Bile Acid Effects on Placental Damage in Intrahepatic Cholestasis of Pregnancy

Aims: The abnormal increase of bile acid is found in intrahepatic cholestasis of pregnancy (ICP). It also can be observed the damage of placental tissue in ICP. The aim of this study was to find the associations of the bile acid in umbilical vein and the damage of placental tissue. Methods: Thirty women diagnosed with ICP and fifty normal pregnant women between September 2015 and September 2017 at Nanshan District Maternity & Child Healthcare Hospital of Shenzhen were included in this study. The glycocholic acid (GA), total bile acids (TBA), total bilirubin (TB), direct bilirubin (DB) and albumin level in umbilical vein were measured by cycle enzyme method in ICP and control group. The placental damage was analyzed by morphologic study using hematoxylin dyes in two groups. The correlation between the level of the bile acid in the umbilical vein and the damage of the placenta was assessed using SPSS software. Results: The GA, TBA, TB, DB and albumin level in umbilical vein were significantly higher in ICP than those of pregnant women, respectively. The placental villis were expanded and the structure was destroyed in ICP. The vessel was damaged and the cell trophoblast hyperplasia in ICP. It also can be seen that there was obvious nodules and a typical fibrous necrotic substance in ICP but not in control group. There is a positive correlation between the level of the TBA in the umbilical vein and the damage of the placenta in ICP. Conclusion: The TBAs were significantly higher in umbilical vein and were related to the placental damage in ICP.

Effects of creep feeding and supplemental glutamine or glutamine plus glutamate (Aminogut) on pre- and post-weaning growth performance and intestinal health of piglets

Background： Creep feeding is used to stimulate piglet post-weaning feed consumption.L-Glutamine（GLN） is an important source of fuel for intestinal epithelial cells.The objective of this study was to determine the impact of creep feeding and adding GLN or AminoGut（AG;containing glutamine ＋ glutamate） to pre-and post-weaning diets on pig performance and intestinal health.Litters（N = 120） were allotted to four treatments during 14–21 d of lactation： 1） No creep feed（NC,n = 45）;2） creep fed control diet（CFCD,n = 45）;3） creep fed 1% GLN（CFGLN,n = 15）;4） creep fed.88% AG（CFAG,n = 15）.After weaning,the NC and CFCD groups were sub-divided into three groups（n = 15 each）,receiving either a control nursery diet（NC-CD,CFCD-CD） or a diet supplemented with either GLN（NC-GLN,CFCD-GLN） or with AG（NC-AG,CFCD-AG）.Litters that were creep fed with diets containing GLN or AG also were supplemented with those amino acids in the nursery diets（CFGLN-GLN,CFAG-AG）.Glutamine was added at 1% in all three post-weaning diet phases and AG was added at.88% in phase 1 and 2 and at.66% in phase 3.Results： Feed conversion（feed/gain） showed means among treatment means close to significance（P = 0.056） and Tukey＇s test for pairwise mean comparisons showed that Pigs in the CFGLN-GLN group had the best feed conversion（feed/gain） in the first three-week period post-weaning,exceeding（P = 0.044） controls（CFCD-CD） by 34%.The NC-AG group had（P = 0.02） the greatest feed intake in the last three week of the study,exceeding controls（CFCD-CD） by 12%.CFGLN-GLN,CFCD-GLN and sow reared（SR） pigs had the greatest（P = 0.049） villi height exceeding the CFCD-AG group by 18%,20% and 19% respectively.The CFAG-AG group had the deepest（P = 0.001） crypts among all treatments.CFGLN-GLN,CFCD-GLN and SR groups had the greatest（P = 0.001） number of cells proliferating（PCNA） exceeding those in the NC-CD group by 43%,54% and 63% respectively.Sow reared pigs showed the greatest（P = 0.001） intestinal absorption capacity for xylose and mannitol.Conclusion： Supplementation of creep feed and nursery diets with GLN and/or AminoGut in the first three week improved feed conversion possibly due to improved intestinal health.

