ABSTRACT
Objective During pregnancy , exosomes can be released from the placenta into maternal circulation and play im-portant roles in normal pregnancy or placenta-related diseases .We aimed to establish a simple and efficient method for isolating and i-dentifying placental exosomes from maternal serum and lay a foundation for the studies of pregnancy -related diseases . Methods Using sucrose gradient centrifugation with 8% PEG6000 precipitation twice , we isolated and purified placenta-derived exosomes from normal maternal serum and detected their molecular markers CD 63 , CD81 and PLAP by Western blot , followed by silver staining anal-ysis of the protein profile of the exosome pellet .We identified the morphology of the placenta-derived exosomes by transmission electron microscopy ( TEM) and measured the size and distribution of the particles by dynamic light scattering ( DLS) . Results Silver stai-ning of the protein profiles of the exosomes after sucrose gradient centrifugation clearly revealed the bands of the protein molecules . Western blot showed the expressions of CD 63, CD81, and PLAP in the 21-34%density layer, which demonstrated the presence of serum placental exosomes mainly in the 1.09-1.16 g/mL density layer.TEM exhibited that the placenta-derived exosomes were round or oval cup-shaped, specifically expressing PLAP, and the particles were uniform in size, with a mean diameter of (41.79 ±11.94) nm. Conclusion A simple, fast, and efficient method was successfully established for isolating placenta-derived exosomes from ma-ternal serum, which provides a basis for studying the roles of placental exosomes in normal pregnancy and placenta -related diseases.
ABSTRACT
Objective The CYP19 gene product enzyme aromatase mediates the conversion of the androgen testosterone to es -tradiol.The aim of this study is to investigate whether the CYP19A1 gene single nucleotide polymorphism (SNP) is associated with the susceptibility to polycystic ovary syndrome (PCOS) and serum hormone levels. Methods We conducted a case-controlled study, which included 373 PCOS patients and 313 healthy controls.We genotyped SNP rs2899470 in the subjects using the polymerase chain re-action-restriction fragment length polymorphism ( PCR-RFLP) method and analyzed the frequencies of genotypes and alleles as well as the association of different genotypes with age , menarchal age, body mass index (BMI), and serum levels of hormones. Results The gen-otypic distributions of rs2899470 GG, TG, and TT in the PCOS women were 44.5%, 49.6%, and 5.9%, respectively, significantly dif-ferent from 39.3%, 48.6%, and 12.1%in the healthy controls (P=0.013).The frequency of the G allele was 69.3%in the former, remarkably higher than 63.6%in the latter (P=0.025).The rs2899470 genotypic frequencies were associated with the serum E 2/T lev-els in the PCOS patients. Conclusion SNP rs2899470 in the CYP19A1 gene is associated with the susceptibility to PCOS , and so is the genotype of rs2899470 with serum E2/T levels, which may be attributed mainly to the reduced activity of aromatase .
ABSTRACT
Objective Preoperative autologous blood donation ( PABD) may reduce the need for allogeneic blood , but it may also cause a short massive blood loss in pregnant women , and its fetal and maternal safety has to be adequately assessed .This study was to evaluate the feasibility and safety of PABD for pregnant women and their fetuses . Methods A prospective observational study was conducted among the women who met the inclusion criteria and gave birth in Nanjing Drum Tower Hospital between January and December 2013 .According to the clinical validation of risk stratification criteria for peripartum hemorrhage of California 2013 , the ca-ses were classified into a low-, a medium-, and a high-risk group.Data on blood donation procedures , obstetric outcomes, and blood transfusions were collected after delivery for analysis . Results Totally, 92 pregnant women accomplished 115 blood donations .The median volumes of the donated blood were 300, 300, and 400 mL in the low-, medium-, and high-risk groups, respectively ( P>0.001).There were no significant changes in HR , SBP and SpO2 during the blood donation procedures (P>0.05) except for the fall of diastolic blood pressure by an average of 3.4 mmHg (P0.05), which were similar to those in the cases who donated twice , with no significant differences before and after the donation (P>0.05). Homologous blood transfusion was performed for 5 cases (17.9%) in the high-risk group, with the volume of blood loss >2000 mL in all the cases.All the newborns survived without asphyxia and there was no perinatal death . Conclusion PABD can provide timely autologous whole blood donation for pregnant women .Under strict management , PABD is feasible and safe for pregnant patients who are at a high risk for massive blood loss during delivery or have a rare type of blood no readily available .