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Chitosan (Cs) was extracted from shrimp shell and its derivative forms including N-alkyl (AlkCs) and nanoparticles (CsNPs) were prepared. First, the properties of nanoparticles were determined by dynamic light scattering (DLS) and the morphology of nanoparticles and N-alkylated by scanning electron microscopy (SEM). Then their antibacterial activity was evaluated by the test of minimum inhibitory (MIC) and lethal (MBC) concentration, diffusion on agar by disk, permeability of cell membrane by measurement of cytoplasmic beta-galactosidase release (ONPG). The type of apoptosis cell death was also examined by DAPI staining and changes in cell surface integrity by atomic force microscopy (AFM). The results showed that the nanoparticles are spherical with an average hydrodynamic diameter of 240 nm. N-alkyl had a rough surface structure compared to native chitosan. At the least of MIC (78 μg/ml) and MBC (100 μg/ml) points were observed for CsNPs (P < 0.05). Nanoparticles and N-alkyl of chitosan showed the highest diameter of growth inhibition zone at 1250 concentration compared to other disks (p <0.05). Outer membrane permeability of derivative forms of chitosan showed significant differences with native chitosan and cells of control. DAPI staining test showed higher cell death of chitosan-derived forms. DAPI staining test showed higher cell death of derivative of chitosan. The images obtained from AFM showed a change in the membrane integrity of the treated cells compared to spherical and clustered of control cells. Thus, the antibacterial properties of native chitosan improved by physical and chemical modification.
 

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SH-SY5Y is a neuroblastoma cell line which used as a cancer and neurodegenerative disorders model and its neuro-experimental studies. The different diseases cause by a defect in apoptosis pathway. Disruption of apoptotic proteins has an effect on the treatment process and response to drugs. In nerve cells, due to the high expression of apoptosis inhibitory proteins, the efficacy of drugs is low. Combination therapy is one of the developing treatment methods. The aim of this research is to evaluate the effectiveness of doxorubicin drug on apoptosis in SH-SY5Y cells under the conditions of high expression of caspase9. Caspase9 is a key enzyme in intrinsic apoptosis. First, cell viability was obtained through MTT assay under the different drug concentrations. Then, caspase9 gene was transfected in cells and affected by the concentration lower than IC50 of drug, and cell energy level and cell death were checked by different methods. ATP assay showed that the expression of caspase9 with drug lead to ATP decreases. Caspase3/7 activity indicated an increase in cell death by drug and caspase. Propidium staining to hoechst showed that the expression of caspase9 in combination with doxorubicin induce more death. To ensure the expression levels of protein that induces cell death, the amount of caspase3 protein was checked by western blotting, which showed a significant increase in combination of caspase9 and drug. Our findings showed that the induction of caspase9 expression intensifies the effect of drug and the combined treatment may be effective on the responsiveness of neuronal diseases.
 

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Objective: Breast cancer is the second leading cause of cancer death in women. Cisplatin is a traditional cancer drug commonly used in chemotherapy for killing cancer cells. Modulation at the mRNA levels of apoptotic related genes often correlate with the sensitivity of various types of cancer cells to chemotherapeutic agents. Nanoparticulate drug delivery systems are being developed to effectively deliver smaller doses of chemotherapeutic agents and control drug distribution in the body. In this study, we evaluate the expressions of BCL2 and BAX genes in T47D treated with cisplatin and cisplatin nanoparticles, which can result in a new approach to breast cancer therapy. Methods: In this study, we treated T47D cells with different concentrations of cisplatin and cisplatin nanoparticles at 48 h. The IC50 was determined. We extracted RNA by using RNX solution, after which cDNA was synthesized. The precise primers for the BCL2, BAX and TBP genes were designed by specific software. The quantity of BCL2 and BAX gene expression compared to TBP gene (reference gene) was analyzed using real-time PCR.  Results: BCL2 and BAX gene expression levels in T47D cells treated by cisplatin were 0.7 (BCL2) and 1.48 (BAX); in T47D cells treated with cisplatin-loaded nanoparticles, the gene expressions were 0.03 (BCL2) and 2.41 (BAX). Conclusion: In this study, the results have shown that cisplatin-loaded nanoparticles are effective anticancer agents. Cisplatin nanoparticles induce apoptosis in human breast cancer cell lines. We have shown that cisplatin nanoparticles strongly increased cytotoxicity in comparison to the free drug in the T47D cell line.

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Aims: The present study aimed to evaluate the developmental rate of ovarian follicles and the incidence of cell death in grafted immature mouse ovarian tissue encapsulated and non-capsulated in sodium alginate.
Materials and Methods: Female (NMRI) mice (n=50) were divided into 3 groups as follows: Group A; the right ovary was removed and encapsulated in sodium alginate then transplanted under kidney capsule, Group B; the right ovary was removed and without encapsulation transplanted under kidney capsule, in both transplanted groups the left ovary was intact. Group C; control group, both ovaries were intact. After transplantation, in the first and fourth estrous cycles at proestrus phase. The morphology of the grafted ovaries, and the percentage of normal follicles were evaluated using hematoxylin and eosin staining. The incidence of apoptosis cell death was evaluated by anti-BAX immunohistochemical staining.
Findings: At first and fourth estrous cycle, almost 99.5% of follicles had normal morphology and no significant difference was observed between the groups. The follicular development and growth rate in the two grafted groups, was significantly higher than the control group, moreover, these rates were higher in the capsulated group than non-capsulated once (p<0.05). In spite of the presence of some BAX positive cells in the preantral and antral follicles, there was no remarkable reaction for BAX antibody in the primordial and primary follicles in studied groups.
Conclusion: Despite the high developmental rat and premature ovarian reserve depletion in grafted groups that can affect the longevity of transplanted tissue, while sodium alginate has a positive effect on the follicular development in grafted tissue.