CXCL12/CXCR4 Expression in Trophoblasts Takes Part in Materno-fetal Immune Tolerance and Vascular Remodeling

Summary： In this study, we investigated the expression of CXCL12 （SDF-1）/CXCR4 in trophoblasts and the role they play in the gestation. Immunochemistry was used to detect the expression of CXCR4 and CXCLI 2 in human villi and placenta. Highly purified extra-viUous trophoblasts （EVTs） ere detected for CXCR4 and CXCL12 in vitro by immunocytochemistry. The chemotaxis of CXCL12 was tested in transweU and the chemotactic activity was quantitatively examined. It was suggested that both CXCR4 and CXCL12 were expressed in trophoblasts and were decreased with the gestation time P〈0.05）. In a certain coverage, CXCL12 exhibited chemotactic activity which was positively correlated with its concentration [（r）=0.68, P〈0.01], the maximum chemotactic index （CI） was 1.62±0.12. Our results suggest that interaction between CXCR4 and CXCL12 is involved in materno-fetal immunological tolerance in all three trimesters of gestation and contributes to the invasion of EVTs during pregnancy.

Clonal isolation of endothelial colony-forming cells from early gestation chorionic villi of human placenta for fetal tissue regeneration

BACKGROUND Endothelial colony-forming cells(ECFCs)have been implicated in the process of vascularization,which includes vasculogenesis and angiogenesis.Vasculogenesis is a de novo formation of blood vessels,and is an essential physiological process that occurs during embryonic development and tissue regeneration.Angiogenesis is the growth of new capillaries from pre-existing blood vessels,which is observed both prenatally and postnatally.The placenta is an organ composed of a variety of fetal-derived cells,including ECFCs,and therefore has significant potential as a source of fetal ECFCs for tissue engineering.AIM To investigate the possibility of isolating clonal ECFCs from human early gestation chorionic villi(CV-ECFCs)of the placenta,and assess their potential for tissue engineering.METHODS The early gestation chorionic villus tissue was dissociated by enzyme digestion.Cells expressing CD31 were selected using magnetic-activated cell sorting,and plated in endothelial-specific growth medium.After 2-3 wks in culture,colonies displaying cobblestone-like morphology were manually picked using cloning cylinders.We characterized CV-ECFCs by flow cytometry,immunophenotyping,tube formation assay,and Dil-Ac-LDL uptake assay.Viral transduction of CVECFCs was performed using a Luciferase/tdTomato-containing lentiviral vector,and transduction efficiency was tested by fluorescent microscopy and flow cytometry.Compatibility of CV-ECFCs with a delivery vehicle was determined using an FDA approved,small intestinal submucosa extracellular matrix scaffold.RESULTS After four passages in 6-8 wks of culture,we obtained a total number of 1.8×107 CV-ECFCs using 100 mg of early gestational chorionic villus tissue.Immunophenotypic analyses by flow cytometry demonstrated that CV-ECFCs highly expressed the endothelial markers CD31,CD144,CD146,CD105,CD309,only partially expressed CD34,and did not express CD45 and CD90.CV-ECFCs were capable of acetylated low-density lipoprotein uptake and tube formation,similar to cord blood-derived ECFCs(CB-ECFCs).CV-ECFCs can be transduced with a Luciferase/tdTomato-containing lentiviral vector at a transduction efficiency of 85.1%.Seeding CV-ECFCs on a small intestinal submucosa extracellular matrix scaffold confirmed that CV-ECFCs were compatible with the biomaterial scaffold.CONCLUSION In summary,we established a magnetic sorting-assisted clonal isolation approach to derive CV-ECFCs.A substantial number of CV-ECFCs can be obtained within a short time frame,representing a promising novel source of ECFCs for fetal treatments.

