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

C-Terminal carboxyl methylation of a human placental 23 kDa protein catalyzed by membrane-associated methyltransferase has been investigated. The 23 kDa protein substrate methylated was partially purified by DEAE-Sephacel, hydroxyapatite and Sephadex G-100 gel filtration chromatographies. The substrate protein was eluted on Sephadex G-100 gel filtration chromatography as a protein of about 29 kDa. In the absence of Mg2+, the methylation was stimulated by guanine nucleotides (GTP, GDP and GTPgammaS), but in the presence of Mg2+, only GTPgammaS stimulated the methylation which was similar to the effect on the G25K/rhoGDI complex. AFC, an inhibitor of C-terminal carboxyl methylation, inhibited the methylation of human placental 23 kDa protein. These results suggests that the substrate is a small G protein different from the G25K and is methylated on C-terminal isoprenylated cysteine residue. This was also confirmed by vapor phase analysis. The methylated substrate protein was redistributed to membrane after in vitro methylation, suggesting that the methylation of this protein is important for the redistribution of the 23 kDa small G protein for its putative role in intracellular signaling.
Cysteine/metabolism* ; Female ; GTP-Binding Proteins/metabolism* ; Guanine Nucleotides/pharmacology ; Human ; Methylation ; Placenta/metabolism* ; Placenta/enzymology ; Pregnancy ; Pregnancy Proteins/metabolism* ; Protein Methyltransferases/metabolism*

OBJECTIVE: To investigate the expression of Calponin-1 and Transgenlin in the uterine smooth muscles during normal labor on-sets, and to evaluate their effect on initiating the normal labor. METHODS: A total of 14 uterine bodies and lower segments of human pregnancy were divided to a non-labor group (NIL) and a labor group(IL). Immunohistochemical technology and Western blot were used to determine the expression of Calponin-1 and Transgelin in the 2 groups. RESULTS: Immunohistochemical detection and Western blot showed that Calponin-1 protein in the uterine smooth muscle tissue of the body and the lower uterine segment of smooth muscle tissues had significant difference (P<0.05). The expression of Transgelin in the uterine body smooth muscle tissue in the IL was higher than that in the NIL(P<0.05). In the lower uterine segments of the smooth muscle, the expression of Transgelin was not significantly different in the 2 groups (P>0.05). CONCLUSION Calponin-1 of the uterine smooth muscle and Transgelin of the uterine body smooth muscle may involve in the regulation of uterine smooth muscle contractility, which is closely related to child birth launch.
Adult ; Calcium-Binding Proteins ; genetics ; metabolism ; Female ; Humans ; Labor Onset ; metabolism ; Microfilament Proteins ; genetics ; metabolism ; Muscle Proteins ; genetics ; metabolism ; Myometrium ; metabolism ; Pregnancy ; Uterine Contraction ; metabolism

OBJECTIVE: To determine the apoptosis in placenta tissues of patients with hypertensive disorder complicating pregnancy and its relationship with Bcl-2, TGFbeta1, and to explore the etiology of hypertensive disorder complicating pregnancy. METHODS: Forty-five placenta samples were obtained from pregnancies with hypertensive disorder (15 gestational hypertension, 15 mild preeclampsia, and 15 severe preeclampsia) and 45 normal placenta tissues were enrolled from the third-trimester pregnancies. Immunohistochemistry (SP method) was used to study the expression of Bcl-2 and TGFbeta1 in human trophoblasts. Terminal deoxynucleotidyl transferase-dUTP nick end-labeling (TUNEL) was used to quantify the incidence of apoptosis in human trophoblasts. RESULTS: The apoptosis rate and TGFbeta1 expression in hypertensive disorder complicating pregnancy group was higher than that in the control group, but the Bcl-2 expression was significantly lower than the control group (all Ps<0.01). With the aggravation of this illness, the apoptosis rate and TGFbeta1 expression in the gestational hypertension group, mild preeclampsia group, and severe preeclampsia tended to be increasing, but the Bcl-2 expression was decreasing (P<0.001). The apoptosis of placenta villi and TGFbeta1 expression were positively correlated in the severe preeclampsia group and mild preeclampsia group,but the apoptosis of placenta villi and Bcl-2 were negatively correlated (all Ps<0.05). TGFbeta1 and Bcl-2 expressions in the severe preeclampsia group and mild preeclampsia group were negatively correlated (P<0.05). CONCLUSION Apoptosis of the placental trophoblasts of pregnancies with hypertensive disorder is evidently enhanced. The TGFbeta1 expression increases and the Bcl-2 expression decreases. The imbalance between TGFbeta1 and Bcl-2 expression may induce the hypertensive disorder.
Adult ; Apoptosis ; Female ; Humans ; Hypertension, Pregnancy-Induced ; metabolism ; Placenta ; cytology ; metabolism ; Pregnancy ; Pregnancy Trimester, Third ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Transforming Growth Factor beta1 ; metabolism

