The immunoregulatory property of mesenchymal stem cells in Crocin treatment by expression modulation of microRNA-155, microRNA-21, microRNA-23b, microRNA-126a, and their target inflammatory genes.

Mesenchymal stem cells (MSCs) have high priority in clinical applications for treatment of immune disorders because of their immunomodulatory function. A lot of researches have currently been undertaken to enhance the stemness capacities of the cells and pick an excellent type of MSCs for clinical approaches. This study aims to assess the immunomodulatory related MicroRNAs (miRNAs) expression as well as their target genes in both adipose derived stem cells (Ad-SCs) and dental pulp derived stem cell (DP-SCs) in the presence or lack of Crocin (saffron plant's bioactive compound). For this purpose, first MSCs were extracted from adipose and dental pulp tissues, and then their mesenchymal nature was confirmed using flow cytometry and differentiation tests. Following the cell treatment with an optimal-non-toxic dose of Crocin (Obtained by MTT test), the expression of 4 selected immunomodulatory-related micro-RNAs (Mir-126, -21, -23, and-155) and their target genes (PI3K/ Akt 1 and 2/ NFKB and RELA) were assessed by RT-PCR. Our findings revealed that miRNA-23 and miRNA-126 were up-regulated in both types of cells treated with Crocin, while in the other side, miRNA-21 and miRNA-155 were down-regulated in DP-SCs and were up-regulated in Ad-SCs under treatment. Moreover, the real-time PCR results indicated that Crocin could significantly down regulate the expression of PI3K/ Akt1/ Akt2/ NFKB/ RELA genes in DP-SCs and PI3K/Akt2 genes in Ad-SCs and up regulate the expression of Akt1/ NFKB/ RELA genes in recent cells. Based on the analysis of the obtained data, the immunoregulatory effects of Crocin were higher in DP-SCs than in Ad-SCs. In conclusion, Crocin could control essential signaling pathways related to the inflammation by regulating the expression of related- miRNAs genes that play a key function in the immune regulation pathways in MSCs. Our findings can give an understanding of the mechanisms by which Crocin enhances the immunomodulatory feature of MSCs. According to the research findings, DP-SCs are probably a better immunomodulator in Crocin treatment than Ad-SCs and it may be helpful for MSCs selection in clinical applications for modulation or treatment of autoimmune disorders.

Cefazolin-Loaded Double-Shelled Hollow Mesoporous Silica Nanoparticles/Polycaprolactone Nanofiber Composites: A Delivery Vehicle for Regenerative Purposes.

Purpose: As important challenges in burn injuries, infections often lead to delayed and incomplete healing. Wound infections with antimicrobial-resistant bacteria are other challenges in the management of wounds. Hence, it can be critical to synthesize scaffolds that are highly potential for loading and delivering antibiotics over long periods. Methods: Double-shelled hollow mesoporous silica nanoparticles (DSH-MSNs) were synthesized and loaded with cefazolin. Cefazolin-loaded DSH-MSNs (Cef*DSH-MSNs) were incorporated into polycaprolactone (PCL) to prepare a nanofiber-mediated drug release system. Their biological properties were assessed through antibacterial activity, cell viability, and qRT-PCR. The morphology and physicochemical properties of the nanoparticles and nanofibers were also characterized. Results: The double-shelled hollow structure of DSH-MSNs demonstrated a high loading capacity of cefazolin (51%). According to in vitro findings, the Cef*DSH-MSNs embedded in polycaprolactone nanofibers (Cef*DSH-MSNs/PCL) provided a slow release for cefazolin. The release of cefazolin from Cef*DSH-MSNs/PCL nanofibers inhibited the growth of Staphylococcus aureus. The high viability rate of human adipose-derived stem cells (hADSCs) in contact with PCL and DSH-MSNs/PCL was indicative of the biocompatibility of nanofibers. Moreover, gene expression results confirmed changes in keratinocyte-related differentiation genes in hADSCs cultured on the DSH-MSNs/PCL nanofibers with the up-regulation of involucrin. Conclusion: The high drug-loading capacity of DSH-MSNs presents these nanoparticles as suitable vehicles for drug delivery. In addition, the use of Cef*DSH-MSNs/PCL can be an effective strategy for regenerative purposes.

