Gastroenterology and Hepatology from Bed to Bench

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Aim: In this study, we aimed to estimate one- to five-year survival rates in Iranian patients with gastric cancer (GC). In addition, we preformed subgroup analyses and meta-regression to explore possible sources of heterogeneity between studies.

 Background: According to literatures, there has been increasing attention to the long-term survival rate in patients with GC in Iran. However, results have been inconsistent and remain controversial in overall survival rates.

Patients and methods: Literature searches were conducted using PubMed, Scopus, and ISI, as well as Magiran, Medlib, SID, and Iran Medex databases. Studies were pooled and summary one to five survival rates were calculated. Univariate and multivariate regression analyses were used to explore possible sources of heterogeneity among studies. Subgroup analyses were also conducted. Analyses were conducted using the STATA statistical software package.

Results: Final analysis of 29361 patients from 26 eligible studies was performed. The overall survival rate (one to five years) in all studies, by meta-analysis of 24, 14, 23, 12 and 22 studies were 52%, 31%, 24%, 22%, and 15%, respectively. Meta-regression analysis showed an increase in one- and five-year survival rate over the time (Reg Coef = 0.016, p= 0.04) and (Reg Coef= 0.021, p= 0.049), respectively. Positive heterogeneity was observed between quality of papers and data sources (P<0.001).

Conclusion: More than half of GC deaths happened in the first year at diagnosis, and another 30% plus they occurred during the second year after confirmed diagnosis. Our results admit lower survival rates in Iran, similar to other developing countries. 

Aim: The present study evaluated the association between G241R and K469E polymorphisms of intercellular adhesion molecule 1 gene and inflammatory bowel disease in Iranian population.

 Background: Inflammatory bowel disease including ulcerative colitis and Crohn’s disease, is a chronic idiopathic inflammatory disease of the gastrointestinal tract. There are two single base polymorphisms of intercellular adhesion molecule 1gene, G241R and K469E, reported to be associated with inflammatory disorders.

Patients and methods: In this case-control study, 156 inflammatory bowel disease patients (110 ulcerative colitis and 46 Crohn’s disease patients) and 131 healthy controls were enrolled. Two polymorphisms of intercellular adhesion molecule 1gene, including G241R and K469E, were assessed by polymerase chain reaction followed by restriction fragment length polymorphism.

Results: The E469 allele of K469E polymorphism was significantly more frequent in Crohn’s disease patients compared to controls (P< 0.05, OR= 1.83; 95% CI: 1.13 to 2.96). The mutant homozygote genotype of K469E polymorphism (E/E) was also significantly more frequent in Crohn’s disease patients compared to controls (P< 0.05, OR= 4.23; 95% CI: 1.42 to 12.59). No difference was observed in the frequency of K469E polymorphism among ulcerative colitis patients compared to controls. There were no significant differences in genotype and allele frequencies of G241R polymorphism among ulcerative colitis and Crohn’s disease patients compared to control subjects.

Conclusion: According to our findings, K469E polymorphism of intercellular adhesion molecule 1 gene may probably participate in the pathogenesis of Crohn’s disease in Iran.

Aim: We aimed to explore the frequency of BRAF V600E mutation in Iranian patients with colorectal cancer (CRC) as well as its association with clinic pathological characteristic of patients.

 Background: CRC is the third leading cause of cancer related death. There is a growing body of data showing the association of BRAF V600E mutation with malignant transformation and clinical outcome of different tumors, including CRC. These findings suggest that BRAF V600E mutation can be used as diagnostic and/or prognostic biomarker for management of cancer patients.

Patients and methods: A total of 85 patients with sporadic tumor were recruited. Braf V600E mutation was investigated using sequencing of extracted DNAs from formalin-fixed paraffin-embedded (FFPE) tumor tissues. Electropherograms were analyzed using Laser-gene 6 software.

Results: More than 95% of patients were in stage I and II and none of them were in stage IV. Patients were mostly below 55 years old and tumors were dominantly located in the distal colon. Of note, no BRAF V600E mutations were detected in our population.

Conclusion: Our results showed no V600E mutation in the BRAF gene in stage I and II of CRC patients. Further studies in multi-center settings are warranted to examine the prognostic and/or predictive value of this marker in different stages of colorectal cancer patients.

Background: Hepatitis A virus (HAV) is highly contagious in individuals living in crowded conditions such as military centers.

Objectives: This study try to investigate the seroprevalence of HAV immunity among Iranian soldiers and determine whether vaccination should be used for military draftees.

Patients and Methods: In this cross-sectional study, a total of 1554 soldiers were recruited through random clustering sampling. Serum anti-HAV antibody was measured by Enzyme-linked immunosorbent assay (ELISA).Statistical analysis was performed using SPSS.

