Differentiation of Wharton's Jelly-derived mesenchymal stem cells into insulin-producing beta cells with the enhanced functional level on electrospun PRP-PVP-PCL/PCL fiber scaffold.

Diabetes is a global problem that threatens human health. Cell therapy methods using stem cells, and tissue engineering of pancreatic islets as new therapeutic approaches have increased the chances of successful diabetes treatment. In this study, to differentiate Wharton's Jelly-derived mesenchymal stem cells (WJ-MSCs) into insulin-producing cells (IPCs) with improved maturity, and function, platelet-rich plasma (PRP)-Polyvinylpyrrolidone (PVP)-Polycaprolactone (PCL)/PCL scaffold was designed. The two-dimensional (2D) control group included cell culture without differentiation medium, and the experimental groups included 2D, and three-dimensional (3D) groups with pancreatic beta cell differentiation medium. WJ-MSCs-derived IPCs on PRP-PVP-PCL/PCL scaffold took round cluster morphology, the typical pancreatic islets morphology. Real-time PCR, immunocytochemistry, and flowcytometry data showed a significant increase in pancreatic marker genes in WJ-MSCs-derived IPCs on the PRP-PVP-PCL/PCL scaffold compared to the 2D-experimental group. Also, using the ELISA assay, a significant increase in the secretion of insulin, and C-peptide was measured in the WJ-MSCs-derived IPCs of the 3D-experimental group compared to the 2D experimental group, the highest amount of insulin (38 µlU/ml), and C-peptide (43 pmol/l) secretion was in the 3D experimental group, and in response to 25 mM glucose solution, which indicated a significant improvement in the functional level of the WJ-MSCs-derived IPCs in the 3D group. The results showed that the PRP-PVP-PCL/PCL scaffold can provide an appropriate microenvironment for the engineering of pancreatic islets, and the generation of IPCs.

Adipose-derived mesenchymal stem cells -conditioned medium effects on Glioma U87 cell line migration, apoptosis, and gene expression.

The conditioned medium of mesenchymal stem cells (MSCs) has controversial roles in cancer, either promoting or suppressing tumor growth. Our research on the results of adipose tissue-derived MSC (AD-MSC)-conditioned media on U87 glioma cells was motivated by the disputed role of mesenchymal stem cells (MSCs) in cancer, which may either promote or inhibit tumor growth. Using flow cytometry, AD-MSCs were identified, verified, and their conditioned media was used to treat U87 cells. Through RT-qPCR, scratch assay, and apoptosis analysis, we evaluated gene expression (SOX4, H19, and CCAT1), cell migration, and apoptosis in U87 cells.The conditioned media greatly increased the expression of SOX4 and H19, but not CCAT1. Although there were few differences in migration and apoptosis, both were slightly increased in the treated group.These outcomes have drawn attention to the complexity of the interactions between MSCs and glioma cells. This complexity requires further research to identify the specific mechanisms governing MSC-mediated impacts on the development of glioblastoma multiforme (GBM).

Effect of incorporating aluminum oxide nanoparticles on thermal conduction and flexural strength of acrylic resins.

Background: The mechanical and thermal properties of polymethyl methacrylate, as the most commonly used material for the fabrication of dental prostheses, should be improved due to its structural weaknesses. The present study aimed to compare the flexural strength and thermal conduction of two heat-cured and self-cured acrylic resins reinforced with aluminum oxide nanoparticles. Materials and Methods: In this in vitro study, a total of 114 samples consisting of heat- and self-cured three subgroups (1% and 3% Al2O3 and the control) with 66 samples for the thermal conduction (n = 11) and 48 samples for the flexural strength (n = 8) tests were prepared. Flexural strength was assessed with a three-point bending test using a universal testing machine. One-way ANOVA was applied for data analysis, followed by post hoc Tukey paired group comparison tests (P < 0.05). Results: An increase in the aluminum oxide nanoparticle percentage in acrylic resins increased the thermal conduction in heat-cured acrylic resin from 2.142 ± 0.0298 to 2.487 ± 0.0359 m (2)/sec and in self-cured acrylic resin from 2.0150 ± 0.02646 to 2.1475 ± 0.04031 m (2)/sec and decreased the flexural strength in heat-cured acrylic resin from 60.521 ± 8.9278 to 49.747 ± 4.4729 MPa and in self-cured acrylic resin from 37.573 ± 10.9237 to 35.569 ± 6.1531 MPa (P < 0.05). Conclusion: The incorporation of aluminum oxide nanoparticles adversely affected acrylic resin flexural strength; however, it increased the thermal conduction.