Thymoquinone enhanced the antitumor activity of cisplatin in human bladder cancer 5637 cells in vitro.

PURPOSE: Cisplatin-based chemotherapy is a primary alternative for treating bladder cancer. But drug resistance and various side effects are the main unsightliness challenges. In search of a novel chemotherapeutic approach, this study was conducted to investigate whether thymoquinone (TQ) chemosensitize 5637 bladder cancer cells to cisplatin (CDDP). METHODS: The IC50 for each drug was first determined. The cells were then pre-exposed to 40 µM of TQ for 24 h before being treated with 6 µM of cisplatin. The viability and the sub-G1 population of the 5673 cells were respectively evaluated by alamar blue assay and propidium iodide staining. RT-qPCR was also applied to analyze the expression profile of the apoptosis-related genes (Bax, Bcl-2, p53). RESULTS: The viability of the cells treated with the combination of TQ and CDDP was significantly decreased compared to CDDP- or TQ-treated cells. TQ at the concentration of 40 µM increased the cytotoxicity of 6 µM CDDP by 35.5%. Moreover, flow cytometry analysis indicated that TQ pre-treatment of the cells resulted in a 55.5% increase in the population of 5637 cells in the sub-G1 phase compared to cells treated with CDDP alone. The results from RT-qPCR exhibited that the exposure of the cells to both TQ and CDDP significantly elevated Bax/Bcl-2 ratio by down-regulating Bcl-2 expression. CONCLUSION: TQ significantly increased the cytotoxicity of CDDP in 5637 cells and induced apoptosis by down-regulation of the Bcl-2. Therefore, TQ and CDDP might be an effective therapeutic combination for TCC bladder cancer treatment.

Identification of Potential Prognostic Markers and Key Therapeutic Targets in Hepatocellular Carcinoma Using Weighted Gene Co-Expression Network Analysis: A Systems Biology Approach.

Background: As the most prevalent form of liver cancer, hepatocellular carcinoma (HCC) ranks the fifth highest cause of cancer-related death worldwide. Despite recent advancements in diagnostic and therapeutic techniques, the prognosis for HCC is still unknown. Objectives: This study aimed to identify potential genes contributing to HCC pathogenicity. Materials and Methods: To this end, we examined the GSE39791 microarray dataset, which included 72 HCC samples and 72 normal samples. An investigation of co-expression networks using WGCNA found a highly conserved blue module with 665 genes that were strongly linked to HCC. Results: APOF, NAT2, LCAT, TTC36, IGFALS, ASPDH, and VIPR1 were the blue module's top 7 hub genes. According to the results of hub gene enrichment, the most related issues in the biological process and KEGG were peroxisome organization and metabolic pathways, respectively. In addition, using the drug-target network, we discovered 19 FDA-approved medication candidates for different reasons that might potentially be employed to treat HCC patients through the modulation of 3 hub genes of the co-expression network (LCAT, NAT2, and VIPR1). Our findings also demonstrated that the 3 scientifically validated miRNAs regulated the co-expression network by the VIPR1 hub gene. Conclusion: We found co-expressed gene modules and hub genes linked with HCC advancement, offering insights into the mechanisms underlying HCC progression as well as some potential HCC treatments.

Anti-Inflammatory Activity of Ferula assafoetida Oleo-Gum-Resin (Asafoetida) against TNF-α-Stimulated Human Umbilical Vein Endothelial Cells (HUVECs).

