Expression Levels of Vascular Endothelial Growth Factors A and C in Patients with Peptic Ulcers and Gastric Cancer

PURPOSE: Vascular endothelial growth factor (VEGF) is one of the most important growth factors for metastatic tumors. To clarify the role of VEGF-A and C in patients with peptic ulcer disease (PUD) or gastric cancer (GC), we evaluated the expression levels of these two molecules. We also analyzed the effect of Helicobacter pylori infection on VEGF-A and C expression levels. MATERIALS AND METHODS: Patients with dyspepsia who needed diagnostic endoscopy were selected and divided into three groups: non-ulcer dyspepsia (NUD), PUD, and GC, according to their endoscopic and histopathological results. Fifty-two patients with NUD, 50 with PUD, and 38 with GC were enrolled in this study. H. pylori infection was diagnosed by the rapid urease test. After RNA extraction and synthesis of cDNA, the expression levels of VEGF-A and C were determined by quantitative reverse transcriptase polymerase chain reaction. RESULTS: The VEGF-C expression level in the PUD and GC groups was significantly higher than that in the NUD group. Moreover, the VEGF-A expression level in the PUD and GC groups was higher than in the NUD group, although the differences were not statistically significant. Significant positive correlations were also observed between the expression levels of these two molecules in the PUD and GC groups. In addition, the expression levels of these two molecules were higher in H. pylori positive patients with PUD or GC than in H. pylori negative patients of the same groups; however, these differences did not reach statistical significance. CONCLUSIONS: Up-regulation of VEGF-C expression during gastric mucosal inflammation may play a role in the development of peptic ulcers or GC.

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

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

[Evaluation on antibiotic resistance of helicobacter pylori isolated from patients admitted to tooba medical center, Sari]

Helicobacter pylori, which infect approximately one half of the world's population, are an important risk factor in chronic gastritis, peptic ulcer disease, and gastric cancer. H. pylori eradication is now widely recommended as the most effective treatment of peptic ulcer disease. One of the most important reasons for treatment failure is H. pylori resistance to the antimicrobials usage in therapy. The aim of this study was to determine susceptibility patterns of H. pylori isolates in 6 routine anti-microbial agents in Northern Iran. 125 patients from Tooba Medical Center in Sari with endoscopic evidence of dyspepsia complaints were used for obtaining gastric biopsies specimens. Biopsies were sent to the laboratory in thioglycolate broth [transport medium]. Bacteria were primarily cultured on Columbia agar supplemented with 7% horse blood, 7% fetal calf serum. Urease, Catalase and Oxidase activities were used for H. pylori identification. Bacterial suspensions equivalent to 3 Mc. Farlands were spread on plates, along with antibiotic disks and placed in the diameter zone. Inhibition was measured after 3 days of incubation in micro-aerophilic condition. H. pylori were isolated from 116[92.8%] subjects, a total of 125 biopsy specimens. Resistance to metronidazole, amoxicillin, clarithromycin, tetracycline, furazolidone and ciprofloxacin were 71%, 35%, 25%, 9%, 24% and 25%, respectively. Multiple resistance [amoxicillin-clarithromycin-metronidazole] were found in [6]5% of the isolates. Comparison of our data with previous results showed that prevalence of H. pylori resistance to clarithromycin, furazolidone and metronidazole has increased in Iran considerably. Resistance to amoxicillin in our study was too high in comparison with foreign studies. The present study demonstrates the need for continuous monitoring of the antimicrobial susceptibility in H. pylori in order to determine the optimal drug regimens

[Th1 and Th2 immune response in hymenolepis nana infection]

Although many experimental studies provide convincing evidence, that type II immunity is protective against helminthes; recent data in mice reveal that Th1 are also important in some cestods like Hymenolepis nana. To reveal the role of Th1 and Th2 lymphocyte in immunity against H.nana, the level of IL12, IFN, IL5, IL13 was determined in serum of human infected with H.nana. In a case control study in 2006 in Mazandaran Medical Sciences University a total of 31 patients [case] with H.nana infection and 30 clinical healthy people [control] were included in this study. Measurment of IL12, IFN, IL13 and IL5 in serum samples were performed by solid-phase sandwich enzyme linked immunosorbant assay. Differential leukocyte count also was done. T test, mannwhitney test and wilcoxan W test were used for data analysis. The mean concentration of IFN, IL13, IL12, IL5 in the sera of patient with H.nana infection were higher than control group, but only the difference between concentration of IFN [P < 0.05] and IL13 [P < 0.05] in two groups were significant. There was an increase in percentage of monocytes, Eosinophils and lymphocytes in patient when compared to the control group, but this increase was not significant. Results form the present study are in agreement with experimental study in that both Th1 and Th2 responses occurs in H.nana infection

Evaluation of different sperm immunization methods in mice

Background: Antifertility effect of naturally occuring antisperm antibody [ASA] in infertile couples and studies on experimental immunization of various animals with sperm antigens represents ASA as immunocontraceptive target. Despite extensive research on the effects of different factors on sperm immunogenecity and ASA production variable result have been reported

Objective: To study whole sperm immunization in mice

Methods: In an experimental study, whole mice sperm with different adjuvant i.e. complete Freund's adjuvant [CFA], incomplete Freund's adjuvant [ICFA], and cholera toxin subunit-p [CTS-[3] were administrated to mice intramuscularly [IM], subcutaneously [SC], intranasally [IN], intra-peritoneally [IP], intrarectally [IR], intravaginally [IVA] and orally. Control groups were inoculated with phosphate buffer saline [PBS] plus corresponding adjuvant. Immunization was carried out on days 0, 7, 14, 28 and ASA titers were detected by indirect immunoflu-orescence [IFA] technique in sera and vaginal washes of all groups. The IP group was further excluded from the study due to high mortality rate. The results were compared between control and experimental groups by Mann Whitney and Fisher exact tests

Results: The number of positive mice for ASA in IM, SC, IN experimental and control groups were significantly different [P = 0.01, P = 0.01, P = 0.04, respectively]. However, there were no significant differences between IR, IVA, and oral experimental and control groups. No differences were observed between ASA in vaginal washing of all groups. Due to high mortality in IP group it was excluded from the study

Conclusion: It can be concluded that the whole sperm antigen can induce immune response in female mice by IM, SC, IN but not IAV, IR and oral administration routes