PURPOSE: Recently, COMMD1 has been identified as a novel interactor and regulator of hypoxia-inducible factor-1 and nuclear factor kappa B transcriptional activity. The goal of this study was to determine the difference of COMMD1 expression in the placentas of women with normal and preeclamptic (PE) pregnancies. MATERIALS AND METHODS: Immnoperoxidase and immunofluorescent staining for COMMD1 was performed on nine normal and nine severe PE placental tissues, and COMMD1 mRNA expression was quantified by quantitative reverse transcription polymerase chain reaction. RESULTS: The expression of mRNA of COMMD1 was significantly higher in the study group than in the control group. The immunoreactivity was higher especially in the syncytiotrophoblast of PE placentas than in the control group. CONCLUSION: This study demonstrated increased placental COMMD1 expression in women with severe preeclampsia compared to that found in women with normal pregnancies, and this finding might contribute to a better understanding of the pathophysiology of preeclampsia.
Adaptor Proteins, Signal Transducing/genetics/isolation & purification/*metabolism ; Adult ; Female ; Humans ; Placenta/*metabolism ; Pre-Eclampsia/*metabolism ; Pregnancy ; RNA, Messenger/metabolism

OBJECTIVETo investigate lead distribution and the change of 78 000 glucose regulated protein (GRP78) in various organs of weaned rats challenged with low-level maternal origin lead.

METHODSMale littermates, bred from the female Fisher 344 rats gavaged with lead acetate or sodium acetate (1 ml of 10 mg/ml per day per animal) with male Fisher 344 rats without lead treatment, were divided into 4 groups including control (group A), gestation plus lactation (group B), gestation only (group C), and lactation only (group D). Each group had 6 litters. These littermates were weaned and terminated at postnatal day 21. Lead contents and GRP78 levels in various organs of these littermates were determined by atomic absorbance spectrometry (AAS) and Western blotting analysis, respectively.

RESULTSMaternal lead was observed to transfer to littermates through gestation and lactation. Concentrations of littermate blood lead in groups A to D were (0.0010+/-0.0010), (0.1420+/-0.0190), (0.0250+/-0.0040), and (0.1490+/-0.0160) microg/ml, respectively. Concentrations of littermate brain lead in groups A to D were (0.0005+/-0.0005), (0.1120+/-0.0130), (0.0125+/-0.0042), and (0.0700+/-0.0058) microg/g, respectively. Concentrations of littermate kidney lead in groups A to D were (0.0050+/-0.0050), (1.0400+/-0.1000), (0.1040+/-0.0330), and (0.9920+/-0.0850) microg/g, respectively. Concentrations of littermate liver lead in groups A to D were (0.0030+/-0.0050), (0.3600+/-0.0550), (0.0567+/-0.0126), and (0.3030+/-0.0310) microg/g, respectively. Blood, brain, kidney and liver lead concentrations in groups B and D were significantly higher than those in group C and differences were 5-10 folds. Arbitrary units of littermate leukocytic GRP78 concentration normalized with actin protein in groups A to D were 1.000+/-0.038, 1.180+/-0.060, 0.998+/-0.109, and 1.290+/-0.110, respectively. Arbitrary units of littermate brain GRP78 concentration normalized with actin protein level in groups A to D were 0.996+/-0.128, 0.922+/-0.246, 1.150+/-0.170, and 0.750+/-0.126, respectively.