Effect of Mifepristone on Transforming Growth Factor -βand its Receptor Transcription in Decidua, Villi and Serum Tumor Necrosis Factor -α Level in Early Human Gestation

Objective To investigate the role of mifepristone in regulating cytokines of materno-fetal interface and serum of human early gestationMethods Thirty-five women with early pregnancy received mifepristone 50 mg orallyon study d 1 and d 2, respectively, followed by undergoing artificial abortion to getdecidua and villi on study d 3. Twenty-five women with early pregnancy withoutmifepristone administration as control also underwent artificial abortion to get de-cidua and villi. The expressions of TGF-β1 and TGF-β1 receptor mRNA in the earlydecidua and villi were assessed by using RT-PCR . The concentrations of serum TNF-α were measured by radioimmunoassay.Results The decidual expressions of TGF- β1 mRNA and TGF-β1 receptor mRNA in thetreated group were significantly lower than those of the control (P<0.05), while thevillus expressions of TGF-β1 and TGF-β1 receptor mRNA in the treated group were notsignificantly different from those of the control (P>0.05). The serum TNF-β1 levelselevated significantly after mifepristone treatment.Conclusion The antigestational effect of mifepristone might act through suppressingthe transcription of TGF-β1 and TGF-β1 receptor in the decidua and increasing theserum TNF-α level, which interfered in the materno-fetal interface Th2 bias.

A Srudy on Ultrastructures of Normal 5-8 Week Human Placental Villi

Fourteen normal human placental villi at a fetal age of 5-8 weeks were examined withelectron microscope. The results were vide infra.(1)The cytotrophoblasts were characterized by largenucleus with less heterochromatin and prominent nucleolus with a number of free polyribosomes as well asscattered mitochondria in the cytoplasm.(2) There was a layer of dense microvilli on the surface of thesyncytiotrophoblastic cells.The size and shape of microvilli were variable. Usually fine globular expansions were present at the apical pole of microvilli and the cell membrane between the base of microvillihad invaginations into the cytoplasm forming small vesicles and tubules. Rough endoplasmic reticulum,free polyribosomes,mitochondria, lipid droplets and few myelinlike figures were also found in the cytoplasm.Part of cytoplasm penetrated into the interspaces of cytotrophoblastic cells and became in directcontact with the basement membrane.(3) Numerous desmosomes were seen between the cells of cytotrophoblast and syncytiotrophoblast.And (4) in the strom a of villi,mesenchymal cells were abundant inrough endoplasmic reticulum and well developed Golgi complex. Mesenchymat cells were markedlyfunctionally active. Ultrastructural characteristics of placental villin in connexion with their functionwere discussed.

The physiological characteristics of the basal microvilli microvessels in pancreatic cancers

Pancreatic cancer(PC)is a highly lethal tumor with controversial high glucose uptake and hypomicrovascularity.And,the hypomicrovasculature,which is considered to have poor perfusion,blocks the delivery of drugs to tumors.Previously,we described the preferential existence of a novel endothelial projection with trafficking vesicles in PCs,referring to basal microvilli.However,whether the basal microvilli microvessels have good perfusion or deliver nutrients to the tumor milieu is unknown.Here,we perfused multiple endothelial markers and nutrients to autochthonous PC-bearing mice to study the nutrient trafficking and perfusion status of the basal microvilli microvasculature.

Purification and Properties of Human Liver Glutathione Peroxidase

Fourteen normal human placental villi at a fetal age of 5-8 weeks were examined withelectron microscope. The results were vide infra.(1)The cytotrophoblasts were characterized by largenucleus with less heterochromatin and prominent nucleolus with a number of free polyribosomes as well asscattered mitochondria in the cytoplasm.(2) There was a layer of dense microvilli on the surface of thesyncytiotrophoblastic cells.The size and shape of microvilli were variable. Usually fine globular expansions were present at the apical pole of microvilli and the cell membrane between the base of microvillihad invaginations into the cytoplasm forming small vesicles and tubules. Rough endoplasmic reticulum,free polyribosomes,mitochondria, lipid droplets and few myelinlike figures were also found in the cytoplasm.Part of cytoplasm penetrated into the interspaces of cytotrophoblastic cells and became in directcontact with the basement membrane.(3) Numerous desmosomes were seen between the cells of cytotrophoblast and syncytiotrophoblast.And (4) in the strom a of villi,mesenchymal cells were abundant inrough endoplasmic reticulum and well developed Golgi complex. Mesenchymat cells were markedlyfunctionally active. Ultrastructural characteristics of placental villin in connexion with their functionwere discussed.

