Localization of chorionic gonadotropin in the albino rat placenta was studied by an immunofluorescent staining method. Intense apple-green fluorescence was observed in the cytoplasm of the trophoblast giant cells of the junctional zone from the 12th to 19th day of pregnancy. The trophospongial cells and glycogen cells of the junctional zone, and decidual cells showed no distinct green fluorescence, but granular green fluorescence wasobserved in the cytoplasm of the trophoblast cell of the labyrinth from the 16th to 18th day of pregnancy. The epithelium of the amnion investing the fetal surface of the placenta and the umbilical cord, which is covered with a single layer of amnion cells, showed no fluorescence. Methyl-green pyronin positive substance (RNA) appeared in relatively large amounts in the cells of yolk sac, labyrinth, junctional zone, and decidua from the 8th day of pregnancy. Albino rat chorionic gonadotropin was localized mostly in the cytoplasm of the trophoblast giant cells in the junctional zone and a smaller amount in the trophoblast cells of the lobyrinth by means of the immunofluorescent technic.
Animals ; Chorionic Gonadotropin/*analysis ; Female ; *Fluorescent Antibody Technique ; Placenta/*analysis ; Rabbits ; Rats

The feasibility of species identification using sequence analysis of the cytochrome B (Cyt B) gene in mitochondrial DNA was investigated. DNA was extracted from nine different animals that could be easily met in our surroundings and Cyt B gene was amplified. Direct sequencing results for the amplified PCR products were compared with each other. Human was also included. Nucleotide sequence of the Cyt B gene for each animals was also compared with the previously known ones registered in nucleotide databases, Genebank. The inter-species sequence variation was high as the percent similarity of each sequences ranged 64.6-83.5%. Compared to this the percent similarity of sequences obtained here were high when compared to the sequences of the same species registered in the database showing relatively low intra-species variation. This data shows that the nucleotide sequences of Cyt B gene in a certain biological materials can be identified at species level. The applicability of this method to the forensic field is also demonstrated by performing a casework; determination of the origin for the placentae which were commercial available as "invogorant". Points about the use of Cyt B gene in forensic field was also reviewed.
Animals ; Base Sequence ; Cytochromes b* ; Cytochromes* ; DNA ; DNA, Mitochondrial ; Humans ; Placenta ; Polymerase Chain Reaction ; Sequence Analysis

The feasibility of species identification using sequence analysis of the cytochrome B (Cyt B) gene in mitochondrial DNA was investigated. DNA was extracted from nine different animals that could be easily met in our surroundings and Cyt B gene was amplified. Direct sequencing results for the amplified PCR products were compared with each other. Human was also included. Nucleotide sequence of the Cyt B gene for each animals was also compared with the previously known ones registered in nucleotide databases, Genebank. The inter-species sequence variation was high as the percent similarity of each sequences ranged 64.6-83.5%. Compared to this the percent similarity of sequences obtained here were high when compared to the sequences of the same species registered in the database showing relatively low intra-species variation. This data shows that the nucleotide sequences of Cyt B gene in a certain biological materials can be identified at species level. The applicability of this method to the forensic field is also demonstrated by performing a casework; determination of the origin for the placentae which were commercial available as "invogorant". Points about the use of Cyt B gene in forensic field was also reviewed.
Animals ; Base Sequence ; Cytochromes b* ; Cytochromes* ; DNA ; DNA, Mitochondrial ; Humans ; Placenta ; Polymerase Chain Reaction ; Sequence Analysis

