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

OBJECTIVETo investigate the expression of TM4SF9 in the villi of early pregnancy, hydatidiform mole, invasive hydatidiform mole and chorionic carcinoma tissue.

METHODSImmunohistochemistry was used to detect the expression of TM4SF9 in normal villi of early pregnancy, hydatidiform mole, invasive hydatidiform mole and chorionic carcinoma tissues.

RESULTSTM4SF9 was expressed in the cytotroblasts but not in the syncytiotrophoblast of normal villi. The intensity of TM4SF9 expression increased in the order of normal villi, hydatidiform mole, invasive hydatidiform mole and chorionic carcinoma, with strong positivity rates of 0, 10%, 36.4% and 100%, respectively, showing significant differences between the samples (P<0.001).

CONCLUSIONTM4SF9 expression in the trophoblasts may relate to their invasiveness and play an important role in the metastasis of trophoblastic tumor.

Adult ; Choriocarcinoma ; metabolism ; Chorionic Villi ; metabolism ; Female ; Humans ; Hydatidiform Mole ; metabolism ; Hydatidiform Mole, Invasive ; metabolism ; Immunohistochemistry ; Membrane Proteins ; biosynthesis ; Pregnancy ; Tetraspanins ; Trophoblastic Neoplasms ; metabolism ; Trophoblasts ; metabolism ; Uterine Neoplasms ; metabolism

OBJECTIVETo discuss the effect of Yisui Shengxue granules on expression of alpha-hemoglobin stabilizing protein (AHSP) mRNA in different developmental stages mice.

METHODThe total RNAs were extracted from the bone marrow karyocyte of normal adult mice and the karyocyte of fetus liver and fetus spleen in pregnanted mice (pregnanted 21 days) and fetal mice (pregnanted 14 days). The expression level of AHSP mRNA in different developmental stages mice interfered with Yisui Shengxue granules was measured by real-time PCR.

RESULTThe intervention of Yisui Shengxue granules could significantly up-regulated the expression levels of AHSP mRNA in normal adult mice.

CONCLUSIONThe result revealed that one of possible molecular mechanism of the effects caused by Yisui Shengxue granules is that it can promote the AHSP gene expression, reduce the free a-globin deposit, then prevent the poison to erythrocyte and decrease the haemolysis.

Animals ; Blood Proteins ; genetics ; Bone Marrow Cells ; cytology ; drug effects ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Erythrocytes ; cytology ; drug effects ; metabolism ; Female ; Gene Expression Regulation, Developmental ; drug effects ; Liver ; cytology ; embryology ; metabolism ; Male ; Mice ; Molecular Chaperones ; genetics ; Plants, Medicinal ; chemistry ; Pregnancy ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Spleen ; cytology ; embryology ; metabolism ; Up-Regulation ; drug effects ; genetics

OBJECTIVETo construct adeno-associated virus (AAV) expression system for transforming growth factor beta3 (TGFbeta3 ) and detect its biological effect on proteoglycan synthesis of the earlier and later dedifferentiated rabbit lumbar disc nucleus pulpous (NP) cells, which was compared with that of adenovirus (AV) expression system for TGFbeta1.

METHODSTGFbeta3 gene was obtained using PCR. Its upstream contained restriction enzyme site Kpn I, and its downstream contained restriction enzyme site Sal I. Using the restriction enzyme sites of PCR product of TGFbeta3 and the corresponding multiple cloning site (MCS) in plasmid AAV, TGFbeta3 was subcloned into AAV. The recombinant plasmid AAV-TGFbeta3 was transfected into H293 cells with Lipofectamine 2000, and the expression of TGFbeta3 gene was detected using immunofluorescent analysis. After AAV-TGFbeta3 virus particle with infectious activity was packaged, TGFbeta3 expression in NP cells was detected by immunoblotting, and its biological effect on proteoglycan synthesis was detected by antonopulos method and compared with that of AV-TGFbeta1 in the earlier and later dedifferentiated NP cells.

