Cytotoxic effect of methanolic extracts of Fritillaria imperialis bulbs and Eryngium caucasicum leaves on hepatoma and colon cancer cells

Objective: To evaluate antitumor activities of Fritillaria imperialis and Eryngium caucasicum methanolic extracts on human hepatoma (HepG2) and colon cancer (HCT116) cell lines in comparison to human foreskin fibroblasts as the normal cells. Methods: Methanolic extracts of Fritillaria imperialis and Eryngium caucasicum were prepared by the maceration method. The effect of the extracts at various concentrations (100, 200, 400, 600, and 800 μg/mL) on cell survival was evaluated using the MTT method. Besides, fluorescence staining was used to evaluate death patterns of the cells. Results: MTT assay showed that Fritillaria imperialis significantly decreased the viability of all cell lines after 24 and 48 hours of treatments. However, Eryngium caucasicum extract did not show any significant cytotoxicity effect on the cell lines. Fluorescence staining revealed that Fritillaria imperialis induced apoptosis of HCT116 cells at 550 μg/mL. Conclusions: Fritillaria imperialis extract has antiproliferative and cytotoxic effects on HCT116 and HepG2 cancer cells and therefore, may serve as an anticancer agent.

Inflammation, a key factor in cancer ambush

Inflammatory condition is the consequence of defensive mechanism of immune system against viral and bacterial infection, tissue injury, UV radiation, stress and etc. Persistently acute inflammation leads to chronic phase which is characterized by production of pro-inflammatory mediators from T cells. These molecules [e.g. IL-6, TNF-alpha, IL-1beta and IL-17] are mostly pleiotropic cytokines involved in multiple signaling cascades. NF-kappaB, STAT3, and HIF-1alpha are the major engaged pathways directing to several downstream targets associating with tumorigenesis and inflammation. Carcinogenesis processes such as DNA mutation/damage, proliferation, angiogenesis, apoptosis, and invasion are implicated to inflammation. Clearly there is a closely association between cancer and inflammation reported as "Seven Hallmark of Cancer". The elucidation of relationship between inflammation and cancer and their interaction may result in effective therapy and prevention. Gastric cancer is one of the main cancer involved in complex correlation of inflammation and cancer. Inflammation in gastric epithelium could trigger cellular transformation and promote invasion by inducing immune responses and utilizing signaling cascades. Gastric tumor microenvironment has inverse association by providing cytokines and inflammatory mediators. This closely relationship facilitates gastric tumor development and the induction of chronic inflammation in tumor microenvironment. The current review will focus on describing the possible and critical ways in which inflammation and cancer are linked together with specific view to gastric cancer and inflammation. Finally, it introduces some putative treatment generally used in this way in order to direct more attention for further exploration

Role of Urinary Levels of Endothelin-1, Monocyte Chemotactic Peptide-1, and N-Acetyl Glucosaminidase in Predicting the Severity of Obstruction in Hydronephrotic Neonates

PURPOSE: Antenatal hydronephrosis (AH) is found in 0.5%-1% of neonates. The aim of the study was to assess the urinary concentrations of 3 biomarkers, endothelin-1 (ET-1), monocyte chemotactic peptide-1 (MCP-1), and N-acetyl-glucosaminidase (NAG) in severely hydronephrotic neonates. MATERIALS AND METHODS: Neonates with a history of prenatal hydronephrosis were enrolled in the prospective study in 2 groups. Group 1 included neonates with severe forms of obstruction requiring surgical intervention and group 2 included neonates with milder forms of obstruction without any functional impairment. Fresh voided urinary levels of ET-1, MCP-1, and NAG were measured and their ratios to urinary Cr were calculated. RESULTS: Fourty-two neonates were enrolled into the 2 groups: group 1, 24 patients (21 male, 3 female); group 2, 18 neonates (16 male, 2 female). There were no statistically significant differences between urinary ET-1, NAG, MCP-1 values, and ET-1/Cr and NAG/Cr ratios in groups 1 and 2. The urinary MCP-1/Cr ratio was significantly higher in group 1 than in group 2. For comparison of groups 1 and 2, the cut-off values were measured as 0.5709 ng/mg (sensitivity, 75%; specificity, 67%; positive predictive value [PPV], 71%; negative predictive value [NPV], 71%), 0.927 ng/mg (sensitivity, 77%; specificity, 72%; PPV, 77%; NPV, 72%), and 1.1913 IU/mg (sensitivity, 62%; specificity, 67%; PPV, 68%; NPV, 60%) for ET-1/Cr, MCP-1/Cr, and NAG/Cr ratios, respectively. CONCLUSIONS: The urinary MCP-1/Cr ratio is significantly elevated in neonates with severe obstruction requiring surgical intervention. Based upon these results, urinary MCP-1/Cr may be useful in identification of severe obstructive hydronephrosis in neonates.