Inflammation is the body's biological reaction to endogenous and exogenous stimuli. Recent studies have demonstrated several anti-inflammatory properties of Ferula species. In this paper, we decided to study the anti-inflammatory effect of ethanolic extract of Ferula assafoetida oleo-gum-resin (asafoetida) against TNF-α-stimulated human umbilical vein endothelial cells (HUVECs). HUVECs were cultured in a flat-bottom plate and then treated with ethanolic extract of asafoetida (EEA, 0-500 µg/ml) and TNF-α (0-100 ng/ml) for 24 h. We used the MTT test to assess cell survival. In addition, the LC-MS analysis was performed to determine the active substances. HUVECs were pretreated with EEA and then induced by TNF-α. Intracellular reactive oxygen species (ROS) and adhesion of peripheral blood mononuclear cells (PBMCs) to HUVECs were evaluated with DCFH-DA and CFSE fluorescent probes, respectively. Gene expression of intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), and E-selectin and surface expression of ICAM-1 protein were measured using real-time PCR and flow cytometry methods, respectively. While TNF-α significantly increased intracellular ROS formation and PBMC adhesion to TNF-α-induced HUVECs, the pretreatment of HUVECs with EEA (125 and 250 µg/ml) significantly reduced the parameters. In addition, EEA pretreatment decreased TNF-α-induced mRNA expression of VCAM-1 and surface protein expression of ICAM-1 in the target cells. Taken together, the results indicated that EEA prevented ROS generation, triggered by TNF-α, and inhibited the expression of VCAM-1 and ICAM-1, leading to reduced PBMC adhesion. These findings suggest that EEA can probably have anti-inflammatory properties.

Bentonite Nanoparticles and Honey Co-Administration Effects on Skin Wound Healing: Experimental Study in the BALB/c MICE.

In recent years, nanotechnology and the subsequent production of nanoparticles have developed excellent methods for medical applications, including wound healing. One of these nanoparticles is bentonite nanoparticles (BNPs) which show high ability in tissue engineering. But our knowledge of its effectiveness in wound healing is based on little data. Therefore, the main purpose of this study was to evaluate the wound healing ability of BNPs and in the next step the suitability of honey as a solvent for these nanoparticles. Methods: In this experimental study, an excisional wound injury model was developed in adult male BALB/c mice (n = 60) by creative two equal-sized wounds (5 mm) on either side of their back midline. The animals were allocated into five groups (n = 12 each) as untreated control (U), honey (H), polyethylene glycol (P), and (BNPs) dissolved in honey or polyethylene glycol (H + BNPs, P + BNPs). Animals have received their relative topical treatments twice per day for 14 consecutive days. Tissue sampling was carried out on days 4, 7, 10, and 14. The tissue sections were stained with hematoxylin and eosin and Trichrome-Masson staining methods. The histomorphological parameters including inflammatory cells infiltration, fibroblasts, re-epithelialization, granulation formation, and collagenases were evaluated in all tissue sections. Data were analyzed by SPSS 16 software. Comparison between the groups was performed by one-way analysis of variance following Tukey's post-hoc test. Compared to the control group, BNPs showed significant wound healing activities with lower inflammatory cells infiltration, higher fibroblastosis and new epithelium thickness, and greater granulation area and collagen fibers density in the ulcer bed. In addition, honey as a solvent synergistically increased the wound healing activity of the studied nanoparticle. These results for the first time are clearly showing that BNPs have a promising wound healing activity, especially when applied with honey concurrently.

miR-27 and miR-124 target AR coregulators in prostate cancer: Bioinformatics and in vitro analysis.

