Expression of Survivin in Early Villus and Decidua and Its Implication

Summary: 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 μg 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.

DETECTION OF STRAND BREAKS OF DNA IN HUMAN EARLY CHORIONIC VILLUS CELLS INDUCED BY DIAGNOSTIC ULTRASOUND USING ^(32)P-LABELED ALU HYBRIDIZATION

Objective To explore if strand breaks of DNA in human early chorionic villus cells in uterus were induced by diagnostic ultrasound and to evaluate the method used for detection of single-stranded breaks and double-stranded breaks in human DNA. Methods 60 normal pregnant women aged 20-30, who underwent artificial abortion during 6-8 weeks of gestation, were randomly divided into 2 experimental groups: All 30 cases were exposed to diagnostic ultrasound in uterus for 10 minutes, and 24 hours later chorionic villi were extracted; the other 30 cases were taken as the control group. Single-stranded DNA and double-stranded DNA in villus cells in all cases were isolated by the alkaline unwinding combined with hydroxylapatite chromatography, and were quantitatively detected using 32 P-labeled Alu probe for dot-blotting hybridization. Results There was no significant difference in quantity and percentage in single-stranded DNA and double-stranded DNA between 2 groups (P>0.05). 32 P-Alu probe could only hybridize with human DNA, and could detect DNA isolated from as few as 2.5×10 3 chorionic villus cells and 0.45ng DNA in human leukocytes. Conclusion The results suggested that there were no DNA strand damages in human chorionic villus cells when the uterus was exposed to diagnostic ultrasound for 10 minutes. The method,^(32)P-Alu probe for dot-blotting hybridization, was even more specific, sensitive and accurate than conventional approaches.

Relationship of Abortion and the Expression of Indoleamine 2,3- dioxygenase (IDO) in Villus and Syncytiotrophoblasts

Objective To study the relationship of abortion and the expression of indoleamine 2, 3- dioxygenase （IDO） in villus and syncytiotrophoblast in vitro. Methods RT-PCR was applied to analyze the mRNA transcription of lDO in villus of normal pregnancy and inevitable abortion and JAR cells as well. Immunohistochemistry was applied to analyze the expression of IDO protein in villus. Western blot was applied to determinate the expression of IDO protein on cultured syncytiotrophoblast. Highperformance liquid chromatography was applied to determinate whether there was kynurenine in cell culture medium of syncytiotrophoblast. Results The expression of IDO mRNA and protein in villus of inevitable abortion was lower than that of normal pregnancy; IDO mRNA did not express in JAR cells. IDO protein expressed on cultured syncytiotrophoblast, and there was kynurenine in cell culture medium of syncytiotrophoblast. Conclusion Appropriate expression of IDO in villus is necessary.for maintenance of normal pregnancy and an active IDO protein expresses in syncytiotrophoblast.

Outcome of 1355 consecutive transabdominal chorionic villus samplings in 1351 patients

Background The true risk of choronic villus sampling （CVS） is poorly defined. The objective of this study was to review the clinical outcome of transabdominal CVS performed in a university teaching unit, with an emphasis on the complication rate.Methods A comprehensive audit database was maintained for 1351 pregnant women, including 17 sets ot twin pregnancies, who had a CVS. Details and outcome of all CVSs made in the unit between May 1996 and May 2004 were reviewed. All CVSs were performed by one of 5 operators using the identical techniques.Results All procedures were performed transabdominally. A total of 1355 CVSs were performed because there were 4 dichorionic twin pregnancies which required 2 punctures. The mean gestation at CVS was （ 11.8 ＋ 0. 7 ） weeks, and 97.3% of the procedures were performed between 11 and 13 completed weeks. The majority （96. 2% ） required only 1 puncture to achieve correct needle placement. The procedure failed to obtain an adequate sample in 4 subjects （0.30%）. A total of 1351 chromosomal studies were requested and there was 1 case （0. 07% ） of culture failure. The results of chromosomal studies were available within 14 days in 36. 7% of the cases and within 21 days in 94. 0%. Overall, 77 chromosomal abnormalities （5.7%） and 5 cases of thalassemia major were detected. Pregnancy outcome was unknown in only 13 singleton subjects （0.96%）. In the remaining 1355 fetuses, there were 76 pregnancy terminations （5.56%）, 10 fetal losses with obvious obstetric causes （0. 73% ）, and 21 potentially procedure-related fetal losses （ 1.54% ）. In the last group, the majority had one or more co-existing obstetric complications. The background fetal loss rate for pregnancies at similar gestational age in the unit was about 0. 8%. Therefore, the procedure-related fetal loss rate was estimated to be at the maximum of 0. 74%.Conclusions In experienced hands, first trimester transabdominal CVS is an accurate and safe invasive prenatal diagnostic procedure. It should be one of the treatment options available to pregnant women who require prenatal genetic diagnosis.