Evaluation of Alterations in DNA Methylation of CYP3A4 Gene Upstream Regulatory Elements in Gastric Cancer and in Response to Diazinon Treatment.

BACKGROUND: Little is known about cytochrome P450 3A4 (CYP3A4) DNA methylation and transcription alterations in gastric cancer. OBJECTIVE: In this paper, we initially aimed to address the effect of diazinon pesticide on DNA methylation and transcription changes of the CYP3A4 gene in a human gastric cell line. In the next step, we studied the methylation differences of CpG sites within the upstream regulatory regions of the CYP3A4 gene among human gastric cancerous and healthy tissues. METHODS: For the in vitro assay, the methylation changes of the C/EBP response element and transcript level of the CYP3A4 gene were studied following treatment of the AGS cell line with various concentrations of diazinon pesticide. In the next phase, the methylation percentages of 24 CpG sites within or around the upstream regulatory elements, including near promoter, C/EBP binding site, XREM, and CLEM4, in 11 specimens of human gastric cancer tissue were compared to their adjacent healthy tissues. RESULTS: Treatment with 10 µM Diazinon significantly increased the CYP3A4 gene transcription by approximately 27-fold, which was correlated with the hypermethylation of 3 CpGs in C/EBP binding sites, including -5998, -5731 and -5725 (p<0.001 for all comparisons). Results of bisulfite sequencing revealed that the CpG sites which are located in -1521 (p=0.003), -1569 (p=0.027), -10813 (p=0.003), -10851 (p=0.001) and -10895 (p=0.0) bp from transcription start site, were significantly hypermethylated in cancerous tissues comparing to their healthy cohort. CONCLUSION: Hypermethylation of CLEM4 and a region near the core promoter may have a significant association with gastric cancer incidence.

CYP1A1 contiguous hypermethylation within a putative CpG block is associated with breast cancer progression: Feasibility to define boundary motives.

Having broad specificity for xenobiotics metabolism throughout the body, cytochrome P450 (CYP) isoform 1A1 is of key relevance for carcinogenesis. However, the oncogenic potential of its altered transcription and the underlying mechanism has not been well-established in breast cancer. Direct bisulfite sequencing PCR (BSP) of the CYP1A1 promoter, enriched by 113 CpGs within and flanking the xenobiotic response elements (XREs) 2 to 10, in paired cancerous and normal tissues from 40 breast cancer patients revealed three distinctly methylated patterns; unmethylated (XREs 2 to 6) and completely methylated (XREs 7 and 8) CpGs, in common for the normal and cancerous tissues, and a putative 171bp CpG block (XREs 9 and 10) contiguously hypermethylated in the tumor tissues. Increased transcription of CYP1A1, observed for the cancerous tissues, was correlated with the hypermethylation of given CpG block, besides simultaneously being associated with upregulation of the anti-apoptotic BCL-2. Clinical value of the methylation changes, investigated based on the comparisons between the tissue cohorts of different clinicopathological features, exhibited gradual hypermethylation of the corresponding CpG block following disease progression as well as lymphatic involvement. Hypermethylation of given CpG block may has potential to be used as a biomarker for diagnosis and progression of breast cancer.

Dietary patterns and relative expression levels of PPAR-γ, VEGF-A and HIF-1α genes in benign breast diseases: case-control and consecutive case-series designs.