Personalized medicine; a bridge between current medicine and the future healthcare

Personalized medicine is a new insight of health care in which interventions is informed by each patient's characteristics; including in genotype and phenotype. Nowadays, rapid advances in genomics, proteomics, and lipomics, which identified individuals differences may lead to produce new drug and diagnostic and screening biomarkers. These allow physicians to determine the type and dosage of drugs based on genetic and genomic characteristics of each patient. Genetic variants that identified by using molecular analysis can be used to screen at risk people on the particular disease, design and administer appropriate medications that are fit to the genetic background, and select the best treatment protocol. Actually, personalized medicine will be the bridge between current medicine and future medicine. In addition to a brief review of history of personalized medicine, this review focused on its required technologies, the role of personalized medicine in pharmacogenetics, prognostic, diagnostic, and target therapy of cancer, neurodegenerative, cardiovascular, allergic and infectious diseases

Resistance to HER2-targeted therapy

Production and approval of trastuzumab [Herceptin[registered sign]] for the treatment of metastatic breast cancer [MBC] was a millstone in antibody-based targeted therapy in the cancer treatment. However, despite the early success in the clinical trials, trastuzumab failed to appreciate the initial attraction due to development of resistance to the drug. Majority of patients who benefit from the drug acquire resistance to it and experience tumor recurrence within 1 year. Several molecular and cellular mechanisms underlying the resistance to trastuzumab have been proposed. In this review, first, we provide a brief history leading to production of trastuzumab. Also we consider the cellular and molecular antitumor effects of trastuzumab and then, we discuss the mechanisms underlying trastuzumab resistance in four levels

Production of cyclin D1 specific siRNAs by double strand processing for gene therapy of of esophageal squamous cell carcinoma

[RNA interference] is a new strategy in gene therapy and biotechnology which provides new viewpoints in treatment of different diseases such as cancer and viral diseases. CCND1 which is a key gene in cell cycle is amplified and over expressed in esophageal cancer. The aim of this study was to produce siRNAs for CCND1, the key gene in cell cycle. dsRNA digestion method was applied by using recombinant human dicer enzyme to cleave in vitro transcribed dsRNA into a pool of 22bp siRNA. Total RNA was extracted and cDNA was produced using RT-PCR. T7 promoter was added to both ends of the DNA template by PCR. RNA was produced from both strands of the DNA using T7 RNA polymerase. After annealing both strands, dsRNA was prepared. Finally siRNA pool was produced by dicer treatment. RNA extraction yield from HN5 cell line was 14.69 micro g/106 cell. The results from beta actin control gene confirmed the cDNA integrity. After optimization, T7 promoter adding was confirmed using gel electrophoresis and DNA sequencing. After optimization dsRNA yield was improved. The best incubation condition was 18h. Each microgram of dsRNA yielded 0.5 micro g siRNA. dsRNA digestion method includes several steps in which the product of each step is used as the precursor for the next step. So optimization and increasing the specificity and product yield should be the most important goals of the study, because the yield of each step has a direct relationship with the final product yield namely; siRNA. Optimizing and increasing the yield, dsRNA digestion method could be a rapid, available and profitable method for siRNA generation, and providing large amounts of siRNA