OBJECTIVE: The purpose of this study was to compare the umbilical artery blood gas analysis and assess the relationship between fetal oxygenation and placenta to birth weight ratios in preeclampsia and small for gestational age. METHODS: We compared the results of umbilical artery blood gas analysis and placenta to birth weight ratio in group of preeclampsia (N=28), group of small for gestational age (N=15), group of large for gestational age (N=15), and controls (N=24). And we also divided all of them into 3 groups by placenta to birth weight ratio at birth, <0.2 (N=25), 0.2-0.25 (N=39) and >0.25 (N=18). We compared umbilical artery gas analysis in each groups. RESULTS: The placenta to birth weight ratio in PE was significantly lower than control group (p<0.05). Umbilical artery pO2 and O2 saturations in each group of preeclampsia and small for gestational age were significantly lower than group of large for gestational age and controls (p<0.05). But we could not find any differences in other umbilical artery blood gas analysis (pH, pCO2, HCO3-). Umbilical artery pO2 and O2 saturations of higher placenta to birth weight ratio were stepwise lower than those of lower placenta to birth weight ratio but, pCO2 of higher placenta to birth weight ratio was stepwise lower than those of lower placenta to birth weight ratio. But there was no significant difference. CONCLUSION: Our data suggested that fetal oxygenation is significant determinant of fetal growth from small for gestational age and preeclampsia. And it may be related to placental implantaton and growth.
Birth Weight* ; Blood Gas Analysis* ; Fetal Development ; Gestational Age* ; Oxygen ; Parturition* ; Placenta* ; Pre-Eclampsia* ; Umbilical Arteries*

OBJECTIVETo investigate the biological characteristics and genetic features of human placenta mesenchymal stem cells (hPA-MSCs) cultured in vitro in order to assess its safety for clinical use.

METHODSThe shapes of the 1st, 3rd, 5th, 7th, 10th, 13th, 17th and 20th generation hPA-MSCs cultured in vitro using serum-free culture medium were observed. Their cell cycle, cell surface markers, and karyotype were analyzed, and relevant genes and cytokines were measured.

RESULTSThe shape of hPA-MSCs has remained as fusiform or short fusiform, and there was no significant change. About 93% of hPA-MSCs cells were in G0/G1 phase and remained stable. No obvious chromosomal translocation, loss or inversion was noted by karyotyping analysis. Cytokines expression level remained stable. Related gene expression level as a whole was on the decline, but the gene expression level of the first five generations showed very slight variations, with genetic characteristics remaining stable.

CONCLUSIONThe hPA-MSCs cultured in vitro with serum-free medium has retained stable in the first five generations.

Cells, Cultured ; Cytokines ; analysis ; Female ; Humans ; Karyotyping ; Mesenchymal Stromal Cells ; physiology ; Placenta ; cytology ; Pregnancy

The present study was aimed to investigate the influence of freeze-drying on the quality of polymerized human placenta hemoglobin (PolyPHb). The PolyPHb solution was freeze-drying under suitable conditions. Hemoglobin concentration, methemoglobin (MetHb) content, UV spectrum, Fe3 content, oxygen-carrying capacity, pH, the average molecular weight and its distribution, circular dichroism, oxygen equilibrium curve and other indicators were measured before and after freeze-drying. The appearance, residual water content, rehydration time of the lyophilized product were also evaluated. The results showed that there was no significant difference on all the indicators measured above, which indicated that freeze-drying process had no effect on the physical and chemical properties of PolyPHb, as well as on its biological activity. Therefore, the properties of PolyPHb were stable during this freeze-drying process and could be preserved after such freeze-drying process.
Blood Substitutes ; Female ; Freeze Drying ; methods ; Hemoglobins ; chemistry ; Humans ; Methemoglobin ; analysis ; Placenta ; chemistry ; Polymerization ; Pregnancy

OBJECTIVE: To study the effects of in vitro fertilization-embryo transfer (IVF-ET) technique on gene expression of focal adhension kinase (FAK) signaling pathway in early placental trophoblast cells, and to explore the effects of IVF-ET technology on the development and function of early placenta. METHODS: We collected 7-8 weeks of gestation placenta tissue as a study group by ultrasound guided reduction of fetal from double embryo transfer under IVF-ET technology. In the control group, placenta tissues were obtained from the spontaneous abortion of natural pregnancy twin 7-8 weeks. Microarray hybridization analysis was performed on the placenta tissue of the two groups using the Affymetrix HG-U133 Plus 2.0 gene chip. Eight differentially expressed genes were identified by real-time quantitative polymerase chain reaction (qRT-PCR), and unsupervised clustering analysis and functional bioinformatics analysis were performed for the differentially expressed genes. RESULTS: Twenty-eight cases of IVF-ET reduced fetal villi and 8 cases of spontaneous abortion villi were collected. A total of 8 placental villi were detected by the gene chip. Compared with the natural pregnancy control group, 32 differentially expressed genes in the placental FAK signaling pathway were expressed in IVF-ET. The differential expression was greater than or equal to 2 times, of which 12 genes were up-regulated and 20 were down-regulated. The qRT-PCR showed that the expression of the 8 genes in FAK signaling pathways of IVF-ET was significantly different from that in the placenta of natural pregnancy, which was consistent with the result of the gene chip detection. The FAK signal pathway gene localization showed that the FAK gene was mainly located in the upstream of the signal pathway in the placenta of IVF-ET. The placental trophoblast cells maintained the FAK signaling pathway function through gene expression compensation. CONCLUSION There are gene expression differences in the FAK signaling pathway between the IVF-ET derived early placenta and the natural pregnancy placenta. The differentially expressed genes are involved in many key functions of the FAK signaling pathway and affect the early development and function of the IVF-ET placenta, while the placental trophoblast cells change gene expression for interference to compensate for IVF-ET technology itself, maintain normal function of the FAK signaling pathway, and satisfy the need for placental and fetal development.
Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Oligonucleotide Array Sequence Analysis ; Placenta ; Pregnancy ; Signal Transduction

