ABSTRACT
Background: Spermatogenesis is a tightly regulated developmental process of male germ cells. The stages in spermatogenesis are mitosis, meiosis and spermiogenesis. One of the genes playing a role in meiosis is Cell Division Cycle 25A [CDC25A]. De-creased expression of CDC25A is associated with failure of spermatogenesis and sperm retrieval. Infertility examination for azoospermia has been limited on histolog-ical examination. Hence, molecular research to find marker genes for infertility will improve the examination of testis biopsies
Methods: This research is a cross sectional study of 50 testicular biopsies with John-sen scoring categories from scoring 2 to 8. Analysis of mRNA expression used qPCR and protein expression using immunohistochemistry. Statistical analysis with Spear-man correlation was considered significant at p<0.05
Results: The result showed that transcript level and protein expression of CDC25A decreased in score 5 of Johnsen scoring categories. Moderate Spearman rho correla-tion [r=0.546] between mRNA relative expression and protein expression of CDC25A was significant at p<0.01
Conclusion: Decreased expression of CDC25A is associated with meiotic arrest as the etiology of spermatogenic failure in many azoospermic men
ABSTRACT
Background: Recently, Phosphatidylcholine [PC] has been used as an off-label treatment for lipolysis injection, which is associated with inflammatory reaction due to sodium deoxycholate, an emulsifier, so that inflammation as side effect occurs in those patients. Liposome formulation from soybean lipid was thought to be a better and safer alternative. This study aimed to analyze the mechanism of Liposomal Soybean Phosphatidylcholine [LSPC] extract from Indonesian soybeans [containing 26% PC] to induce Adipose-derived Stem Cells [ASCs] death in vitro
Methods: Liposomes were prepared using thin film hydration method followed by a stepwise extrusion process to produce a small amount of 41.0-71.3 nm. Liposomal soybean phosphatidylcholine extract [LSPCE], liposomal purified PC [LPCC], and solution of PC+SD were used for comparison. Annexin V-fluorescein Isothiocyanate/ Propidium Iodide [FITC/PI] double staining by flow cytometry and also measurement of caspase-3 activity using ELISA were used to quantify the rate of apoptosis. ASCs viability was measured using MTT assay after induction with liposomes. Morphological changes were shown using a phase-contrast, inverted microscope and Transmission- Electron Microscope [TEM]
Results: The flow cytometry results showed that cells treated with both LSPCE and LPCC showed increase in early apoptosis beginning at 6 hr after incubation, which was confirmed by caspase 3 measurement. MTT assay showed that both LSPCE and LPCC could decrease viability of cells. Cells treated with LSPCE and LPCC showed some rounded cells, which was an early sign of cell death. Cells treated with SD showed extensive membrane damage with necrosis features using TEM
Conclusion: The results above demonstrated that LSPCE induced apoptosis of ASCs