CONCLUSIONLead in maternal bodies could be transferred to litter bodies through gestation and lactation and distributed in various organs. Lead might also changed GRP78 expression in leukocytes.

Animals ; Animals, Newborn ; Brain ; metabolism ; Female ; HSP70 Heat-Shock Proteins ; metabolism ; Kidney ; chemistry ; Lead ; metabolism ; Leukocytes ; metabolism ; Liver ; chemistry ; Male ; Membrane Proteins ; metabolism ; Pregnancy ; Rats ; Rats, Inbred F344

OBJECTIVE: To study the expression characteristics of NeuN (neuron-specific nuclear protein/neuronal nuclei) in the development of mice olfactory bulb and epithelium. METHOD: The expression of NeuN in the heads of E9.5, E11.5, E14.5, E17.5, newborn and adult mice were examined by immunofluorescence method. RESULT: NeuN was expressed in mature neural cells at the margin area of olfactory bulb of new natal mice while the layers of olfactory bulb could not be distinguished. Mature neural cells expressing NeuN aggregated in the granule cells layer, when layers of olfactory bulb of adult mice were clearly formed. The expression of NeuN could not be observed in bipolar olfactory receptor neurons (ORN) in olfactory epithelium. CONCLUSION It is usually believed that NeuN is expressed in most mature neurons. However, during the development of olfactory bulb and epithelium, NeuN is expressed only in mature neural cells migrating from margin area to the inner granule cells layer of olfactory bulb. It is proposed that the migration of granule cells in olfactory bulb might be related to the formation of smell pattern.
Animals ; Animals, Newborn ; Female ; Mice ; Nerve Tissue Proteins ; metabolism ; Neurons ; metabolism ; Nuclear Proteins ; metabolism ; Olfactory Bulb ; embryology ; growth & development ; metabolism ; Olfactory Mucosa ; embryology ; growth & development ; metabolism ; Pregnancy

OBJECTIVETo detect the expression profiles of suppressor of cytokine signaling 3 (SOCS3), interleukin (IL)-10, and tumor necrosis factor-alpha (TNF-a) in the placenta of women with intrahepatic cholestasis of pregnancy (ICP) and determine the clinical significance of the differential expressions.

METHODSPlacentas were collected from 37 ICP gravidas who delivered through cesarean section at the First Teaching Hospital of Xingjiang Medical University from October 2010 to May 2011 and from 35 healthy pregnant women (controls). SOCS3, TNF-a, and IL-10 protein levels were detected by immunoblotting and the Envision immunohistochemical method.

RESULTSTNF-a and IL-10 expression was detected in placentas of both groups, and was present mainly in the cytoplasm of trophoeytes. IL-10 expression was obviously lower in the ICP placentas than in the control placentas; meanwhile, TNF-a expression was obviously higher than in the control placentas (Z=-2.63, P less than 0.01). SOCS3 protein was significantly more abundant in the control placentas than in the ICP placentas. Furthermore, SOCS3 and IL-10 placental expressions were positively correlated (r=0.494, P less than 0.01), but there was a negative correlation between SOCS3 and TNF-a placental expressions (r=-0.472, P less than 0.01).

CONCLUSIONIn ICP, an increase of the type 1 cytokine, TNF-a, is associated with decreases of the type 2 cytokine, IL-10, and of SOCS3, which may reduce the secretion of IL-10. Furthermore, SOCS3 may contribute to ICP pathogenesis by modulating the Th1/Th2 cytokine balance.

