Gastroenterology and Hepatology from Bed to Bench

Colorectal cancer is classified in to three forms: sporadic (70-75%), familial (20-25%) and hereditary (5-10%). hereditary colorectal cancer syndromes classified into two different subtypes: polyposis and non polyposis. Familial Adenomatous polyposis (FAP; OMIM #175100) is the most common polyposis syndrome, account for <1% of colorectal cancer incidence and characterized by germline mutations in the Adenomatous polyposis coli (APC, <

Gastric cardia adenocarcinoma (GCA)is one of the few malignancies with unexplained reasons that have increased sharply in developed countries. The purpose of this review was to determine the pathways in GCA to identify new biomarker. So by comparing gene expression in GCA group with normal control identified important pathways. Gene expression data were extracted from the beforehand investigations then differentially expressed genes utilized in DAVID program to explorer and find related pathways. Our findings contain 367 gene names. Out of these 367 proteins, 199 were found to be exclusively expressed in GCA; whereas 168 proteins were detected down-regulated or silenced. The GCA associated diseases based on the differently expressed genes made up of diseases pathway related colorectal cancer, small cell lung cancer, breast cancer and H. pylori infection stomach cancer. KEGG pathways related to GCA contained cell cycle, p53 signaling pathway, DNA replication, toll-like receptor signaling pathway and some other diseases. The GO-discovered categories also demonstrated most biological process and molecular function related to cancer. Up until now, there is no report to introduce influential biomarkers in GCA so, the deregulated genes identified in GCA patterns might be helpful for diagnosis, prognosis and therapies for gastric cancer but validation of these biomarkers is necessary.

Metabonomics and other “omics” fields are essential science in analytical chemistry. Modern analytical instruments such as proton nuclear magnetic resonance (¹H NMR) can be provided the great quantity of analytical information. In order to assign unknown samples, chemometrics methods recognition build classification model based on experimental data.  Firstly some current strategies for regarding disease diagnosis are exhibited in metabonomics studies. Some disease such as crohn’s disease can be difficult to diagnosis since its signs and symptoms may be similar to other medical problems or often mimic other symptoms. Applications of NMR and supervised pattern recognition in the field of metabonomics are also reviewed in recent years. The aim of present study was to review the assessment of ¹HNMR to investigate a metabolic profile of crohn’s disease.

The prevalence of obesity as one of the most health concerns has increased globally. This kind of disease has been accounted for several diseases such as type 2 diabetes, different types of cancer, heart disease, and Alzheimer. Obesity is a multifactorial disease that both environmental factors and genetics play important role in its susceptibly. In molecular biology, characterization of the adipocyte secretome is important in signaling to other organs and in regulating energy balance for evaluating underlines mechanism. Since better understanding of this disease lead to both preventive and post treatment of obesity which is achieved by molecular evaluations, this review underlies the importance of some molecular approaches in the field of obesity.

Aim: This study aimed to determinant association between rs76189946 polymorphism of EGF gene and risk of colorectal cancer in an Iranian population.

Background: Colorectal cancer (CRC) is the third most prevalent cancer in both genders worldwide. The determination of genetic variation becomes a new way to etiology of colorectal cancer. Epidermal growth factor (EGF) is a mitogen that plays an important role in cell growth and tumourigenesis, this protein acts by binding its receptor, EGFR.

Patients and methods: DNA samples taken from totally 125 CRC patients and healthy controls were amplified by polymerase chain reaction (PCR) for the rs76189946 polymorphism. Genotypes were analyzed using restriction fragment length polymorphism (RFLP). Finally to confirm the RFLP procedure, 20 of the PCR products were sequenced using the ABI PRISM 3130xl Genetic Analyzer and chain termination method (Applied Biosystems, Carlsbad, CA).

Results: Genotype distribution and allele frequency was similar in CRC patients and controls individuals. We expect observe C and G allele in both groups but only was found C allele.

Conclusion: In this study for first time we identified genetic distribution of exonic rs76189946 polymorphism in EGF gene both CRC patients and healthy controls. These results suggest there wasn’t association between EGF polymorphism rs76189946 and risk of colorectal cancer in an Iranian population.

Aim: In this study, we was tried to prepare a nano compound with a new way in functionalization as anti gastric cancer candidate.

Background: Functionalization of nanotubes is a useful route for modification of their biologic properties. (3-oxoindolin-2-ylidene) urea is a chemical compound that made of isatin and urea that can be useful in cancer study.