PURPOSE: To evaluate a role as over 35 years, maternal serum markers, and a false positive screen for Down syndrome were the predictor of adverse pregnancy outcome. Materials and METHODS: From Mar.1994, through Feb.1996, 5284 women were screened triple test to detect Down syndrome in the second trimester and were delivered Samsung Cheil hospital. The values of each maternal serum markers were measured with radioimmunoassay. And then, the screen positive of Down syndrome was calculated using alpha-software Version 4.0. The adverse outcome of the fetus and the mother included low birth weight(LBW) ( <2500gm), prematurity( <37 gestational weeks), placenta previa, preterm premature rupture of membranes(PPROM), pregnancy induced hypertension(PIH),abruptio placenta, and intrauterine fetal death(IUFD). The predictor markers included over 35years, elevated alpha-fetoprotein (AFP), elevated human chorionic gonadotropin(hCG), lowered unconjugated estriol (uE3), and a false positive screen for Down syndrome. RESULTS: Mean age and mean gestational weeks in the study were 30+/-4.8 years and 17.1 weeks respectively. The adverse pregnancy outcomes were 357 LBW(6.8%), 253 prematurity(4.8%), 108 placenta previa(2.0%), 68 PPROM(1.3%), 66 PIH(1.3%), 24 abruptio placenta(0.5%), and 20 IUFD(0.4%). In univariate analysis, over 35 years was significantly associated with abruptio placenta, prematurity, and placenta previa, elevated MS-AFP( >2.0 MoM) associated with IUFD, LBW, PIH, prematurity, and PPROM , elevated MS-hCG (>3.0 MoM) associated with IUFD, LBW, PIH, prematurity, and placenta previa, lowered uE3 (<0.75) associated with IUFD, abruptio placenta, LBW, and prematurity.(P <0.05). In multivariate logistic regression analysis, IUFD was significantly associated with only elevated MS-AFP, LBW associated with elevated MS-AFP, elevated MS-hCG, and lowered uE3, PIH associated with only elevated MS-AFP, PPROM only elevated MS-AFP, prematurity only elevated MS-AFP, and placenta previa over 35 years, elevated MS-hCG.(P <0.05). However, abruptio placenta was not significantly associated with predictor markers.(P >0.05) CONCLUSIONS: Some predictors such as over 35 year, elevated hCG, lowered uE3, a false positive screen for Down syndrome were significantly associated with adverse pregnancy outcome. Also in multivariate analysis, we identified especially elevated AFP to be the most reliable predictor for adverse pregnancy outcome.
alpha-Fetoproteins ; Biomarkers ; Chorion ; Down Syndrome ; Estriol ; Female ; Fetus ; Humans ; Logistic Models ; Mothers ; Multivariate Analysis* ; Parturition ; Placenta ; Placenta Previa ; Pregnancy ; Pregnancy Outcome* ; Pregnancy Trimester, Second ; Pregnancy* ; Radioimmunoassay ; Rupture