Adult ; Case-Control Studies ; Cholestasis, Intrahepatic ; metabolism ; pathology ; Female ; Humans ; Interleukin-10 ; metabolism ; Placenta ; metabolism ; Pregnancy ; Pregnancy Complications ; metabolism ; pathology ; Pregnancy Trimester, Third ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Young Adult

Teratozoospermia is a rare disease associated with male infertility. Several recurrent genetic mutations have been reported to be associated with abnormal sperm morphology, but the genetic basis of tapered-head sperm is not well understood. In this study, whole-exome sequencing (WES) identified a homozygous WD repeat domain 12 (WDR12; p.Ser162Ala/c.484T>G) variant in an infertile patient with tapered-head spermatozoa from a consanguineous Chinese family. Bioinformatic analysis predicted this mutation to be a pathogenic variant. To verify the effect of this variant, we analyzed WDR12 protein expression in spermatozoa of the patient and a control individual, as well as in the 293T cell line, by Western blot analysis, and found that WDR12 expression was significantly downregulated. To understand the role of normal WDR12, we evaluated its mRNA and protein expression in mice at different ages. We observed that WDR12 expression was increased in pachytene spermatocytes, with intense staining visible in round spermatid nuclei. Based on these results, the data suggest that the rare biallelic pathogenic missense variant (p.Ser162Ala/c.484T>G) in the WDR12 gene is associated with tapered-head spermatozoa. In addition, after intracytoplasmic sperm injection (ICSI), a successful pregnancy was achieved. This finding indicates that infertility associated with this WDR12 homozygous mutation can be overcome by ICSI. The present results may provide novel insights into understanding the molecular mechanisms of male infertility.
Humans ; Pregnancy ; Female ; Male ; Animals ; Mice ; Teratozoospermia/pathology* ; Semen/metabolism* ; Infertility, Male/metabolism* ; Spermatozoa/metabolism* ; Mutation ; RNA-Binding Proteins/metabolism* ; Cell Cycle Proteins/genetics*

OBJECTIVEThe aim of this study was to investigate the ability of Pref-1+ adipocyte progenitor cells to mobilize into mesenteric lymph nodes (MLNs) and the dynamic expression of related chemokines during the development of rat MLNs.

METHODSImmunohistochemical analyses were used to detect the expression of Pref-1 and related chemokines. Transmission electron microscopy (TEM) was used to observe the changes in ultrastructure of MLNs.

RESULTSCells containing lipid droplets were found in all rat MLNs at embryonic day (E) 18.5, 2 and 6 weeks (w) after birth, and they were similar to fibroblastic reticular cells (FRCs) or follicular dendritic cells (FDCs) under TEM. Pref-1+ adipocyte progenitor cells were found in all MLNs. The expression level of Pref-1 was significantly increased at 2 w after birth and decreased at 6 w after birth. The tendency of Cxcl12 expression was consistent with that of Pref-1 and was positively correlated with the expression of Pref-1 (P < 0.01; r = 0.897). At E18.5, Cxcl13, and Ccr7 were significantly expressed in the MLN anlage, but the expression level of Ccl21 was low. The expression level of Cxcl13, Ccr7, and Ccl21 in MLN were significantly increased at 2 w after birth (P < 0.05), while the expression of Ccr7 and Ccl21 were significantly decreased at 6 w after birth (P < 0.05).

CONCLUSIONAdipocyte progenitor cells are involved in the rat MLNs development through differentiation into FRC and FDC. The expression of the relevant chemokines during the development of MLNs is dynamic and may be related to the maintenance of lymph nodes self-balance state.

Animals ; Chemokines ; genetics ; metabolism ; Female ; Gene Expression Regulation, Developmental ; physiology ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Lymph Nodes ; embryology ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Mesentery ; embryology ; Pregnancy ; Rats