A mathematical model for absorption of intestinal villus

Physiologists have done many researches on the mechanism of the intestinal villus absorption. Since the 1970s, Haljamae and Jodal presented serial experimental reports. They believed that there is a countercurrent mechanism in the intestinal villus, so the osmolality in the tip is 2—3 times greater than that in the base. In a conclusive article, Jodal believed that, in accordance with the anatomic considerations, the blood flows from the base to the tip of the intestinal villus in the central arterial vessel, and then descends from the tip to the base in the peripheral capillaries and veins, so that there are opposite blood flowing directions in the intestinal villus which makes the countercurrent mechanism possible. Second, the active pumping of NaCl and passive absorption of water

Effect of dietary vanadium on small intestinal morphology in broilers

The objective of this study was to determine the effects of dietary vanadium on small intestinal morphology of broilers by the methods of light microscopy (LM) and transmission electron microscopy (TEM). A total of 420 one-day-old avian broilers were divided into six groups (seven replicates in each group and ten broilers in each replicate) and fed on a control diet or the same diet supplemented with 5, 15, 30, 45 and 60 mg/kg vanadium in the form of ammonium metavanadate for 42 days. In comparison with those in the control group, the intestinal villus heights were decreased (P < 0.05 or P < 0.01) in the 30, 45 and 60 mg/kg groups, and crypt depths and villus height/crypt depth ratio were decreased in the 45 and 60 mg/kg groups. Ultrastructurally, the microvilli were apparently sparse and short, and the numbers of lysosomes were increased in abovementioned three intestines in the 45 and 60 mg/kg groups at 42 days of age. In conclusion, dietary vanadium in excess of 30 mg/kg could alter the villus height, crypt depth, villus height/crypt depth ratio and ultrastructure, which might impact the development of small intestines in broilers.

Acute effects of rotavirus and malnutrition on intestinal barrier function in neonatal piglets

AIM: To investigate the effect of protein-energy malnutrition on intestinal barrier function during rotavirus enteritis in a piglet model.METHODS: Newborn piglets were allotted at day 4 of age to the following treatments:(1) full-strength formula(FSF)/noninfected;(2) FSF/rotavirus infected;(3) half-strength formula(HSF)/noninfected;or(4) HSF/rotavirus infected.After one day of adjustment to the feeding rates,pigs were infected with rotavirus and acute effects on growth and diarrhea were monitored for 3 d and jejunal samples were collected for Ussingchamber analyses.RESULTS: Piglets that were malnourished or infected had lower body weights on days 2 and 3 post-infection(P < 0.05).Three days post-infection,marked diarrhea and weight loss were accompanied by sharp reductions in villus height(59%) and lactase activity(91%) and increased crypt depth(21%) in infected compared with non-infected pigs(P < 0.05).Malnutrition also increased crypt depth(21%) compared to full-fed piglets.Villus:crypt ratio was reduced(67%) with viral infection.There was a trend for reduction in transepithelial electrical resistance with rotavirus infection and malnutrition(P = 0.1).3H-mannitol flux was significantly increased(50%;P < 0.001) in rotavirus-infected piglets compared to non-infected piglets,but there was no effect of nutritional status.Furthermore,rotavirus infection reduced localization of the tight junction protein,occludin,in the cell membrane and increased localization in the cytosol.CONCLUSION: Overall,malnutrition had no additive effects to rotavirus infection on intestinal barrier function at day 3 post-infection in a neonatal piglet model.

