cellular uptake of antisense oligonucleotid of E6 mRNA into cervical cancer cells by DOPE-modified hydroxyapatite nanoparticles

Although several chemical and physical methods for gene delivery have been introduced, their cytotoxicity, non-specific immune responses and the lack of biodegradability remain the main issues. In this study, hydroxyapatite nanoparticles [NPs] and 1, 2-dioleoyl-sn-glycero-3-phosphoethanolamine [DOPE]-modified hydroxyapatite NPs was coated with antisense oligonucleotide of E6 mRNA, and their uptakes into the cervical cancer cell line were evaluated. Calcium nitrate and diammonium phosphate were used for the synthesis of the hydroxyapatite nanoparticle. Thus, they were coated with polyethylene glycol [PEG], DOPE and antisense oligonucleotide of E6 mRNA using a cross-linker. Then, hydroxyapatite NPs and DOPE-modified hydroxyapatite NPs were incubated 48 hours with cervical cancer cells and their uptakes were evaluated by fluorescent microscopy. The hydroxyapatite NPs had different shapes and some agglomeration with average size of 100 nm. The results showed DOPE-modified hydroxyapatite NPs had higher uptake than hydroxyapatite NPs [P<0.05]. Hydroxyapatite NPs conjugated with DOPE are a good choice for gene delivery and silencing of viral genes in cervical cancer cells, but their efficacy should be addressed more in future studies

Detection of chalmydia trachomatis infection in female partners of infertile couples

The prevalence of infertility is about 10-15% among the couple in reproductive age. Several factors can affect on fertility ability in men and women. Chlamydia is non-motile gram negative obligatory interacellular pathogenic organism. It can be caused infections in females as cervicitis, urethritis, endometritis, pelvic inflammatory disease also prostatitis and epidydidimitis in male as well. The aim of this survey is to mention the frequency of infection with Chlamydia infection in infertile female who were treated in Yazd research and Clinical Centre for Infertility. A questionnaire contains some demographic information and a clinical feature related to the infection was completed for each infertile women. Specimen of vaginal discharge was collected by well trained nurses using sterile cotton swap from 91 women. Elisa test was done on blood serum. DNA extraction for Chlamydia was carried out using low salt method and PCR was done using MOMP and plasmid primers. DNA sequencing was performed on two PCR products using Chromas LITE ver. 2.01 and analyzed by BLAST. Of 91 blood samples collected in this survey, none of those was positive by ELISA. Also there was not positive by PCR. Four PCR products showed a questionable band which were not in range of Chlamydia. The products were performed DNA sequencing and there were not related to any micro-organism. However, it is well known that Chlamydia has an infection role in infertility. Nevertheless, there was not evidence of this organism in these infertile patients. It is necessary to design such survey in large population of infertile patients especially on infertile women with tubal infertility and their husbands as well