthy fertile non-pregnant controls (P<0.05). It was suggested that the overexpression of Tim-3/Gal-9 pathway may be related to the pathogenesis of RSA.
Abortion, Spontaneous ; metabolism ; pathology ; Adolescent ; Adult ; Chorionic Villi ; metabolism ; pathology ; Female ; Galectins ; biosynthesis ; Hepatitis A Virus Cellular Receptor 2 ; Humans ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Membrane Proteins ; biosynthesis ; Pregnancy ; Pregnancy Proteins ; biosynthesis ; Up-Regulation

The purpose of this study was to determine whether the levels of soluble fms-like tyrosine kinase-1 (sFlt-1) and placenta growth factor (PlGF) are altered during the second trimester in the plasma of women who subsequently develop preeclampsia. We performed a case-control study to compare the levels of sFlt-1 and PlGF in the preeclamptic (n=46) and normal pregnant women (n=100). The maternal plasma levels of sFlt-1 and PlGF were measured by enzyme-linked immunosorbent assay. The sFlt-1 levels were significantly higher in the preeclamptic women than in normal controls (p<0.001), while the PlGF levels were significantly lower (p<0.001). In normal controls, sFlt-1 levels were positively correlated (r=0.27, p=0.008), whereas, in the preeclamptic women, those were negatively correlated with the PlGF levels (r=-0.423, p=0.005). Furthermore, the log[sFlt-1/PlGF] ratio was significantly higher in the preeclamptic women than in normal controls (p<0.001). The receiver operating characteristic curve revealed a specificity of 78% with a diagnostic sensitivity of 80.4%; the optimal cut-off value of the log[sFlt-1/PlGF] ratio was 1.4 (95% CI 0.756-0.910, p<0.001). Preeclampsia showed a strong association with increased levels of sFlt-1 and decreased levels of PlGF in the second trimester maternal plasma. Accordingly, the sFlt-1/PlGF ratio may provide early prediction of subsequent development of preeclampsia.
Adult ; Biological Markers/metabolism ; Case-Control Studies ; Female ; Humans ; Immunoassay ; Middle Aged ; Placenta/metabolism ; Pre-Eclampsia/*diagnosis/*metabolism ; Pregnancy ; Pregnancy Proteins/*biosynthesis ; Pregnancy Trimester, Second ; ROC Curve ; Sensitivity and Specificity ; Vascular Endothelial Growth Factor Receptor-1/*biosynthesis

Animals ; Electromagnetic Fields ; Embryo, Mammalian ; metabolism ; Female ; Mice ; Pregnancy ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

To study the expression of Dickkopf-1 (DKK-1) in endometrium of pregnant mice during the peri-implantation period and the role of DKK-1 during the embryo implantation in mice. Immunohistochemical technique was employed to determine the location of DKK-1 protein in endometrium, and semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) was utilized to determine the levels of DKK-1 mRNA. Our results showed that the expressions of DKK1 mRNA and protein were higher in experimental groups than in control group (P<0.01) and it increased significantly on day 3 and reached its peak on day 4, and then decreased gradually on day 5-7. The levels of DKK-1 mRNA and protein on day 4 was significantly higher than those of other groups (P<0.01). It is concluded that DKK-1 probably plays an important role in signal transudation of embryo implantation and its high expression indicates the opening of implantation window.
Animals ; Embryo Implantation ; genetics ; Endometrium ; metabolism ; Female ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; genetics ; Male ; Mice ; Pregnancy ; Protein Biosynthesis ; RNA, Messenger ; genetics ; Transcription, Genetic

OBJECTIVETo investigate the expression of transcription regulator LMO4 mRNA in the developing mouse molar and compare the expression pattern of LMO4 with that of Shh signaling molecule.

METHODSWild-type embryos used in this study (E11.5-P1.5) were generated by mating Kun-Ming mice. The expression pattern of LMO4 during organ development was carried on by whole-mount in situ hybridization. The expression patterns of LMO4 and Shh mRNA during molar development were analysed by section in situ hybridization. Immunohistochemical staining of PCNA was carried on by SP method.