The inadequate efficacy of the current treatments for metastatic prostate cancer has directed efforts to the discovery of novel therapies. MicroRNAs (miRNAs) have been considered potential therapeutic agents due to their ability to control gene expression and cellular pathways. The accurate identification of genes and pathways which are targeted by a miRNA is the first step in the therapeutic use of these molecules. In this regard, there are multiple experimental and computational methods to predict and confirm the miRNA-mRNA relationships. The targeting the androgen receptor (AR) indirectly as the most important mediator of prostate cancer has been posited to both control the disease and prevent resistance to treatment. This study aimed to identify miRNAs targeting AR coregulators. For this purpose, we examined target genes by combining miRNA-mRNA computational and experimental data from various databases. miR-27a-3p and miR-124 displayed the highest scores and were selected as miRNAs with the potential to target candidate genes. Next, three cell lines of prostate cancer including PC3, LNCAP, and DU145 were transfected with plasmids which were expressed these selected miRNAs. Then, the gene expression and cell cycle analysis were performed. A decrease was observed in cell viability in all three cell lines than the cells transfected with backbone plasmid. Furthermore, the findings indicated that miR-27a-3p and miR-124 led to a significant decrease in the expression of all genes that were studied in PC3 cell line. In addition, miR-124 caused significant the cellular arrest in the G0/G1 stage, while for miR-27a-3p, this arrest occurred was in the G2/M stage. Our results indicated that the function of a unique miRNA could be different in different cell lines with particular cancer phenotype based on the cell line stage. These findings offer the possibility of employing the miR-124 and miR-27a-3p as therapeutic agents for prostate cancer treatment.

Wound Healing Effects of Chitosan Nanosheets/Honey Compounds in Male BALB/c Mice.

Background and aim: Up to now, proper wound care management has remained as an important clinical challenge. Chitosan nanosheets (CNSs) showed a great potential in tissue engineering, but our knowledge about their wound healing effectiveness is based on very limited data. Thus, the aim of this research was to evaluate the wound healing potential of CNSs and honey as a vehicle for these nanoparticles. Methods: The skin excisional wound injury model was made in adult male BALB/c mice (n = 60) by creating two identical sized wounds (5mm) on either side of their dorsal midline. The animals were divided into five groups (n = 12 each) as untreated control, honey, polyethylene glycol, and CNSs dissolved either in honey or polyethylene glycol. Animals were received their relative topical treatments twice per day for 14 consecutive days. Tissue sampling was carried out on days 4, 7, 10, and 14 post wounding. The histological parameters including inflammatory cells infiltration, fibroblast proliferation, re-epithelialization, granulation formation, and collagen formation were evaluated in all studied time points. Results: Compared to the control group, CNSs showed significant wound healing activities with lower inflammatory cells infiltration, higher fibroblastosis and new epithelium thickness, and greater granulation area and collagen fibers density in the ulcer bed. In addition, honey synergistically increased the wound healing activity of the studied nanoparticles. Conclusion: These results showed that CNSs have promising wound healing activity specially when dissolved with honey concurrently.

Fabrication and characterization of jujube extract-loaded electrospun polyvinyl alcohol nanofiber for strawberry preservation.

Recently, using of natural ingredients gains much attention in the field of food science and active packaging. In this study, first, jujube extract was investigated for its antimicrobial and antioxidant properties, and then, the effect of electrospun PVA/JE (jujube extract loaded into Poly vinyl alcohol) nanofiber as active packaging was evaluated to increase the shelf-life of strawberry. PVA/ZE nanofiber film was prepared using electrospinning method, and their morphology was confirmed by scanning electron microscopy (SEM). Fruit preservation abilities of the nanofiber film were tested on strawberries. The strawberries were then kept at 4℃ for 15 days and characterized in terms of their properties (weight loss, TSS, firmness, and sensory analysis). Results indicated that flavonoid content of jujube extract ranged from 4.80 ± 0.01 to 13.54 ± 0.08 mg CEQ/100 g, and the DPPH free radical-scavenging activity was from 210 ± 2.66 to 1498 ± 2.65 (GAE/g DW). The jujube extract also presented potent antibacterial activity against the investigated bacteria and fungi. The scanning electron microscopy (SEM) images of nanofibers had a linear morphology and bead-free structure; however, PVA/JE (jujube extract encapsulated into PVA nanofiber) had strip and flat organization. Strawberries in control group showed signs of decay and a decrease in visual appearance on the 6th. However, fruits in PVA/JE group had acceptable overall appearance for marketing, as no obvious sign of decay was observed on 12th day of storage. Active packaging containing herbal extracts and essential oils preserves the organoleptic and physicochemical properties of the fruits.

Recombinant Production of Bovine Enteropeptidase Light Chain in SHuffle® T7 Express and Optimization of Induction Parameters.