Dyspepsia and celiac disease: Prevalence, diagnostic tools and therapy

The prevalence of dyspepsia is up to 40% in population-based study. Functional dyspepsia is an exclusion diagnosis and it is classified as a chronic abdominal pain-related functional disorder, characterized by the presence of persistent or recurrent pain or discomfort centered in the upper abdomen, neither relief by defecation, nor association with the onset of a change in stool frequency or form. Celiac disease(CD) is a common autoimmune enteropathy, with a prevalence around 1% in the general population. Its diagnosis includes a serological screening and an upper gastrointestinal endoscopy with multiple biopsies. Gluten-free diet is the only effective treatment. CD diagnosis is often delayed in asymptomatic patients or in individuals with less clinical gastrointestinal symptoms. Several studies performed coeliac disease screening in patients with symptoms suggestive of dyspepsia, showing a biopsy-proved prevalence that ranged from 0.5% to 2%. The typical endoscopic markers of villous atrophy are not sufficiently sensitive, so some endoscopic techniques, such as "water immersion" and confocal endomicroscopy were proposed to improve the diagnosticsensitivity and target biopsies. A recent meta-analysis estimated that the prevalence of CD was higher in patients with dyspepsia, but not in a statistically significant way. However this assumption should be confirmed further larger studies.

Biodegrading Wheat Bran with Agaricus blazei and Its Effects on Intestinal Development Identified with Mice

Agaricus blazei Murill is a well known edible/medicinal mushroom used for immune-nutrient food therapies.The biodegradation of different substrates with this fungus may result in different metabolites and degraded compounds,which may enrich the function of the food therapies.In this study,soluble compounds from the culture of A.blazei grown in liquid media with whole wheat bran and its water filtrate,respectively,were compared.Total soluble sugar,arabinoxylan,protein,and amino acids were significantly higher in the fungal culture resulted from the medium with whole wheat bran （43.54,1.56,0.59,and 2.19 mg mL^-1,respectively） than that from the medium with the bran filtrate （17.28,0.37,0.13,and 1.13 mg mL^-1,respectively）.The biodegraded wheat bran with cultured mycelia was fed to Mus musculus Linnaeus as supplementation or as dietary fiber exclusive ingredient.As non-specific food therapies,feeding effects on mice intestinal development were indicated indirectly in growth performance,intestinal absorption and serum parameters.Compared to feeding uncultured wheat bran,feeding mice with culture mixes of A.blazei resulted in remarkably increase in villus height and villus height/crypt depth ratio,which were increased by 25.4 and 31.0%,respectively,when applied as supplementation,and by 44.3 and 43.4%,respectively,when applied as dietary fiber.These increases are concomitant with the higher level of D-xylose in blood serum about 16.9 and 29.2% as supplementation and dietary fiber,respectively.The results implying that culturing A.blazei with whole wheat bran enhanced extracellular metabolism of the fungus and extensive degradation of wheat bran insoluble fibrous compounds.Furthermore,feeding the culture mix including metabolites and degraded wheat bran improved intestinal villus development,proving the nutritional benefits of the A.blazei mycelial cultures.

Expression of TET and 5-HmC in Trophoblast Villi of Women with Normal Pregnancy and with Early Pregnancy Loss

Increasing evidence suggests that epigenetic dysfunction may influence the stability of normal pregnancy. The ten-eleven translocation （TET） family and 5-hydroxymethylcytosine （5-hmC） were found to be linked with epigenetic reprogramming. The present study aimed to examine the expression of the TET family and 5-hmC in the villi of human embryos and compared their expression between normal pregnancy and early pregnancy loss （EPL）. Embryonic villi were collected from normal pregnant women （control） experiencing medical abortion and from EPL patients at gestation ages of 6, 7 and 8 weeks. The mRNAs of TET family were analysed using quantitative polymerase chain reaction （qPCR）, and TET proteins using Western blotting and immunohistochemical analysis. The MethylFlashTM Kit was used to quantify the absolute amount of 5-methylcytosine （5-mC） and 5-hmC. Our results showed that the expression of the TETs and 5-hmC in the normal villus decreased with increasing gestational age. Immunohistochemistry revealed that the TET proteins were expressed in the cytoplasm of trophoblasts and their expression was the highest in the 6-week tissue samples, which was consistent with the qPCR and Western blot results. The expression of TET1, TET2, and TET3 was lower in the villi in EPL group than in normal pregnancy group （P〈0.05 for all）. It was concluded that the TET family and 5-hmC are critical in epigenetic reprogramming of human embryo. The findings also suggest that a deficiency of TETs in the villus might be associated with human EPL.