Prevalence of EPIYA motifs in Helicobacter pylori strains isolated from patients with dyspeptic disorders in northern Iran

Cytotoxin-associated gene A [CagA]-positive strains of Helicobacter pylori are associated with gastroduodenal diseases. Evidences have suggested that the type of H. pylori CagA EPIYA motifs may be associated with recurrent dyspepsia [i.e. gastritis, peptic ulcer, or gastric cancer]. We investigated the prevalence of different EPIYA motifs [A, B, C, or D] in H. pylori strains isolated from patients with recurrent dyspepsia who underwent upper gastrointestinal [GI] endoscopy. We investigated the prevalence of different EPIYA motifs [A, B, C, or D] in H. pylori strains isolated from patients with recurrent dyspepsia who underwent upper gastrointestinal [GI] endoscopy. H. pylori strains were isolated from biopsy specimens of 220 patients with recurrent dyspepsia. The presence of glmM gene, as a housekeeping gene, CagA gene, and pattern of CagA EPIYA motifs were determined using polymerase chain reaction [PCR] method. The association between the type of motifs and disease state was determined by the Chi-square test, Fisher's exact test, and logistic regression. CagA-positive H. pylori strains were identified in 125 [57%] of patients, including 36 [28.6%] gastritis, 31 [24.6%] duodenal ulcer, and 58 [46.4%] gastric cancer. The frequency of pattern of CagA EPIYA motifs were detected as 39 [31.2%] AB motifs, 54 [43.2%] ABC motifs, 32 [25.6%] ABCC motifs,and no D motifs. The risk of gastric cancer occurrence was estimated to be 2.57 times higher in patients infected by strains with ABCC motif when compared with gastritis and duodenal ulcer patients [p=0.03]. Moreover, patients with C-containing motifs were 2.27 times more likely to be afflicted with gastric cancer than with duodenal ulcer. AB motif was more associated with gastritis and duodenal ulcer than ABC and ABCC motifs. The results suggested that CagA-EPIYA ABCC might be associated with gastric cancer, while EPIYA-AB might be associated with duodenal ulcer

Toll like receptor-4 896a/g gene variation, a risk factor for migraine headaches

The pathogenesis of migraine involves immune-mediated mechanisms in the vascular endothelium. Toll like receptor 4 [TLR-4] is a signaling receptor of innate immunity which plays a role in various neuropathologies related to neuron inflammation. This case/control study is aimed to investigate whether TLR- 4 896A/G variation is related to migraine headaches in an Iranian population. A total of 170 migraine patients [130 females, mean age 33.24 +/- 11 years] and 170 age, sex, and ethnicity matched healthy controls [118 females, mean age of 31 +/- 10 years] were recruited. Genotyping was carried out using the tetra primer amplification refractory mutation system [ARMS]-PCR. The frequency of G allele was higher in migraine patients than the controls [15% vs. 4.7%; p<0.0001]. Interestingly, the distribution of heterozygous 896A/G genotype statistically differed between migraineurs and controls [25.3% vs. 8.2%, p=0.00002, OR 3.87, 95% CI; 2.02-7.4]. Multivariate logistic regression analysis indicated that G allele in affected female migraineurs is an independent factor associated with increased risk of migraine [OR 3.2, 95% CI 1.23-8.24, p=0.01]. Our results showed TLR-4 polymorphism as a genetic risk factor for migraine. However, further studies in different populations are required to elucidate the precise role of TLR-4 896A/G mutation in susceptibility to migraine.