Patients and methods: MWNT-COOH was functionalized by this compound with one-step reaction that is a new class in modification. Product has been investigated by FT-IR, Raman and SEM. Anti cancer investigation with human gastric cells and MTT assay test for measurement of viable cell numbers were also performed.

Results: The two bands at around 2800–2900 cm^-1 which are seen in functionalized product are attributed to the CH stretching of MWNT–COOH defects.

Conclusion: Cellular results demonstrated that the functionalized nano-tube is a more toxic agent compared to other samples for cancer cells and can be used as a candidate material for chemotherapy.

Aim: The propose of this study was to evaluate the probable correlation between exon and intron polymorphisms of p53 gene and their association with clinicopathological aspects of gastritis.

Background: regarding to the decisive role of p53 in the development of a variety of human cancers, a comprehensive study concerning probable correlation between polymorphisms in the p53 intron and exon in gastritis lesions, may open new insight toward gastric cancer development and prevention.

Patients and methods: PCR-Sequencing was done for exons and introns 2-7 on the 97 gastritis and normal samples, age range of 15-83 years. Also, microsatellite status was evaluated using five mono nucleotide repeat markers. Variation at codon 72 was associated with IVS2+38, p53INS3 and IVS3-29. In addition, IVS2+38 had association with polymorphism at codon 36 & 245. Gastritis samples had stable microsatellite except nine patients showing polymorphism for NR-21 and one for Bat-25

Results: Most of patients with stable microsatellites (83.9%) had allele G at codon72 without p53INS3. In addition, all patients with GA and CG at codon 36 / IVS2+38 had stable microsatellites.

Severity and activity of gastritis were in association with genotypes combined of codon 36/IVS2+38 and 245/IVS2+38 respectively. In addition, the profiles of combined variation at codon 72/IVS3-29 and codon 72/IVS6+31 were different between patients with ages less and greater than 45 years.

Conclusion: As, some exon variations of p53 gene specially codon 72, were in association with alterations at introns and their combined genotypes were correlated with microsatellite status, pathological findings and age, therefore, it could be inferred that the these combinations of p53 gene polymorphisms work as a whole, not as single.

Aim: The aim of this study was to evaluate the mortality rates and trends from Gastrointestinal (GI) cancer in Iranian population from 1995 to 2003.

Background: Cancer is the third most common cause of death in Iran. Gastrointestinal cancer is the most important causes of mortality due to cancer. The cancer mortality data is important to monitor the effects of screening program, earlier diagnosis, demographic data and other prognostic factors.

Patients and methods: National death Statistic Reported by the Ministry of Health and Medical Education (MOH&ME) from 1995 to 2003, stratified by age group, sex, and cause of death is included in this analysis. Colorectal cancer (CRC) [ICD-9; 153-154], Gastric cancer (GC) [ICD-9; 151], Pancreas cancer (PC) [ICD-9; 25], Esophageal cancer (EC) [ICD-9; C15] and Hepatocellular carcinoma (HCC) [ICD-9; 20] were expressed as the annual mortality rates/100,000, general and/or per gender, and age group.

Results: The cause specific mortality rate of CRC slightly increased during the years under study and for GC and EC showed a sharp increasing. In contrast, the mortality rate of PC decreased slightly during the years under the study. The rate of HCC mortality moderately increased. All mortality rates were higher for male than female.

Conclusion: Our study indicated remarkable increasing trends in mortality of GI cancer in Iran specifically for CRC and GC. Developing for a GC and EC for both primary prevention and early detection programs and providing the facilities for CRC screening, would be the options to control the mortality and burden of GI cancers in the future.

Aim: The objective of this study was to evaluate the real outcomes of chronic hepatitis C patients, who treated with interferon plus ribavirin (INF-RBV) and peg-interferon plus ribavirin (PEG-RBV).

Background: Despite the PEG-RBV has become a standard treatment of hepatitis C virus (HCV) around the world; and in Iran too, but in developing countries like as Iran, INF-RBV is still used among some patients for treating HCV, due to the high costs of treatment with PEG-RBV.

Patients and methods: The present cross-sectional study was conducted on 77 naïve patients referred to a private gastroenterology clinic between years 2007 through 2009 in Tehran. Patients had participated in this study taking two types of combination therapies, based on standard protocol of the Iranian Ministry of Health. At the end of the treatment, sustain virological response (SVR) rate was evaluated.

