ABSTRACT
To develop Senecavirus A (SVA) virus-like particles (VLPs), a recombinant prokaryotic expression plasmid pET28a-SVA-VP031 was constructed to co-express SVA structural proteins VP0, VP3 and VP1, according to the genomic sequence of the field isolate CH-FJ-2017 after the recombinant proteins were expressed in E .coli system, and purified by Ni+ ion chromatographic method. The SVA VLPs self-assemble with a high yield in vitro buffer. A typical VLPs with an average diameter of 25-30 nm which is similar to native virions by using TEM detection. Animals immunized by SVA VLPs shown that the VLPs induced high titers neutralizing antibodies in Guinea pigs. This study indicated that the VLPs produced with co-expressing SVA structural proteins VP0, VP3 and VP1 in prokaryotic system is a promising candidate and laid an important foundation for the development of a novel SVA VLPs vaccine.
Subject(s)
Animals , Antibodies, Neutralizing , Escherichia coli/genetics , Genomics , Guinea Pigs , Picornaviridae/geneticsABSTRACT
BACKGROUND: Placenta and fetal membrane play an important role In maternal-fetal homeostasis. However, the molecular and cellular mechanisms underlying water transfer across placenta and amniotic membrane remain unknown. It is hypothesized that maternal-fetal fluid exchanges via aquaporin (AQP) water channels in the placenta and fetal membrane.OBJECTIVE: To investigate AQP8 protein expression in normal human placenta and fetal membrane.DESIGN, TIME AND SETTING: A control observation was performed at the Central Laboratory of Guangzhou Medical College from July to December 2005.MATERIALS: Human placenta and fetal membrane tissues from 5 elective cesarean section deliveries of normal term pregnancies (range 37-42 weeks) were studied. Maternal age averaged (27?) years old. Experimental protocol was approved by the Hospital's Ethics Committee.METHODS: Thirty minutes after delivery, fetal membrane and placenta were dissected and washed with sterile physiological saline. Some were frozen at -80?, and the remaining tissues were fixed for 24-48 hours with 10% neutral formalin and paraffin embedded for immunohistochemical staining.MAIN OUTCOME MEASURES: AQP8 expression and distribution in human placenta and fetal membrane were detected by the reverse transcription-polymerase chain reaction (RT-PCR), immunohistochemistry, and Western blotting analysis.RESULTS: RT-PCR results showed that AQP8 mRNA was expressed in both placenta and fetal membrane tissues. Western blotting analysis also yielded positive results in placenta and fetal membrane with a specific band site at approximately 45 000.Immunohistochemistry results revealed that AQP8 protein was expressed in placental syncytiotrophoblasts, amniotic epithelial cells, and chorion cytotrophoblasts.CONCLUSION: At protein level, AQP8 is expressed in placental syncytiotrophoblasts, amniotic epithelial cells, and chorion cytotrophoblasts.
ABSTRACT
Objective To determine the expression of aquaporin-1,3,8,9 mRNA (AQP-1,3,8, 9) in amniotic membranes in pregnant women with polyhydramnios. Methods Amniotic membranes were collected from women who presented with either polyhydramnios (n= 5)or normal amniotic fluid volume (control, n= 5) underwent elective cesarean sections at term. The AQP-1,3,8,9 mRNA expression were detected by reverse transcriptase-polymerase chain reaction (RT-PCR). Results The expression of AQP-9 mRNA on fetal membranes were significantly higher in polyhydramnios groups (1. 1403±0. 0831) than that of control (0. 5903±0. 1909) (P = 0. 002), although the expression of AQP-1, 3 and 8 showed no significant difference between the two groups (P= 0. 972, 0. 242,0. 608, respectively). Conclusions AQP-9 may play an important role in maintaining the volume and the balance of different components of amniotic fluid in polyhydramnios cases.