Effects of Oldenlandia diffusa Polysaccharides on the Structures of Chick Small Intestine Tissues

In order to explore the application effect of Oldenlandia polysaccharides(OPS)in the production of AA broilers,2007-day-old AA broilers were randomly divided into 4 groups,namely groups Ⅰ,Ⅱ,Ⅲ,and Ⅳ,each including 50 chicks.The control group(group Ⅰ)was fed with basal diet;and the 3 experimental groups were fed based on the basal diet with the addition of OPS at 0.4 g/kg(group Ⅱ),0.8 g/kg(group Ⅲ),and 1.2 g/kg(group Ⅳ)feed for 1 week,respectively,and then fed the basal diet for 4 weeks.At the ages of 14,28,and 42 d,10 chickens were put into death in the control group and the experimental groups,respectively,and the duodenum and jejunum tissue samples were collected for the analysis of the effects of OPS on the structures of chick intestines.The results showed that at the age of 14 d,the villus lengths of duodenum and jejunum in groups Ⅱ,Ⅲ,and Ⅳ were significantly greater than those in group Ⅰ(P<0.05),and the ratios of duodenal or jejunal villus length to crypt depth in groups Ⅱ and Ⅲ were significantly greater than that in group Ⅰ(P<0.05);at the age of 28 d,the villus length of duodenum in group Ⅱ was significantly greater than that in group Ⅰ(P<0.05);and at the age of 42 d,the ratios of jejunual villus length to crypt depth in groups Ⅱ,Ⅲ and Ⅳ were significantly larger than those of group I(P<0.05).It showed that OPS could promote the small intestine development of AA broilers aged 14 to 42 d.

Invasive Procedures for Prenatal Diagnosis in Salmaniya Medical Complex in Bahrain: A Retrospective Cross-Sectional Descriptive Study

Background: Prenatal diagnosis is the process of evaluating the presence of disease or potential disease in the fetus, this enables families to be better prepared before the birth of the baby. There are non-invasive prenatal diagnosis procedures and invasive prenatal diagnosis procedures. The invasive prenatal diagnosis procedures are CVS (chorionic villus sampling) and amniocentesis. The American College of Obstetricians and Gynecologists states that invasive diagnostic testing should be available to all women, regardless of age or risk. Objective: To determine the indications, outcome and results of diagnostic invasive prenatal procedures. Study setting: The obstetrics and Gynecology Department in Salmaniya Medical Complex in Kingdom of Bahrain. Study design: Retrospective descriptive study. Study subjects and Methods: This retrospective descriptive study was conducted on 175 pregnant women who underwent invasive prenatal procedures (CVS and amniocentesis) between January 2013 and December 2018 at SMC in Kingdom of Bahrain. All medical records of the participants were reviewed and entered the study. According to the implemented procedures, medical records were categorized into two chorionic villus sampling (CVS) and amniocentesis groups. The study subject will include indications of the procedures which are advanced maternal age, hematological disorders, genetic disorders, metabolic disorders, abnormal structural findings in fetal ultrasound and previous child with aneuploidy. In addition, the study will address the complications, outcome and results of procedures. Results: About half of our indications of the procedures were due to hematological disorders (47.6%) followed by abnormal structural findings in fetal ultrasound (30.1%) then genetic disorders (15.7%), metabolic disorders (4.8%) and advanced maternal age (1.8%). Regarding complications of the procedure;threatened miscarriage or loss of pregnancy within 3 weeks was (2.3%), amniotic fluid leakage (0.7%), abdominal cramps (0.7%) and Insufficient or contaminated sample (6.2%). Regarding outcome of the pregnancy, our results showed that the loss of pregnancy was (4.8%), intrauterine fetal death or still birth was (13.9%), live birth was (63.9%), preterm delivery was (7.8%), preterm premature rupture of membrane (PPROM) was (1.8%), limbs reduction was (0.0%). Termination of pregnancy outside the country was (7.8%) of chorionic villus sampling and amniocentesis. Conclusion: CVS and amniocentesis are useful outpatient procedures to detect diagnosis or to assess whether a patient is at increased risk of having an affected fetus and that will minimize the psychological impact on the patient and to provide a proper antenatal care to the pregnant women by her obstetrician and follow up to the baby by pediatrician. In this study it was observed that most of the patients who underwent the procedure were couples either carrier or affected to sickle cell disease or Beta thalassemia.