Results: The outcomes showed in INF-RBV treatment; 11.6%, 16.3% and 34.9% patients were suffered from relapse, lost follow-up their treatment and non-responder, respectively, and finally 37.2% of the patients reached SVR. In PEG-RBV treatment outcomes were as follows; 2.9%, 14.7% and 14.7% patients were non-responder, lost follow-up their treatment and suffered from a relapse, respectively, and 67.6% of the patients reached SVR. The multivariate-adjusted odds ratios of outcomes showed that treated with PEG-RIB and also genotype 3a than the others genotypes in this treated had more chance to achieved SVR.

Conclusion: The findings of the present study showed that the rate of SVR in patients who treated with PEG-RBV significantly was higher than patients who treated with INF-RBV. Also in PEG-RBV the chance of achieving SVR is higher among the patients with genotype 3a than among those with other genotypes.

Aim: This study aims to employ random effect model to evaluate prognostic factors of hepatitis C.

Background: In recent years, Hepatitis C virus (HCV) infection has been a major cause of liver diseases worldwide and represents a major public health problem. Evaluation of risk factors and a community intervention in order to decrease the problem is one of the solutions which help protect people from the infection.

Patients and methods: The data was collected from a longitudinal study during 2005-2010. The response variable in this study was the viral load of each HCV patient during the treatment, immediately after the treatment and 3 to 4 months after the end of the treatment. The outcome variable of interest is the viral load of HCV patients. For analyzing repeated measure viral load of HCV patients, random effect models were used.

Results: The results obtained from random effect model showed that treatment protocol and time statistically significant. The variance component was statistically differing with zero.

Conclusion: According to the results time had a positive effect on rate of viral load of patient. Combination therapy of Peg-interferon plus Ribavirin increased the rate of virological response.

Aim: The aim of this study was to investigate the prevalence of GBV-C among Iranian HBV positive patients using PCR-RFLP technique.

Background: GBV-C was a member of flaviviridae family and recently propose to classify as members of a fourth genus in this family, named Pegivirus  and suggest that at least one quarter of the world's population has been infected with this virus. GBV-C can be transmitted via the blood-borne route, although vertical and sexual transmission is very well documented.

Patients and methods: 100 serum samples were collected from HBsAg positive patients in 2011-2012. RNA was extracted with Qiagene mini kit. cDNA was synthesized by Reverse Transcriptase method and amplified by Semi- nested PCR method .After designing specific primers, the semi nested PCR was optimized, then  sequences of PCR products were analyzed with software such as neb cutter, and sites of restriction enzymes were determined and suitable enzymes were selected for RFLP (Restriction Fragment Length Polymorphism).

Results: PCR products were analyzed in 2% agarose gels containing ethidium bromide and were visualized with ultraviolet (UV) light. A 230 bp band was observed in comparison with 100 kb ladder; this band indicates our target gene of GBV-C genome have been isolate from serum samples.

Conclusion: It seems that Co-infection of GBV-C and HBV are common and This method had acceptable sensitivity for detecting GBV-C and determining its genotype, and more affordable than the other techniques; so the results of this study showed the prevalence of GBV-C were 12 serums of 100 serums HBsAg positive in goal population and one sample from 12 GBV-C positive serums was genotype 3 and the others were genotype 2.

Aim: We aimed to investigate the prevalence of Human Bocavirus isolates among Iranian patients suffering from acute gastroenteritis.

Background: Human Bocavirus is a new parvovirus that has been identified in association with gastroenteritis. Limited data are available about HBoV in Iran.

Patients and methods: Viral DNA was extracted from all 294 stool samples. HBoV DNA was detected in extracted samples by polymerase chain reaction (PCR) amplification of a 354 bp of noncapsid protein 1 (NP1) gene. In addition, all samples were also subjected to a nested PCR to amplify a 455 bp of nonstructural 1 (NS1) gene.

Results: The main clinical symptoms among HBoV positive patients were diarrhea (77.7%), fever (62.9%), vomiting (55.5%), and anorexia (59.2%). NP1 PCR was positive in 8 samples (2.72%), NS1 was positive in 16 patients (5.44%) and 3 samples had positive results in both regions (1.02%).

Aim: The aim of this study was to search for metabolic biomarkers of Crohn’s disease (CD).

Background: Crohn's disease (CD) is a type of inflammatory bowel disease that causing a wide variety of symptoms. CD can influence any part of the gastrointestinal tract from mouth to anus. CD is not easily diagnosed because monitoring tools are currently insufficient. Thus, the discovery of proper methods is needed for early diagnosis of CD.

Patients and methods: We utilized metabolic profiling using proton nuclear magnetic resonance spectroscopy (¹HNMR) to find the metabolites in serum. Classification of CD and healthy subject was done using partial least squares discriminant analysis (PLS-DA).