Enteropeptidase is a duodenum serine protease that triggers the activation of pancreatic enzymes by remarkably specific cleavages after lysine residues of peptidyl substrate (Asp)4-Lys. This high specific cleavage makes the enzyme a widely used biotechnological tool in laboratory researches and industrial scale. Previous studies both in small and large scales were showed low expression and miss-folding of the expressed protein. In this study, the DNA sequence encoding the light chain (catalytic subunit) of bovine enteropeptidase (EPL) was subcloned into plasmid pET-32b, downstream to the DNA encoding the fusion partner thioredoxin immediately after the EPL cleavage site. SHuffle® T7 Express was selected as an expression host due to the ability to promote proper folding and correction of the mis-oxidized bonds. Expression and purification of protein was performed, and the result of biological activity confirmed that the active EPL was obtained. Optimization of protein expression conditions was accomplished by response surface methodology for significant factors including induction temperature, duration of induction, inducer concentration and OD600 of induction. The best conditions were achieved in 1.05 mM IPTG at OD600 of 0.6 for seven h incubation at 26.5 °C, and a high level of protein expression was obtained in the optimized condition.

PD-L1 overexpression conveys tolerance of mesenchymal stem cell-derived cardiomyocyte-like cells in an allogeneic mouse model.

Although the autologously transplanted cells are immunologically durable, allogeneic cell transplantation is inevitable in a series of cases. Mesenchymal stem cells (MSCs) are one of the suitable candidates for cardiac tissue regeneration that have been shown to acquire immunogenicity concurrent with cardiomyogenic differentiation. The present study aimed to exploit PD-L1, as a key immunomodulatory checkpoint ligand to protect the MSCs-derived cardiomyocyte-like cells (CLCs) against the detrimental alloimmunity. Mouse bone marrow-derived MSCs were stably transduced to overexpress PD-L1. MSCs were in vitro differentiated into CLCs and the expressions of immunologic molecules were compared between MSCs and CLCs. The in vitro and in vivo allogeneic immune responses were also examined. The differentiated CLCs had higher expressions of MHC-I and CD80. Upon in vitro coculture with allogeneic splenocytes, CLCs caused more CD4+ and CD8+ T cell activation, lymphocyte proliferation, and interferon-γ (IFN-γ) release in comparison to MSCs. PD-L1 overexpression on CLCs decreased the activation of CD8+ T cells, proliferation of lymphocytes, and release of IFN-γ. The PD-L1-overexpressing CLCs elicited lower in vivo CD4+ and CD8+ T cell activation and reduced the anti-donor antibody response accompanied by increased durability and reduced T cell infiltration. The present study verified the potential of PD-L1 overexpression as a preparative strategy for the protection of allogeneic MSCs-derived CLCs against the detrimental alloreaction.

Effect of Pomegranate Extract in Mesenchymal Stem Cells by Modulation of microRNA-155, microRNA-21, microRNA-23b, microRNA-126a, and PI3K\AKT1\NF-[Formula: see text]B Expression.

Today, mesenchymal stem cells (MSCs) are candidates for various autoimmune disease treatments due to immunomodulatory activity in these cells. Much research has recently been done to improve the immunomodulatory activity of MSCs. Genetic variation is one of these methods. microRNAs (miRNAs) are small noncoding RNAs that control most of the cell's biological activities. Recent studies have shown that miRNAs play a significant role in the regulation of MSC immunomodulatory activity. Pomegranate is a fruit that has antioxidant, anti-inflammatory, and anticancer properties and has been used for many years for therapeutic purposes. The objective of this research is to evaluate the immunoregulatory-related miRNAs level of adipose-derived MSCs (Ad-MSCs) obtained from adipose tissue in the presence or lack of pomegranate (Punica granatum) extract (PGE). Our results showed that miRNA-23 and miRNA-126 were upregulated by PGE treatment in MSCs, and in contrast, miRNA-21 and miRNA-155 were downregulated by PGE treatment in MSCs. In addition this research shows that PGE can downregulate the expression of PI3K\AKT1\NF-[Formula: see text]B in Ad-MSCs. Our bioinformatics data have shown that the target of these four miRNAs and the signaling pathways, in which these targets are involved, can play an important role in regulating the immunomodulation function of stem cells. In conclusion, PGE can inhibit the expression of PI3K\AKT1\NF-[Formula: see text]B genes involved in inflammatory pathways via miRNA-23 and miRNA-126 overexpression or miRNA-21 and miRNA-155 downregulation that plays a role in the pathways of immune modulation in Ad-MSCs. These results may provide insight into the mechanism underlying the regulation of the immunomodulatory activity of Ad-MSCs by PGE.