RESULTSLMO4 mRNA was widespread at early embryonic stages (E11.5) with positive hybridization signal in the mandibular reason, limb bud, brain, epidermis and somites revealed by whole-mount in situ hybridization. Section in situ hybridization showed that LMO4 was expressed in the tooth bud, the two tips of the enamel organ and the cervical loop from E13.5 to E16.5. While Shh was localized in the enamel knot on E14.5. On E18.5-P1.5, LMO4 transcripts were distributed in the ameloblast and the stratum intermedium. On E13.5-E16.5, the tooth bud cells and the cervical loop cells were PCNA positive. These were the same regions that showed LMO4 mRNA expression.

CONCLUSIONSLMO4 was confined to the dental epithelium and had spatial temporal expression patterns during tooth morphogenesis. The expression patterns of LMO4 and Shh were similar. In early tooth development, LMO4 might regulate cell proliferation. In late tooth development, it might participate in the ameloblast differentiation.

Adaptor Proteins, Signal Transducing ; Animals ; Animals, Newborn ; Female ; Gene Expression Regulation, Developmental ; Homeodomain Proteins ; biosynthesis ; LIM Domain Proteins ; Mice ; Mice, Inbred Strains ; Morphogenesis ; genetics ; Pregnancy ; Tooth Germ ; metabolism ; Transcription Factors ; biosynthesis ; Transcription, Genetic

OBJECTIVEIn this study, we assayed promoter hypermethylation and protein expression of the mismatch repair gene (MMR) hMLH1 and hMSH2 in gestational trophoblastic diseases to understand the significance of MMR promoter methylation and expression in the pathogenesis and malignant transformation of hydatidiform mole.

METHODSDNA was extracted from chorion of early pregnancies, partial hydatidiform moles, complete hydatidiform moles, and invasive moles were over digested by methylation sensitive endonuclease Hpa II. Then the promoters were amplificated by polymerase chain reaction. The protein was detected by immunohistochemistry.

RESULTSIn the normal placenta, neither hMLH1 nor hMSH2 promoter methylation was detected. Expression of hMLH1 and hMSH2 in cytotrophoblasts was strongly positive, and that was negative or weakly positive in syncytiotrophobasts. In all normal chorion, expression of hMLH1 and hMSH2 in cytotrophoblasts was strongly positive. In partial hydatidiform mole and complete hydatidiform mole, the methylation of hMLH1 and hMSH2 promoters was significantly higher than that of early placenta (P < 0.05), and the protein expression in cytotrophoblasts was significantly lower (P < 0.05). In the invasive mole, hMLH1 and hMSH2 promoter methylation were not significantly different as compared with the partial hydatidiform mole and complete hydatidiform mole (P > 0.05). Expression of hMLH1 in the invasive mole (54.5%, 6/11) was not significantly different as compared with the partial hydatidiform mole and complete hydatidiform mole (P > 0.05). But expression of hMSH2 in the invasive mole (36.4%, 4/11) was weaker than that in complete hydatidiform mole (P = 0.044). Promoter methylation and less expression of hMSH2 had correlations in complete hydatidiform mole or invasive mole.

CONCLUSIONSStrong expressions of hMLH1 and hMSH2 in the cytotrophoblasts of normal placenta may keep the genome stability. Promoter methylation and down-regulation of hMLH1 and hMSH2 are probably involved in the pathogenesis of hydatidiform mole.

Adaptor Proteins, Signal Transducing ; Adult ; Base Pair Mismatch ; genetics ; Carrier Proteins ; DNA Methylation ; DNA Repair ; DNA-Binding Proteins ; biosynthesis ; Female ; Humans ; Hydatidiform Mole ; genetics ; pathology ; Hydatidiform Mole, Invasive ; genetics ; pathology ; Middle Aged ; MutL Protein Homolog 1 ; MutS Homolog 2 Protein ; Neoplasm Proteins ; biosynthesis ; Nuclear Proteins ; Pregnancy ; Promoter Regions, Genetic ; genetics ; Proto-Oncogene Proteins ; biosynthesis ; Uterine Neoplasms ; genetics ; pathology

OBJECTIVETo detect expression of mouse ARL-1 homologous proteins in mouse tissues, and analyze homology, genetic distance and phylogenetic relationship between human aldose reductase like-1 (ARL-1) and mouse homologous proteins.