Effect of mesenchymal stem cells on doxorubicin-induced fibrosis

The aim of this study was to test the effect of intravenous injection of mesenchymal stem cells [MSCs] on doxorubicin [DOX]-induced fibrosis in the heart. We investigated the mechanisms that possibly mediate this effect. In this experimental study, fibrosis in the myocardium of adult male Wistar rats [weights 180-200 g, 9-10 weeks of age, total n=30] was created by DOX administration. DOX [2.5 mg/kg] was administered intraperitoneally 3 times a week, for a total dose of 15 mg/kg over a period of 2 weeks. MSCs from Wistar rats were separated and cultured in Dulbecco's modified eagle medium [DMEM]. The condition medium [CM] which contained factors secreted by MSCs was also collected from MSCs cultured in serum-free DMEM. Two weeks after the first injection of DOX, MSCs, CM and standard medium [SM] were transplanted via intravenous injection. Four weeks after transplantation, histological [Masson's trichrome staining for fibrosis detection] and molecular [real-time polymerase chain reaction [RT-PCR]] analyses were conducted. In addition, insulin-like growth factor [IGF-1] and hepatocyte growth factor [HGF] in the CM were measured with an enzyme-linked immunosorbent assay [ELISA]. For immunosuppressive treatment, cyclosporine A was given [intraperitoneally, 5 mg/kg/day] starting on the day of surgery until the end of study in all groups. Fibrosis rate and relative gene expression were compared by analysis of variance [ANOVA] and post-Tukey's test. HGF and [IGF-1 in the CM were analyzed by independent sample t test. P<0.01 was considered statistically significant. Our data demonstrated that intravenously transplanted MSCs and CM significantly reduced fibrosis and significantly increased Bcl-2 expression levels in the myocardium compared to the DOX group [p<0.01]. However, there was no significant difference between Bax expression levels in these groups. In addition, secretion of HGF and IGF-1 was detected in the CM [p<0.01]. We conclude that intravenous transplantation of MSCs and CM can attenuate myocardial fibrosis and increase Bcl-2 expression. This may be mediated by paracrine signaling from MSCs via anti-fibrotic and anti-apoptotic factors such as HGF and IGF-1

Effect of fibroblast co-culture on in vitro maturation and fertilization of mouse preantral follicles

The aim of this study was to evaluate fibroblast co-culture in vitro maturation and fertilization of prepubertal mouse preantral follicles. The ovaries of 12-14 day old mice were dissected and 120-150 micro m intact preantral follicles with one or two layers of granulose cells, and round oocytes were cultured individually in alpha-minimal essential medium [alpha-MEM] supplemented with 5% fetal bovine serum [FBS], 100 mIU/ml recombinant follicle stimulating hormone, 1% insulin, transferring, selenium mix, 100 micro g/ml penicillin and 50 micro g/ml streptomycin as base medium for 12 days. A total number of 226 follicules were cultured under two conditions: i] base medium as control group [n=113]; ii] base medium co-cultured with mouse embryonic fibroblast [MEF] [n=113]. Follicular diameters, alone, in addition, to other factors were analyzed by student's t-test and chi-square test, respectively. The co-culture group showed significant differences [p<0.05] in growth rate [days 4, 6 and 8 of the culture period] and survival rate. However, there was no significant difference in antrum formation, ovulation rate and embryonic development of released oocytes. There were significant differences [p<0.05] in the estradiol and progesterone secretion at all days between the co-culture and control groups. Fibroblast co-culture increased survival rate and steroid production of preantral follicles by promoting granulose cell proliferation

Hypersensitivity to house dust mite and cockroach is the most common allergy in North of Iran