Characterization and Release Behavior of a Thiosemicarbazone from Electrospun Polyvinyl Alcohol Core-Shell Nanofibers.

Mats of polyvinyl alcohol (PVA) core-shell nanofibers were produced using coaxial electrospinning in the presence of a thiosemicarbazone (TSC) N4-(S)-1-phenylethyl)-2-(pyridin-2-yl-ethylidene)hydrazine-1-carbothioamide (HapyTSCmB). Monolithic fibers with 0% or 5% TSC and core-shell fibers with 10% TSC in the spinning solution were studied to compare stability and release rates. SEM showed the formation of uniform, bead-free, cylindrical, and smooth fibers. NMR spectroscopy and thermal analysis (TG/DTA) gave proof for the chemical integrity of the TSC in the fiber mats after the electrospinning process. Attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy showed no TSC on the surface of the PVA/TSC-PVA fibers confirming the core-shell character. The TSC release profiles of the fibers as studied using UV-vis absorption spectroscopy showed a slower release from the PVA/TSC-PVA core-shell structure compared with the monolithic PVA/TSC fibers as well as lower cumulative release percentage (17%). Out of several release models, the Korsmeyer-Peppas model gave the best fit to the experimental data. The main release phase can be described with a Fick-type diffusion mechanism. Antibacterial properties were tested against the Gram-positive Staphylococcus aureus bacterium and gave a minimal inhibitory concentration of 12.5 µg/mL. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazoliumbromide (MTT)-based cytotoxicity experiments showed that the cell viability of fibroblast at different contents of TSC was slightly decreased from 1.5% up to 3.5% when compared to control cells.

Jujube and green tea extracts protect human fibroblast cells against UVB-mediated photo damage and MMP-2 and MMP-9 production.

OBJECTIVE: Oxidative stress and ultraviolet B (UVB) irradiation are known as principal inducers of DNA damage and modulators of gene expression in aging process and skin photoaging, which are associated with upregulation of matrix metalloproteinases (MMPs). Because of the antioxidant capacity of jujube and green tea, we decided to determine their protective effects of human fibroblast cells against UVB-induced photo-damage and reduction of MMP-2 and MMP-9 expression. MATERIALS AND METHODS: We exposed human fibroblast cells to different doses of UVB (0-20 mJ/cm2) with or without different concentrations of jujube and green tea extracts. Cell viability was assessed using MTT assay. Total antioxidant capacity and free radical scavenging activity of cell supernatant were assessed using FRAP and DPPH methods, respectively. The concentrations of MMP-2 and MMP-9 in the samples were determined by ELISA kits. RESULTS: Fibroblast cells viability, 24 hr after UVB irradiation, reduced about 70% compared to the controls. Pre-treatment of the cells with jujube extract (8 mg/ml) increased the cell viability by almost 85% while green tea (0.5 mg/ml) protected the irradiated cells by 71%. Also, MMP-2 and MMP-9 content decreased in a concentration-dependent manner in the cells pre-treated with jujube and green tea extracts. CONCLUSION: These data suggest that jujube and green tea could be useful to attenuate solar UVB light-induced oxidative stress and skin photoaging and can be suggested as a potential candidate for the development of new anti-UVB medicines and cosmetic products.

Immunoregulatory properties of mesenchymal stem cells: Micro-RNAs.