METHODSHomology of mouse ARL-1 homologous proteins with human ARL-1 was analyzed by software Clustal X 1.8 using GenBank and Swiss-Prot database; genetic distance and phylogenetic relationship between mouse ARL-1 homologous proteins and human ARL-1 were analyzed by software Mega 2.0; mouse tissues were detected by Western blotting using polyclonal antibodies against ARL-1 protein from domestic rabbits.

RESULTSThe amino acid sequence of human ARL-1 was 83%, 82%, 81%, 79%, 70%, 51%, 50% and 45% identical to that of the Chinese hamster ovary reductase (CHO-Red), the mouse fibroblast growth factor-regulated protein (FR-1), rat aldose reductase-like protein (rARLP), the mouse vas deferens protein (MVDP), rat lens aldose reductase (LeAR), delta4-3-ketosteroid-5beta-reductase (5beta-Red), rat aldo-keto reductase protein c (RaK-c) and 3alpha-hydroxysteroid dehydrogenase (3alpha-HSD). Among all the mouse ARL-1 homologous proteins, the genetic distance between CHO-Red and human ARL was the shortest (18.0%, P = 0.023), next was FR-1 (19.1%, P=0.023) and rARLP (19.9%, P = 0.025). From the phylogenetic tree, the protein whose relationship with human ARL-1 was the closest with CHO-Red, next was mouse FR-1, rARLP, MVDP and LeAR. Homologous proteins were found in mouse tissues including vas deferens, testis, bladder and uterus by Western blotting using polyclonal antibodies against ARL-1 protein from domestic rabbits.

CONCLUSIONSCHO-Red has the highest homology, the shortest genetic distance and the closest relationship with human ARL-1, next is FR-1, rARLP, MVDP. The major distribution of mouse ARL-1 homologous proteins is in vas deferens, testis, bladder and uterus, deducing they might be CHO-Red, FR-1, rARLP or MVDP

ADP-Ribosylation Factors ; biosynthesis ; genetics ; Aldehyde Reductase ; biosynthesis ; genetics ; Animals ; Cloning, Molecular ; Cricetinae ; Cricetulus ; Female ; Gene Expression ; Humans ; Male ; Membrane Proteins ; biosynthesis ; genetics ; Mice ; genetics ; Pregnancy ; Sequence Homology, Amino Acid

OBJECTIVETo construct the fusion expression vector of pancreatic-duodenal homeobox gene 1 (PDX-1) fused to green fluorescent protein (GFP) capable of stable expression in fetal rat hepatic stem cells after transfection by electroporation.

METHODSPDX-1 cDNA was amplified from SK900/BLSCRIPT plasmid and cloned into the multiple cloning site of pEGFP-C1 to obtain the recombined plasmid pEGFP-C1-PDX-1. Rat fetal hepatic stem cells were isolated, cultured, identified and transfected with the recombinant vector by electroporation, followed by observation of these cells with fluorescent microscope. The result of transfection was analyzed by RT-PCR and cell growth curve.

RESULTSIdentification by enzyme digestion confirmed successful construction of the recombinant vector. Fetal hepatic stem cells can stably express GFP and PDX-1 for a period of time, and their growth and proliferation was not obviously affected after transfection.

CONCLUSIONThe fusion expression vector of EGFP-PDX-1 is successfully constructed and stably expressed in rat fetal hepatic stem cells, which may facilitate the study of the role of PDX-1 in stem cell differentiation into insulin-producing cells.

Animals ; Electroporation ; Female ; Fetal Stem Cells ; cytology ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Hepatocytes ; cytology ; metabolism ; Homeodomain Proteins ; biosynthesis ; genetics ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Trans-Activators ; biosynthesis ; genetics ; Transfection ; methods

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 micrograms 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.
Apoptosis ; Chorionic Villi ; metabolism ; Decidua ; metabolism ; Female ; Gene Expression Regulation ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Mifepristone ; therapeutic use ; Neoplasm Proteins ; Pregnancy ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Reverse Transcriptase Polymerase Chain Reaction ; Trophoblasts ; metabolism