RESULTSFor the earlier dedifferentiated NP cells, AAV-TGFbeta3 slowly and stably enhanced proteoglycan synthesis, but AV-TGFbeta1 rapidly and transiently enhanced its synthesis. For the later dedifferentiated NP cells, AAV-TGFbeta3 stably enhanced proteoglycan synthesis, but AV-TGFbeta1 inhibited its synthesis.

CONCLUSIONAAV expression system can mediate TGFbeta3 gene to be expressed stably, and AAV-TGFbeta3 can enhance proteoglycan synthesis of the earlier and later dedifferentiated NP cells.

Animals ; Cell Line ; DNA, Recombinant ; genetics ; Dependovirus ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc ; cytology ; Placenta ; cytology ; Plasmids ; genetics ; Polymerase Chain Reaction ; Pregnancy ; Proteoglycans ; biosynthesis ; Rabbits ; Transforming Growth Factor beta1 ; genetics ; Transforming Growth Factor beta3 ; genetics ; Viral Proteins ; biosynthesis

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

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

In the present study, whether the ADAM-8, -9, -10, -12, -15, -17, and ADAMTS-1 proteins might play a role in mouse uterus during periimplantation period was investigated. Immunoblotting analyses demonstrated that all ADAM proteins consistently appeared throughout days 1 to 8 of pregnancy but with a variation depending on the species of ADAM gene, the progression of pregnancy, and the site of the uterus. Immunohistochemical analyses indicated that ADAM proteins were localized in the luminal or glandular epithelial layers with a varying intensity depending on the species of ADAM and the progression of pregnancy. Particularly ADAM-8, -12, and -15, were predominantly located in the implantation site of the uterine tissues, whereas little or no protein was localized in the interimplantation site. Based upon these observations, it is suggested that the ADAMs might play an important role in the remodeling of the mouse uterus during the periimplantation period.
Uterus/*metabolism ; Time Factors ; Pregnancy ; Mice, Inbred ICR ; Mice ; Immunohistochemistry ; Immunoblotting ; *Gene Expression Regulation ; Female ; *Estrous Cycle ; Embryonic Development ; *Embryo Implantation ; Animals ; ADAM Proteins/*biosynthesis/*physiology

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

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

OBJECTIVETo determine the threshold of millimeter wave irradiation for fetal injury in mice and the mechanism of decrease of learning and memory function in their offsprings and to verify whether the millimeter wave has the non-thermal effect.

METHODSPregnant mice were irradiated by millimeter wave with frequencies of 37.4, 42.2, 53.0 and 60.0 GHz at power densities of 1, 3, 5, 8 mW/cm(2) for two hours daily from the 6th to 15th day of their gestation. Learning and memory functions of their offsprings were tested by a Y-type electric maze. c-Fos protein expression level in hippocampus of their offsprings was determined with immunohistochemistry 0, 30, 60, 90 and 120 minutes after the offsprings were trained respectively.

RESULTSThe minimal power density of millimeter wave for the decrease in learning and memory function and decrease of c-Fos protein expression level in hippocampus of their offsprings caused by 37.4, 42.2 GHz and 53.0, 60.0 GHz was 5 and 3 mW/cm(2). Severity of injury for learning and memory in offsprings caused by irradiation increased with the power density of millimeter wave. The millimeter wave did not cause increase of the body temperature of the pregnant mice.

CONCLUSIONThe threshold of millimeter wave with 37.4, 42.2 GHz, and 53.0, 60.0 GHz causing fetal injury in mice is 5 and 3 mW/cm(2) respectively. The decrease in learning and memory functions in offspring mice is related with decrease of c-Fos protein expression level in hippocampus. Millimeter wave has the non-thermal effects.

Animals ; Dose-Response Relationship, Radiation ; Female ; Hippocampus ; metabolism ; radiation effects ; Immunohistochemistry ; Learning ; radiation effects ; Memory ; radiation effects ; Mice ; Mice, Inbred BALB C ; Microwaves ; adverse effects ; Pregnancy ; Prenatal Exposure Delayed Effects ; Proto-Oncogene Proteins c-fos ; biosynthesis