Mesenchymal stem cells (MSCs) are multipotent cells that are excellent candidates for different cellular therapies due to their physiological properties such as immunoregulatory function. whetheare currently utilized for regenerative medication and treatment of a number of inflammatory illnesses given their ability to considerably impact tissue microenvironments via extracellular vesicles or toll-like receptor pathway modulation. MicroRNAs (miRNAs) are small noncoding RNAs that target the messenger RNA and play a critical role in different biological procedures, such as the development and reaction of the immune system. Moreover, miRNAs have recently been revealed to have serious functions in MSCs to regulate immunomodulatory properties. In this review, we study how the miRNAs pathway can modulate the immunoregulatory activity of MSCs by counting their interactions with immune cells and also discuss the possibility of using miRNA-based implications for MSC-based therapies.

Cytoprotective effects of the aqueous extract of the Ziziphus jujuba fruit on TBHP-induced damage on human fibroblast cells.

Background Oxidative stress plays a major role in the development of various human diseases. However, many antioxidant compounds can neutralize the excess of free radicals, protect the cells against their toxic effects and help prevent or treat a disease. This study investigated the cytoprotective effects of the aqueous extract of the Ziziphus jujuba fruit on the tert-butyl hydroperoxide (TBHP)-induced damage on human fibroblast cells. Methods Human fibroblast cells were pretreated with different concentrations (1, 2, 4, 8 mg/mL) of Z. jujuba for 24 h and exposed to 75 µM TBHP for another 24 h. Cell viability was determined by the MTT assay. The antioxidant activity was determined using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) methods, and the intracellular antioxidant activity was evaluated with the Cellular antioxidant activity assay. Results Our data showed that treatment with TBHP reduced cell viability of human fibroblast cells, while pretreatment with Z. jujuba increased cell viability in a dose-dependent manner. This indicated the cytoprotective effects of Z. jujuba. Pretreatment with Z. jujuba increased the antioxidant capacity and scavenged the TBHP-produced peroxyl radicals in the human fibroblast cell medium. Moreover, Z. jujuba increased the intracellular antioxidant activity of human fibroblast cells. Conclusions These results demonstrated that the aqueous extract of the Z. jujuba fruit can prevent TBHP-induced cellular toxicity by enhancing the antioxidant activity in cells and their medium. So, Z. jujuba has a therapeutic potential to attenuate oxidative stress-induced diseases.

Prevention of kidney cell damage in hyperglycaemia condition by adiponectin.

Adiponectin (APN) is an adipocytokine, secreted from adipose tissue and has anti-inflammatory, anti-ageing, and antidiabetic properties. Hyperglycaemia can damage the renal cells, and mammalian target of rapamycin (mTOR), along with Sirtuin 1 (SIRT1), have an important role in kidney cell response to hyperglycaemia. Therefore, understanding the relationship between adiponectin, mTOR, and SIRT1 proteins is beneficial for deciphering the mechanism of adiponectin function. In this study, Human Embryonic Kidney-293 (HEK-293) cells were cultured under normal and high-glucose condition, with and without APN (1, 10, and 100 ng/mL) for 48 hours. mTOR protein expression was evaluated by western blot analysis, and SIRT1 protein was assessed using ELISA method. To evaluate hyperglycaemia-mediated cytotoxicity, cell viability was determined using MTT assay. Data showed that APN in high dose (100 ng/mL) significantly reduced the expression of mTOR and p-mTOR, increased SIRT1 protein, and also improved cell viability compared with the control high glucose (p ≤ 0.05). According to this results, APN can be useful in preventing renal cell damage, by affecting on the expression of mTOR and SIRT1 proteins, as well as increasing the survival of kidney cells in hyperglycaemia conditions. SIGNIFICANCE OF THE STUDY: Adiponectin triggered mTOR/p-mTOR/SIRT1 pathway and decreased cell death in human kidney cells. Our findings provide preliminary experimental data that support further studies on the potential therapeutic role of adiponectin in diabetes and diabetic-induced metabolic complications.