Results: According to PLS-DA model, we concluded that just using one descriptor CD and control groups could be classified separately. The level of lipid in blood serum of CD compared to healthy cohorts was decreased. For the external test set, the classification model showed a 94% correct classification of CD and healthy subject.

Conclusion: The result of classification model presents that NMR based metabonomics is key tool as well as insight into potential targets for disease therapy and prevention.

Aim: The purpose of this study was to determine quality of life in peptic ulcer patients referring to Al-Zahra hospital of Isfahan.

Background: Peptic ulcer disease (PUD) is one of the most prevalent diseases. Its prevalence is 6-15% and about 10% of people experience its symptom in their life. PUD can have a considerable impact on patients’ quality of life (QOL).

Patients and methods: This descriptive- analytic survey was done on 93 randomly patients referred to Al-Zahra hospital of Isfahan city in Iran. Data gathering was done via questionnaire including five domains: physical, psychological, social, behavioral and economical. For data analysis, t-test, Pearson correlation and ANOVA test were used.

Results: 93 patients with mean age of 38.54 years, including 43 (46.2%) women and 54 (53.8%) men, were studied. There was a negative significant between quality of life and age and between disease duration and psychological, economical domains and between the mean of QOL scores in physical and social domains with the number of cigarette per day, also there was significant relation between social domain and gender, and physical, psychological and behavioral domains with marital status; Physical, social domains with smoking. Also there was a negative significant between physical, social and behavioral domains with years of smoking.

Conclusion: Study results showed that quality of life is in a relatively good level among patients, thus some diseases such as peptic ulcer can effect on quality of life. So, treatment and prevention of these diseases may improve their quality of life.

Keywords: Quality of life, Peptic ulcer, Patients.

Aim: This study was conducted to evaluate the influence H. pylori infection and anti-CagA status on the efficacy of Omeperazole 20 m.g. b.d. for patients with endoscopic oesophagitis.

Background: The influence of Helicobacter pylori (H. pylori) infection and its virulent strain (cytotoxin-associated gene A: CagA) has not been evaluated on efficacy of treatment for patients with erosive oesophagitis in Iran.

Patients and methods: One hundred and ten patients (55 H. Pylori positive and 55 H. Pylori negative) with endoscopic evidence of oeosphagitis were enrolled in this interventional study and treated with Omeprazole 20 m.g. b.d. Healing was assessed at repeat endoscopy after 8 weeks of treatment. H. Pylori infection and anti-CagA-IgG (immunoglobulin G) antibodies were determined for each subject by the rapid urease test, pathological assessment and ELISA.

Results: At repeat endoscopy, following 8 weeks of Omeprazole 20 m.g. b.d. therapy, endoscopic healing of oesophagitis had occurred  in 32 % of the HP +ve patients and 23 % of the HP –ve patients (chi square p<0.01). Among the HP +ve endoscopic healing occurred resolved in 11 (32.4 %) of the CagA +ve patients and 19 (90.5 %) of the CagA –ve patients. This difference was significant (chi-square p <0.001).

Conclusion: H. pylori infection and the CagA virulence factor are associated with an increased rate of healing amongst patients with endoscopic oesophagitis treated with Omperazole 20 m.g. b.d. compared to patients without H. pylori infection.

Purpose: Evaluating the efficacy and safety of adding the probiotic Saccharomyces boulardiito standard triple therapy for eradication of Helicobacter pylori.

Methods and materials: In this randomized controlled trial we evaluated 160 adult patients with biopsy confirmed H.Pylori infection referred to gastroenterology ward of Taleghani hospital. The patients were randomized into two treatment regimens: patients in group A (n=80) were given amoxicillin (1000 mg, b.i.d), clarithromycin (500 mg, b.i.d), omeprazole (20 mg, b.i.d) and probiotic of saccaromyces boularidi (Yomogi) (250 mg, b.i.d) for 14 days, moreover patients in group B (n=80) were given amoxicillin (1000 mg, b.i.d), clarithromycin (500 mg b.i.d) and omeprazole (20 mg,b.i.d) for 14 days.

Results: 160 patients (66 male 41.25%, 94female 58.75%) with the mean age of 47.1±11.4 years were evaluated. The success rate for H. pylori eradication in group A was higher 75(87.5%) than group B 65 (81.2%), but the difference between two groups was not significant (p=0.350). Moreover, in case group side effects as nausea, diarrhea, abdominal discomfort and bloating were significantly lower than control group in first and second weeks.

Conclusion: The result of this study showed that saccaromyces boularidi decreased the adverse effects associated with H.pylori therapy but did not significantly decrease the eradication rate of H.pylori.