Prenatal cytogenetic diagnosis study of 2782 cases of high-risk pregnant women

Background Prenatal diagnoses are extremely advantageous for pregnant women with high-risk indicators and can help prevent the birth of malformed infants. However, no large-scale statistical study analyzing the correlation between fetal chromosome disorders and abnormal indicators during pregnancy has been done in China. The objectives of this study were to diagnose and analyze fetal chromosome abnormalities, determine the feasibility of the various prenatal test methods and establish diagnostic guidelines for the early, middle, and late trimesters. Methods From January 2004 to May 2009, 2782 pregnant women at high-risk underwent prenatal diagnoses. Categorized data expressed as either actual counts or percentages were analyzed by the chi-square or Fisher＇s exact test. Chorionic villus sampling was performed in the early-trimester （10-12 weeks of gestation）, amniocentesis in mid-trimester （16-28 weeks of gestation）, and umbilical cord blood collection in mid- or late-trimester （16-37 weeks of gestation）. In 51 cases either autopsy samples from intrauterine fetal deaths or placental tissues from aborted fetuses were tested. Results Chromosomal abnormalities were observed in 3.99% （111/2782） of the samples. Overall, the success rate of cytogenetic analysis for high-risk pregnancy groups was 98.17% （2731/2782）. It was significantly less successful when used to analyze data from the chorionic villus sampling compared with that from amniocentesis and umbilical cord blood （P=-0.000）. Abnormal chromosome carriers had the highest percentage of abnormal chromosomes （67.86%） when compared with chromosomal abnormalities in patients with ultra-sonographic ＂soft markers＂ （11.81%）, advanced maternal age （4.51%） and those who had positive serum screening results （P=-0.000）. Conclusions Invasive prenatal diagnostic techniques are feasible tools for confirming fetal chromosomal abnormalities. Abnormal chromosomes detected in one of the parents carrying abnormal chromosome, ultrasound soft markers, advanced maternal age or positive serum screening results were associated with a higher frequency of fetal genetic diseases.

Expression and Influence of Galectin-3 on Missed Abortion

Objective To explore the influence of galectin-3 on missed abortion. Methods Forty cases of normal intrauterine early pregnancy were randomly divided into 2 groups： surgical abortion group （group A, n=20） and medical abortion group （group B, n =20）. The third group was missed abortion group （group C, n =20） with the gestational age less than 13 weeks. Serum was isolated from the blood samples, collected and used for ELISA quantification of galectin-3. Villus and decidua tissues were collected from the abortus for immunohistochemical examination and real-time fluorescence relative quantitative PCR. Results The level of galectin-3 in the serum was the lowest in missed abortion group （P〈0.05）. Immunohistochemistry showed that galectin-3 expression in villus of missed abortion group was significantly lower than that of surgical abortion group （P〈0. 01）. Real-time fluorescence relative quantitative PCR showed that galectin-3 mRNA relative expression in villus of missed abortion group （2^-△△Ct=0. 04± 0. 01） was significantly lower than that of surgical abortion group （2^-△△Ct=1.00 ± 0.00）. Galectin-3 mRNA relative expression in deciduas of medical abortion group （2-zact=o. 08 ± 0.02） was s!gnificantly lower than that of surgical abortion group （2^-△△Ct=1.00 ± 0.00） （P〈0. 01）. Conclusion Galectin-3 is related to the development of villus and decidua during early pregnancy. The decreased expression of galectin-3 may promote the occurrence of missed abortion.

Prenatal Testing or Screening?

Over the past 50 years,the scope and extent of prenatal diagnosis and screening for genetic disorders have improved geometrically.There has been a pendulum like swing from testing to screening back and forth as new technologies emerge.The concurrent developments of cell free fetal DNA analysis of maternal blood has dramatically changed patient’s choices towards screening.However,with the use of array comparative genomic hybridization of fetal DNA that requires diagnostic procedures(Chorionic villus sampling and amniocentesis),much more extensive diagnosis can be obtained.Until noninvasive methods can replicate what can be done with diagnostic procedures there still will be a"price to be paid"for opting for the non-invasive methods.