Adverse effects of BCG vaccine 1173 P2 in Iran: A meta-analysis.

Although in the last two decades the World Health Organization (WHO) has introduced tuberculosis as "a threat to global", the vaccination with the Mycobacterium bovis Bacillus Calmette-Guerin (BCG) is the only way for the prevention of this fatal infectious disease. Despite of the efficacy of BCG vaccine especially against infants' meningitis, it has still some limitations due to a variety of adverse effects. Many studies have evaluated the side effects of different strains of BCG vaccines in different countries. In Iran, some studies have been done so far to evaluate the adverse effects of 1173 P2 strain which is used for BCG vaccination. Each of these studies have used different standardization and sampling methods. This review will survey all studies that have been published about adverse effects of 1173 P2 strain of BCG vaccine in Iran using data mining methods.

Antiproliferative and Proapoptotic Effects of Crocin Combined with Hyperthermia on Human Breast Cancer Cells.

We investigated the suppressive effects of crocin alone and in combination with hyperthermia (HT) on proliferation of breast cancer cells. Cell viability, colony formation ability, and apoptosis were assessed by 3-(4,5-dimetylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide (MTT), soft agar, Hoechst 33258 staining, and percentage of lactate dehydrogenase (LDH) release methods, respectively. The mRNA levels Hsp27, Hsp70, Hsp90, Bax, and Bcl-2 were measured by quantitative real-time polymerase chain reaction (qRT-PCR). Hsp70 and Hsp90 proteins were determined using enzyme-linked immunosorbent assay (ELISA) technique. Crocin in combination with HT significantly inhibited the proliferation of cancer cells in a dose- and time-dependent manner. There was a degree of synergism in the combined treatment. However, crocin did not show the high cytotoxic effect on normal cells. This treatment decreased colony formation of cancer cells up to 94%. Changed nuclear morphology and increased LDH indicated that crocin combined with HT has a more apoptotic effect than crocin alone. Furthermore, in treated cells Bax/Bcl-2 ratio markedly increased, whereas expression of heat-induced genes decreased. Also, the Hsp70 and Hsp90 proteins decreased in the treated cells. Our study indicated that combination of crocin and HT has strong antiproliferative and apoptotic activities against breast cancer cells. Hence, it is suggested that more studies are warranted to apply crocin as a possible, safe, and promising anticancer agent in cancer.

Regulation of connexin 43 and microRNA expression via ß2-adrenoceptor signaling in 1321N1 astrocytoma cells.

Connexin 43 (Cx43) is the main gap junction protein in astrocytes and exerts the same effects on growth inhibition in astrocytoma and glioma as microRNA-146a (miR-146a) in glioma. ß2-adrenergic receptor (AR) signaling modulates Cx43 expression in myocytes via components downstream of protein kinase A (PKA) and exchange protein directly activated by cAMP (Epac). However, it remains to be elucidated how expression of Cx43 is modulated in astrocytes. In the present study, 1321N1 astrocytoma cells were treated with ß2-AR signaling agents in order to evaluate the expression of Cx43 and miRNAs. RNA and protein were extracted from the cells for use in reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The results revealed that clenbuterol increased miR-146a level and upregulated Cx43 expression via cAMP/PKA at the mRNA and protein level. Pre-inhibition of adenyl cyclase decreased expression of Cx43 and miR-146a. PKA activation and overexpression of miR-146a in A-1321N1 cells increased the expression of Cx43. ß2-AR stimulation and 6Bnz, a PKA activator, suppressed oncomiRs miR-155 and miR-27a, while 8-(4-chlorophenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate, an Epac activator, increased their levels. The current findings demonstrated that ß2-AR signaling has growth inhibitory effects via modulation of the cAMP/PKA pathway in A-1321N1 cells through increasing the expression level of Cx43 and miR-146a as well as decreasing miR-155 and miR-27a levels. Thus, stimulation of the ß2-AR and PKA signaling pathway may be a useful approach for astrocytoma therapy.