Asthma and allergic rhinitis are among the most common diseases in the world. The aim of this study was to detect, by skin prick test, aeroallergens in allergic patients in Sari, Mazandaran in north of Iran. This is a prospective study of skin prick test of aeroallergens in asthma, allergic rhinitis and their combination with clinical diagnosis. Three hundred and seventy five cases aged between 5 to 50 years, were referred to Tooba and Boo-Ali allergic centers of Mazandaran University of Medical Sciences between December 2006 and July 2009. The aeroallergens studied included house dust mites [Dermatophagoides farinae, Dermatophagoides pteronyssinus], cockroaches, feather, aspergillus, Alternaria, pigweed, nettle, oak and maple. Of the studied individuals, 175 cases were males [46.7%] and 200 were females [53.3%], of which 156 [n=41.5%] reacted to allergen extracts. In asthma, allergic rhinitis and their combination, the respective positive percentages were 26.6%, 22.9%, and 32.6% for Dermatophagoides farinae; 26.6%, 25.3%, and 23.3% for Dermatophagoides pteronyssinus; 12.7%, 17.4%, and 11.6% for cockroaches and 16.5%, 4.7%, and 7.0% for the feather. Other allergens were positive up to 5 percent. Total IgE levels were elevated in 56.4%, 53% and 60.5% of asthmatic, allergic rhinitis and the combination group, respectively. Eosinophils count was elevated in 40.5%, 33.2% and 37.2% of the same groups, respectively. The hypersensitivity to house dust mites is very common in north of Iran which may be attributed to the warm and humid weather of this area

Cytokine production in hymenolepis nana infection

Although many experimental studies provide convincing evidence that type II immunity is protective against helminths, recent data in mice demonstrate that Th1 is also important in some cestodes like Hymenolepis nana. Objective: To identify the role of Th1 and Th2 lymphocytes in immunity against H. nana, the levels of IL-12, IFNgamma, IL-5, and IL-13 were determined in serum of humans infected with this cestode. A total of 31 patients [case] with H. nana infection and 30 clinically healthy individuals [control] was included in this study. Measurements of IL-12, IFNgamma, IL-13 and IL-5 in serum samples were performed by solid-phase sandwich enzyme linked immunosorbant assay. Differential leukocyte count was also done. T test, Mann Whitney U test and Wilcoxan W test were used for data analysis. The mean concentrations of IFNgamma, IL-12 and IL-5 in the sera of patients with H. nana infection were higher than the control group, but only the differences between the concentrations of IFNgamma [p<0.001] and IL-13 [p<0.05] in the two groups were significant. There was an increase in the percentage of monocytes, eosinophils and lymphocytes in patients when compared to the controls, but this increase was not significant. Results from the present study in humans are in agreement with experimental studies in animals in which both Th1 and Th2 responses occur in H. nana infection

E-selectin S128R polymorphism leads to severe asthma

The E-selectin mediates the interaction of activated endothelial cells with leukocytes and plays a fundamental role in the pathogenesis of asthma. It has been suggested that an S/R [Serine128 Arginine] polymorphism of E-selectin alters ligand binding function. Our purpose in this study was to determine whether this Serine128 Arginine polymorphism influences the risk of asthma and also to analyze the possible correlation of disease severity in Iranian patients with polymorphism of E-selectin. We studied human E-selectin gene polymorphism in 172 asthmatic patients and 173 healthy volunteers by polymerase chain reaction-restriction fragment length polymorphism [PCR-RFLP]. To determine the severity of the asthma's situation, a questionnaire was prepared requesting the following information: age, sex, clinical signs and symptoms and past medical history. After the participants filled in the questionnaire, all active or ex-smoker patients were excluded. A trained observer assessed airway reversibility, peak flowmetry and spirometry in asthmatic patients. We found increased serum levels of soluble E-selectin [sE-selectin] in asthmatic patients compared with healthy subjects [P <0.0001]. Frequencies of the SS, SR, and RR genotypes were found as 66.3%, 31.4%, and 2.3% in the patients and 91.9%, 8.1%, and 0.0% in control subjects, respectively. The 128 Arg allele was more prevalent in patients than controls [OR 5.78; 95% CI, 3.07-10.86, P<0.0001]. However, in this study the polymorphism was not associated with circulating sE-selectin levels. We found a direct correlation between the level of sE-selectin and the severity of asthma [P=0.001]. On the other hand, there was a close relation between 128 Arginine carriage and disease severity [P<0.0001]. These results suggest that the Ser 128 Arg polymorphism of the E-selectin gene is a genetic factor that may be associated with the severity of asthma