We aimed to study dietary patterns in association with the relative expression levels of PPAR-γ, vascular endothelial growth factor-A (VEGF-A) and hypoxia-inducible factor-1α (HIF-1α) in women with benign breast disease (BBD). The study design was combinative, included a case-series and case-control compartments. Initially, eligible BBD patients (n 77, aged 19-52 years old) were recruited at Nour-Nejat hospital, Tabriz, Iran (2012-2014). A hospital-based group of healthy controls was matched for age (n 231, aged 20-63 years old) and sex. Dietary data were collected using a valid 136-item FFQ. Principal component analysis generated two main components (Kaiser-Meyer-Olkin = 0·684), including a Healthy pattern (whole bread, fruits, vegetables, vegetable oils, legumes, spices, seafood, low-fat meat, skinless poultry, low-fat dairy products, nuts and seeds) and a Western pattern (starchy foods, high-fat meat and poultry, high-fat dairy products, hydrogenated fat, fast food, salt and sweets). High adherence to the Western pattern increased the risk of BBD (ORadj 5·59; 95 % CI 2·06, 15·10; P < 0·01), whereas high intake of the Healthy pattern was associated with a 74 % lower risk of BBD (95 % CI 0·08, 0·81; P < 0·05). In the BBD population, the Western pattern was correlated with over-expression of HIF-1α (radj 0·309, P < 0·05). There were inverse correlations between the Healthy pattern and expressions of PPAR-γ (radj -0·338, P < 0·05), HIF-1α (radj -0·340, P < 0·05) and VEGF-A (radj -0·286, P < 0·05). In conclusion, new findings suggested that the Healthy pattern was associated inversely with the risk of BBD, and this could be correlated with down-regulation of PPAR-γ, VEGF-A and HIF-1α genes, which might hold promise to preclude BBD of malignant pathological transformation.

Association of CYP1A1 M2 (A2455G) Polymorphism with Susceptibility to Breast Cancer in Mazandaran Province, Northern Iran: A Case-control Study.

BACKGROUND: Breast cancer is one of the most frequent women malignancies in the world. The cytochrome P450 1A1 (CYP1A1) is a key enzyme in xenobiotics metabolism. Moreover, CYP1A1 plays a critical role in the etiology of breast cancer by involving in 2-hydroxylation of estrogen. Therefore, single-nucleotide polymorphisms (SNPs) of its coding gene have been verified to be important in cancer susceptibility. The aim of the study was to evaluate the association of CYP1A1 M2 (A2455G) includes rs1048943 of this SNP polymorphism with the risk of breast cancer in Mazandaran province. METHODS: Ninety-six breast cancer patients with known clinicopathological characters and 110 healthy women as control were genotyped for CYP1A1 M2 polymorphisms by the restriction fragment length polymorphism technique. RESULTS: The analysis of CYP1A1 gene (polymorphism M2) showed that the frequency of homozygous wild genotypes (AA), heterozygous (AG), and mutant genotype (GG) in the patient group, respectively, 78%, 22%, and 0%, and also the frequency of genotypes AA, AG, and GG in healthy included 82%, 16%, and 2%, respectively. Statistical analysis by Logistic regression model at P < 0.05 showed no significant correlation between polymorphisms in CYP1A1M2 and breast cancer risk (odds ratio = 0.84, confidence interval = 0.33-2.17). CONCLUSIONS: The results indicated that the M2 allelic genotypes were significantly associated neither with breast cancer risk nor with clinicopathological characteristics in Mazandaran province.

Effect of Ferula persica plant methanol extract on the level of Cox-2 in induced squamous cell carcinoma (SCC) in rat tongue.