The contents and effect of angiotensin Ⅱ on hCG release in human placental villi during the first trimester of gestation

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High Expression of hsMAD2 in the Villi of Spontaneously Aborted Embryo with Chromosomal Abnormality

Objective: To investigate the changes of hsMAD2 protein and gene expression levels during chromosome segregation of human embryos. Method: The embryos of spontaneous abortion were collected in our hospital from 2009 to 2013, the chromosomal numbers of the embryonic villi were subsequently detected by fluorescence in situ hybridization (FISH). The patients were then divided into the normal and abnormal groups based on the chromosome number. The hsMAD2 protein and gene expression levels in the villi tissues of the two embryo groups were detected by western blotting and qRT-PCR, respectively. The hsMAD2 protein and gene levels in the embryonic villus tissue of the patient were detected. Results: From 2009 to 2013, we collected 50 embryos from spontaneous abortion patients. The chromosome abnormality and no abnormality were 36 cases (abnormal number of 28 cases (56.0%) and chimerism in 8 cases (16.0%)) and 14 cases (28.0%), respectively. The expression of hsmad2 protein and its gene in the villi of spontaneously aborted embryo with chromosomal abnormality in the abnormal group was significantly higher than that in those without chromosomal abnormalities (0.88 ± 0.20 vs 0.61 ± 0.19, P < 0.05), (23.46 ± 0.07 vs 18.35 ± 0.10, P < 0.05). Conclusion: Abnormal number of chromosomes is closely related to spontaneous abortion Linked, hsMAD2 factor has a card effect on the cell cycle, can block the mitotic process of cells, and play an important role in maintaining the normal separation and stability of chromosomes.

The Influences of Mifepristone, Norethisterone and Tamoxifen on the Glycosph

By using the Ladisch partitioning and microscale-analysis of HPTLC,the comparative quantitative and qualitative studies of gangliosides (Gg)and neutral glycosphingolipids(N-GSL)compositions from human chorionic villi tissues of normal early pregnant women and women treated with mifepristone, norethisterone(NET)and tamoxifen(TAM)were reported in this paper.The patterns of Gg and N-GSL in three treated groups were similar to those in normal pregnant group.The total values of Gg from the chorionic villi tissues reduced significantly in three treated groups(P<0.01).In all treated groups,the amounts of NeuNAC-Gal-Glc-cer(GM3),NeuNAC-NeuNAC-Gal-Glc-cer(GD3)and Gal-GalNAC-(NeuNAC)-Gal-Glc-cer(GM1)were decreased significantly compared with those in normal(P<0.01orP<0.05).In NET and TAM groups,Neu NAC-Gal-GalNAC-(NeuNAC-NeuNAC)-Gal-Glc-cer(GT1b) was markedly lower than that in normal(P<0.01).The total values of N-GSL extracted from the chorionic villi tissues were obviously higher in mifepristone and TAM groups than those in normal(P<0.01).The Gal-Glc-cer(LacCer)(CDH)and Gal-Gal-Glc-cer(Gal-LacCer)(CTH)were greatly increased in mifepristone group as compared with normal P<0.05).Paragloboside:Gal-GalNAC-Gal-Glc-cer(PG)in NETgroup was significantly higher than that in normal(P<0.01).

中西医结合治疗小儿肺炎126例

中西医结合治疗小儿肺炎１２６例李维茹，王霞乌盟达茂旗医院（０１２０００）１９９１年８月１０日收稿小儿肺炎是儿科一种常见病，多发病。死亡率较高，近几年我科应用中西医结合的疗法，取得了良好效果．现总结如下：临床资料从１９９２年１月至１９９４年阴，收治小儿...