OBJECTIVE: This study was designed to detect genes specifically expressed in severe preeclamptic placentas. METHODS: Placenta tissues were collected immediately after delivery from 5 preeclamptic patients and 5 normal pregnant women. Total RNAs of each placenta were extracted and hybridized for a cDNA microarray. Of the microarray data, four up-regulated genes (DSCR4, GPA, PCDHGB1, Hemogen) and four down-regulated genes (IL1R2, MGST1, GAS1 GREB1) were selected and reverse transcriptase-polymerase chain reaction was used to confirm the results of cDNA microarray. RESULTS: The expression fold for each up-regulated gene was 2.2 times for DSCR4, 2.7 times for PCDHGB1, 3.5 times for Hemogen, 5.2 times for GPA on the cDNA microarray. The expression fold for each down-regulated gene was 3.3 times for IL1R2, 4.2 times for MGST1, 4.9 times for GAS1 and 2.3 times for GREB1 on the cDNA microarray. The expression fold for each up- regulated gene was 5.21 times for DSCR4, 3.01 times for PCDHGB1, and 4,53 times for Hemogen and 2.2 times for GPA on RT-PCR. The expression fold for each down-regulated gene was 2.7 times for IL1R2, 2.22 times for MGST1, 2.53 times for GAS1 and 1.83 times for GREB1 on the RT-PCR. CONCLUSION: DSCR4, PCDHGB1, Hemogen and GPA as the up-regulated genes and IL1R2, MGST1, GAS1 and GREB1 as the down-regulated genes, which were found and selected by the cDNA microarray, might be considered to be novel biomarkers for preeclampsia.
Biomarkers ; Chimera ; Female ; Gene Expression ; Humans ; Microarray Analysis ; Oligonucleotide Array Sequence Analysis ; Placenta ; Pre-Eclampsia ; Pregnant Women ; RNA

OBJECTIVETo establish the methods of Polychlorinated Dibenzo-p-Dioxins and Dibenzofurans (PCDD/Fs), polychlorinated biphenyls (PCBs) and polybrominated diphenyl ethers (PBDEs) compounds determination by isotope dilution HRGC/HRMS simultaneously in human placenta tissue from mothers, and assess the human exposure risk to dioxins and PBDEs in study.

METHODSConcentrations of 17 PCDD/Fs and 12 dioxin-like PCBs as well as 7 PBDEs were measured in human placenta tissue samples by isotope dilution HRGC/HRMS. SigmaTEQ (PCDD + PCDFs + PCBs) concentration using WHO-TEF factor and PBDEs concentration was calculated respectively. Risk assessment of mother exposure to dioxins and PBDEs was evaluated.

RESULTSMedian of SigmaTEQ (PCDD + PCDFs + PCBs) concentration for six samples was 18.15 WHO-TEQ pg/g lipid, ranging from 5.14 - 67.01 WHO-TEQ pg/g lipid. Although the median of SigmaTEQ (PCDD + PCDFs + PCBs) was lower than that of human blood of EU and Japan, and close to that of Korea and Taiwan non-exposure as reported in the literatures, the highest SigmaTEQ (PCDD + PCDFs + PCBs) concentration of placenta sample exceeded the value of high dioxins exposure area subjects in Taiwan. The dominant contributor congener for WHO-TEQ were 2, 3, 4, 7, 8-PeCDF, 1, 2, 3, 7, 8-PeCDD, PCB126, totally accounted for 65 percent of SigmaWHO-TEQ. Median and average of PBDE concentration for six samples were 2.73 ng/g lipid and 7.17 ng/g lipid, respectively, ranging from 0.95 - 25.99 ng/g lipid. BDE47 was the dominant contributor congener for the total concentration, accounted for 35 percent.

CONCLUSIONThe methods of PCDD/Fs, PCBs and PBDEs compounds determined by isotope dilution HRGC/HRMS simultaneously in human placenta tissue from mothers were established successfully, and the human exposure risk to PCDD/Fs, PCBs and PBDEs should be surveyed for the donor with the highest SigmaTEQ (PCDD + PCDFs + PCBs) and PBDEs concentration of placenta sample in the future.

Benzofurans ; analysis ; Female ; Halogenated Diphenyl Ethers ; analysis ; Humans ; Maternal Exposure ; Placenta ; chemistry ; Polychlorinated Biphenyls ; analysis ; Polychlorinated Dibenzodioxins ; analogs & derivatives ; analysis ; Pregnancy