Background: More than 90% of oral cancers are cases of squamous cell carcinoma. Standard treatment of cancer includes a combination of surgery, chemotherapy and radiotherapy. Each of these treatments, however, brings about certain problems and side effects. Today herbal medicine, has become a more preferable option in dealing with health problems or preventing them because this type of medicine has better compatibility with the body and does not cause undesirable side effects. In this study , the effect of Ferula persica plant methanol extraction on Cox-2 levels in SCC induced rat tongue is conducted in vivo. Methods: In this lab research, 75 rats from SD race in the age - range of 2/5 - 3 months were selected and put in five groups. In order to induce tongue carcinoma, 4- Nitroquinoline 1 (4 NQO) powder was used 3 times a week for each rat. Furthermore, Ferula persica extract was given to each of the groups in order to examine Cox-2 changes in the blood. Results: Comparison of Cox-2 average in various groups resulted in the observation that there was significant difference between the Cox-2 levels in the groups which had only received carcinogen and the other groups. In this group, Cox-2 level was less and in the group that had received Ferula extract (500 mg) along with carcinogen , Cox-2 level was found to be more than other groups. Conclusion: Ferula persica extract does not have reducing effect on serum Cox-2.

Study of the regulatory promoter polymorphism (-938C>A) of B-cell lymphoma 2 gene in breast cancer patients of Mazandaran province in Northern Iran.

BACKGROUND: The incidence rate of breast cancer has been dramatically increasing since the last decade in Iran, and it is now one of the most common female malignant tumors. B-cell lymphoma 2 (BCL2) family is the most important regulator of apoptosis, and -938C>A single nucleotide polymorphism (SNP) of BCL2 gene promoter has been demonstrated to influence breast cancer susceptibility. In this research, we study the effect of -938C>A allelic variants on breast cancer risk in Mazandaran province at the North of Iran. MATERIALS AND METHODS: This analysis performed on 120 breast cancer patients who underwent surgery in some referenced hospitals at Mazandaran province along with 130 healthy individuals as a control. DNA extracted from peripheral blood samples was applied in polymerase chain reaction-single-strand conformation polymorphism analysis to determine -938C>A genotype. The association of the -938C>A genotype and breast cancer risk as well as clinicopathological characters were analyzed by logistic regression method. RESULTS: Results showed that genotype frequency of AA, AC, and CC genotypes was 10%, 62%, and 28% for case and 28%, 50%, and 22% in control group, respectively. In the logistic regression model, BCL2 - 938C/A variant genotype AA was associated with a decreased risk of breast cancer (P = 0.041) by 0.31-fold (odds ratio = 0.31, confidence interval = 0.091-0.909) compared to CC genotype. However, no significant association found between -938C>A genotype and clinicopathological characters. CONCLUSION: The study showed that AA genotype of BCL2 gene (-938C>A) is associated with decreased susceptibility to breast cancer. Hence, investigating the -938C>A SNP of BCL2 gene promoter could be an appropriate molecular marker to determine individual sensitivity to breast cancer.

Low Expression of the bcl2 Gene in Gastric Adenocarcinomas in Mazandaran Province of Iran.

BACKGROUND: Gastric cancer accounts for about 8% of the total cancer cases and 10% of total cancer deaths worldwide. It is the second lethal cancer after esophageal cancer and is considered the fourth most common cancer in north and northwest Iran. The bcl2 family has a key role in the regulation of apoptosis and change in its expression can contribute to cancer. This study initially scheduled to determine the expression of bcl2 gene in tissue samples of adenocarcinoma cancer patients. MATERIALS AND METHODS: A total of 10 samples of gastric adenocarcinoma and 10 of normal tissues from Sari hospital were selected and after DNA extraction from tissues, bcl2 gene expression assayed by real-time PCR. RESULTS: Our results demonstrated higher expression of the bcl2 gene in control compared with cancer and marginal cancer tissues. CONCLUSIONS: On one hand BCL2 plays an important role as an oncogene to inhibit apoptosis; on the other hand, it can initiate cell cycle arrest at G0 stage. Our observed association between its expression and patient survival is quite conflicting and may be tissue-specific. The data suggest expression both tumoural and non-tumoral(marginal) groups have lowered expression than controls (P>0.05). Due to the low number of samples we could not examine the relationship with clinicopathological features. However, bcl-2 expression may be important for prognostic outcome or a useful target for therapeutic intervention.