中西医结合治疗小儿肺炎96例

中西医结合治疗小儿肺炎９６例赵永胜，韩志远，降永胜达茂旗医院中医科（０１２０００）１９９１年１０月２０日收稿自１９８９年１０月以来，笔者采用中西医结合方法，以中医辨证论治为主，结合西药对症治疗小儿肺炎９６例，疗效满意，现介绍如下：１临床资料９６例小儿...

Expression and molecular mechanism of PCNA,Caspase-3,IL-6 and Survivin proteins in chorionic villi and decidual tissue of early embryo damage

Objective:This study aims to explore its role in embryo damage and the relationship between them by testing the expression of PCNA,Caspase-3,IL-6 and Survivin protein in the chorionic villi and decidual tissue of patients with unexplained early embryo damage and normal early pregnancy at the same time voluntarily requested uterine aspiration abortion in order to clarify the pathogenesis of early embryo damage from the cellular and molecular perspectives,and provide theoretical basis for the diagnosis and treatment of embryo damage.Methods:From July 2018 to July 2019,30 patients with unexplained embryo damage were selected as embryo damage group,thirty normal early pregnancy patients with aspiration abortion were selected as the normal early pregnancy group.The decidual tissue and chorionic villi of the two groups were collected to observe the structural and pathological changes.Immunohistochemical method was used to detect the distribution of PCNA,Caspase-3,IL-6 and Survivin in the chorionic villi and decidual tissue,as well as the expression changes of the above four proteins.Results:Compared with the normal early pregnancy group,the chorionic villi in the early embryo damage group were obviously dysplasia,degenerative changes,structural disorders,homogeneous destruction,obvious reduction or even disappearance of trophoblast cells,inflammatory cell infiltration,more neutrophils and lymphocytes,interstitial edema and cell atrophy,accompanied by hemorrhage;The decidual tissue of the intima was not good,most of the cells were spindle-shaped,the structure was disordered,the stroma has edema,inflammatory cells can be seen,and hemorrhage existed.The results of immunohistochemistry showed that the expression of PCNA protein in chorionic villi and decidual tissue of early embryo damage group decreased significantly(P<0.05),while the expression of Caspase-3 protein increased significantly(P<0.05).The expression of IL-6 protein decreased significantly(P<0.05),and the expression of Survivin protein decreased significantly(P<0.05).Conclusion:During early pregnancy,due to the down-regulation of the expression of proliferating cell nuclear antigen PCNA and apoptosis inhibitor protein Survivin in chorionic and decidual tissues,the balance between proliferation and apoptosis is broken,which has an increase in Caspase-3 expression and a decrease in IL-6 expression.It may be that the balance of Th1/Th2 in chorionic and decidual tissues inclines to Th1,which leads to excessive apoptosis of chorionic and decidual tissues and the cessation of early embryo damage.

Histomorphometric study of role of lactoferrin in atrophy of the intestinal mucosa of rats

The purpose of this work was to study the effects of oral administration of lactoferrin (Lf) in rats subjected to atrophy of the small intestine induced by a diet based on soy protein concentrate as the main protein source. We used 24 male Wistar rats aged 40 days, kept in individual cages under appropriate conditions of temperature, light and humidity. The animals were divided into four groups (n = 6);1) group SL received soy-based food and, once a day, a supplement of 200mg/kg of Lf administered by gavage;2) group Si received the soy feed without supplement of Lf;3) group CL received a diet based on casein plus Lf;4) group Ci received the casein diet without supplement of Lf. At the end of fifteen days, a 10 mm segment of the initial portion of the small intestine was sectioned and subjected to morphometry of the intestinal crypts and villi and assessment of the number and size of myofibroblasts. Comparison between groups showed that the length of the villi was similar in groups Ci and CL and higher in CL than in SL;SL than in Si, in Ci than in SL, and in Ci than in Si to Ci. The crypt depth was similar in SL and CL, SL and Ci and Ci and CL and was higher in Si than in Ci and in Si than in SL. The number of myofibroblasts was higher in SL than in CL, in SL than in Si, in CL than in Ci, and in SL than in and Ci;between Ci and Si there was no difference. The area of myofibroblasts was similar between the groups SL and CL and Si and Ci and higher in SL than in Si, and in Cl than in and Ci, and in SL than in Ci. All statistical analysis assumed significance when p < 0.05. From these results, we conclude that lactoferrin increases the number and size of the pericrypt myofibro-blasts and stimulates rapidly the regeneration of atrophied villi.