Aim: The aim of this study was to determine the concentration of salivary ?₂M as a marker of viral proliferation in HBS Ag⁺, HBV DNA PCR⁺ and Hbs Ag⁺ and HBV DNA PCR^- subjects.

Background: Beta-2 microglobulin (?₂M) is responsible for transmission of viral antigens such as Hepatitis B (HBV) on the surface of liver cells as part of an HLA complex.

Patients and methods: In this case control study, 25 PCR⁺ and 2 PCR^- patients were included. 5 mL of the saliva sample was obtained from all patients and salivary ?₂M level was measured using nephelometer. The data was evaluated by the descriptive, chi square and t tests.

Results: 72% of the PCR⁺ patients received medications and in contrast, 85.7% of the patients with PCR^- did not take any medication (P<0.001). The average salivary concentration ofBeta-2 microglobulin in the PCR⁺ group (5.28 ± 5.45 mg/deciliter) was more than PCR^- group (1.51±0.77) and this difference was statistically significant (P=0.003).

Conclusion: the salivary B₂Mlevel can be used as a marker of viral proliferation in patients with hepatitis B.

Aim: The aim of this study was to evaluate the effect of an aerobic exercise on serum level of liver enzymes and liver echogenicity in patients with non alcoholic fatty liver disease.

Background: Non alcoholic fatty liver disease (NAFLD) has different prevalence in various parts of the world. Obesity and low physical activity are main risk factors for the development of NAFLD.

Patients and methods: Ninety patients diagnosed by ultrasound as NAFLD was evaluated in a clinical trial as case (A) and control groups (B). The effect of aerobic exercise on changing in liver enzymes and liver echogenicity was assessed in cases compared with control group which were only on medical therapy.

Results: The mean age in the sample group was 37.6±8.3 in the 17-56 yrs age range. In group A, fatty liver was in stage I in 26 (57.8%) cases, stage II in 17 (37.8%), and stage III in 2 (4.4%). In group B, fatty liver was in stage I in 30 (66.7%) patients, in stage II in 14 (31.1%) and stage III in 1 (2.2%). After an aerobic exercise, serum level of liver enzymes and liver echogenicity in patients with non alcoholic fatty liver disease was significant improved in case group (ALT (P= 0.0001), AST (P=0.01).

Conclusion: Considering the present study, we can suggest that a controlled aerobic exercise schedule can be helpful in the association of medical therapy in the treatment of NAFLD.

Aim: This study was designed to investigate about findings of Impedance pH-monitoring in patients with atypical symptoms of GERD.

Background: The routine treatment with proton pump inhibitors is not very effective in patients with atypical symptoms of Gastro-Esophageal Reflux Disease (GERD). It may be due to non-acid reflux which cannot be monitored using common methods such as pH-metry. Therefore, in these patients combinational procedure of multi channel intraluminal impedance and pH monitoring has been suggested as a new approach.

Patients and methods: sixty patients with atypical symptoms of GERD underwent combined impedance pH-monitoring. pH of reflux episodes, type of material refluxant, blous cleaning time, proximal extension and symptoms association probability (SAP) was recorded for each patient.

Results

Aim: The purpose of this study is to analyze the cost of GERD and functional dyspepsia for investing its related factors.

Background: Gastro-oesophageal reflux disease GERD and dyspepsia are the most common symptoms of gastrointestinal disorders. Recent studies showed high prevalence and variety of clinical presentation of these two symptoms imposed enormous economic burden to the society.

Cost data that related to economics burden have specific characteristics. So this kind of data needs to specific models. Poisson regression (PR) and negative binomial regression (NB) are the models that were used for analyzing cost data in this paper.

Patients and methods: This study designed as a cross-sectional household survey from May 2006 to December 2007 on a random sample of individual in the Tehran province, Iran to find the prevalence of gastrointestinal symptoms and disorders and its related factors. The Cost in each item was counted. PR and NB were carried out to the data respectively. Likelihood ratio test was performed for comparison between models. Also Log likelihood, Akaike Information Criterion (AIC) and Bayesian Information Criterion (BIC) were used to compare performance of the models.

Results: According to Likelihood ratio test and all three criterions that we used to compare performance of the models, NB was the best model for analyzing this cost data. Sex, age and insurance statues were being significant.

Conclusion: PR and NB models were carried out for this data and according the results improved fit of the NB model over PR, it clearly indicates that over-dispersion is involved due to unobserved heterogeneity and/or clustering. NB model in cost data more appropriate fit than PR.