FETAL SEX DETERMINATION IN THE FIRST TRIMESTER OF PREGNANCY USING A Y CHROMOSOME-SPECIFIC DNA PROBE

Technique of recombinant DNA has been used for prenatal diagnosis of many kinds of genetic disorders and was also applied to the determination of fetal sex. In China, prenatal diagnosis of α-and β-thalassemia was done by this technique, but sex determination of fetus is still performed, by using chromosome analysis. The

Immunohistochemical localization of dopamine and norepinephrine receptors in human early placental villi

Evidence so far accumulated suggests that the human early placenta can be considered as a complicated neuroendocrine organt. Our previous reports have demonstrated that the human early placenta presents not only a variety of neuropeptides but also neurotransmitters such as 5-hydroxytryptamine (5-HT), dopamine (DA) and norepinephrine (NE). It has been shown that these neurotransmitters in human placenta may play an important role in the process of blastocyst implantation and maintenance of normal pregnancy. As far as we are aware, no information concerning the DA and/or NE receptors localized in

Immunohistochemistry of Dopamine and Norepinephrine in Human Placental Villi

Most of the reports on human placental villi endocrinology focus on the study of neuropeptides, pituitary-or gonadal-like hormones, growth factors and cytokines, and only a few of them concern the function of biogenic amine. We first discovered that trophoblast of placental villi can produce 5-hydroxytryptamine (5-HT) and that its

ASAP3 regulates microvilli structure in parietal cells and presents intervention target for gastric acidity

Gastric acidity-associated disorders such as peptic ulcer and reflux diseases are widespread,and the reported resistance and side effects of currently used medicines suggest an urgent requirement for alternative therapeutic approaches.Here we demonstrate a critical role of ASAP3 in regulating the microvilli structure of parietal cells in vivo,and reveal the feasibility of controlling gastric acidity by targeting ASAP3.Conditional knockout of ASAP3 in mice caused elongation and stacking of microvilli in parietal cells,and substantially decreased gastric acid secretion.These were associated with active assembly of F-actin caused by a higher level of GTP-bound Arf6 GTPase.Consistently,a small molecular compound QS11 inhibited ASAP3 function and significantly reduced gastric acidity in vivo.Of note,the expression of ASAP3 was positively correlated with gastric acid secretion in 90 human cases,and high expression of ASAP3 was associated with reflux disease and peptic ulcer.These results reveal for the first time that ASAP3 regulates the microvilli structures in parietal cells.Our data also suggest ASAP3 as a feasible and drugable therapeutic target for gastric acidity-associated diseases.

Research on the Expression of HOXA10 in Placental Villi during the Onset of Labor

Objective To investigate the difference of the expression of HOXAIO gene in placental villi before and after the onset of labor. Methods Immunohistochemistry was used to detect the location of HOXAIO in placental villi. The expression of HOXAIO mRNA and protein were measured by RT-PCR and Western blotting. Results lmmunohistochemistry showed that HOXAIO was only weakly expressed in placental villi of the natural labor group. However, it was strongly expressed in placental Villi of the surgical labor group. RT-PCR and Western blotting showed that the expression of HOXAIO mRNA and protein were significantly lower in the natural labor group than in the surgical labor group （P〈0.05）. Conclusion The lower expression of HOXAIO during the onset of labor may be one of the major factors in initiating the normal labor. And to some extent, the lower expression of HOXAIO gene in the natural labor group may be related to ＂functional progesterone withdrawal＂.