Results: A total of 1554 male soldiers with age ranged from 18 to 34 years (mean age: 21.2±1.9 y) at baseline were evaluated. Overall, 80.3% of the analyzed specimens were anti-HAV seropositive. The seroprevalence rates increased significantly with the age.

Conclusion: Our results suggest that generally vaccination for HAV is not necessary for Iranian military draftees. But the vaccination is recommended for high-risk groups including anti-HAV seronegative soldiers.

Aim: The aim of this study was to evaluate and compare three available methods for mitochondrial isolation from a human cell line to predict the best method for each probable application.

 Background: Organelle isolation is gaining importance in experimental laboratory settings. Mitochondrial dysfunction may affect tumorgenesis process. There are some evidences that transplantation of healthy, intact and active mitochondria into cells containing defective mitochondria may reduce the proliferation. Therefore, isolated mitochondria could be considered as an effective tool for assessment and management of mitochondrial related disorders.

Patients and methods: Mitochondrial isolation from the human liver cell line (HepG2) was performed using two commercially available kits, including Qproteome (Qiagen) and MITOISO2 (Sigma-Aldrich), as well as a manual method. Integrity of inner membrane of mitochondria was assessed by JC-1 staining. Activity of isolated mitochondria was evaluated by DCFH-DA staining, and total yield of isolated mitochondria determined by micro-Lowry method. Finally, relative quantification using Real-time PCR was conducted to compare the mtDNA copy number of mitochondria isolated by three different methods.

Results: Compared to other methods, manual kit resulted in higher yields of total amount of mitochondrial protein and mtDNA copy numbers. Isolated mitochondria by Qproteome kit, showed a higher activity. Finally, the integrity of inner-membrane of isolated mitochondria was significantly higher in Qproteome when compared with the other two methods.

Conclusion: Due to differences in quality, quantity and activity of isolated mitochondria using three techniques discussed here, the method in which best-suited to each research project should be selected according to the distinct features of isolated mitochondria. 

Aim: Evaluation of biological characteristics of 13 identified proteins of patients with cirrhotic liver disease is the main aim of this research.

 Background: In clinical usage, liver biopsy remains the gold standard for diagnosis of hepatic fibrosis. Evaluation and confirmation of liver fibrosis stages and severity of chronic diseases require a precise and noninvasive biomarkers. Since the early detection of cirrhosis is a clinical problem, achieving a sensitive, specific and predictive novel method based on biomarkers is an important task.

Methods: Essential analysis, such as gene ontology (GO) enrichment and protein-protein interactions (PPI) was undergone EXPASy, STRING Database and DAVID Bioinformatics Resources query.

Results: Based on GO analysis, most of proteins are located in the endoplasmic reticulum lumen, intracellular organelle lumen, membrane-enclosed lumen, and extracellular region. The relevant molecular functions are actin binding, metal ion binding, cation binding and ion binding. Cell adhesion, biological adhesion, cellular amino acid derivative, metabolic process and homeostatic process are the related processes. Protein-protein interaction network analysis introduced five proteins (fibroblast growth factor receptor 4, tropomyosin 4, tropomyosin 2 (beta), lectin, Lectin galactoside-binding soluble 3 binding protein and apolipoprotein A-I) as hub and bottleneck proteins.

Conclusion: Our result indicates that regulation of lipid metabolism and cell survival are important biological processes involved in cirrhosis disease. More investigation of above mentioned proteins will provide a better understanding of cirrhosis disease.

Aim: To evaluate the effect of active T. gondii tachyzoites and its products on the gene expression level of IFN-yR1 and IFN-yR2 in a murine model.

 Background: Many studies have shown that the parasite Toxoplasma gondii utilizes different mechanisms to inhibit the function of IFN-?, but the parasite effect on the function of IFN-yR1 and IFN-yR2 is still unclear.

Patients and methods: Toxoplasma lysate product (TLP), excretory/secretory products (ESPs) obtained from cell free and cell culture media as well as active tachyzoites were injected separately to their respective group each consisted of 10 BALB/c mice. One control group of 10 mice received phosphate buffered saline (PBS). All of the mice were euthanized three days after the last injection and then their peritoneal leukocytes were harvested separately. The total RNA was extracted from the samples, converted to cDNA, and the gene expression level of IFN-yR1 and IFN-yR2 was assessed in all of the treated groups relative to the control one.

Results: There was no significant difference between each of the treated groups relative to the control group concerning the gene expression level of IFN-yR2 (P> 0.05). Furthermore, the gene expression level of IFN-yR1 in two groups of TLP (P= 0.04) and ESP obtained from cell free medium (P= 0.008) showed a significant difference relative to the control group.

Conclusion: Findings of this study revealed a new aspect of host-T. gondii interaction in that this parasite is able to downregulate IFN-yR1 to reduce the IFN-y effects on the infected cell. 

Aim: We aimed to study the expression of CD24 and CD133 in colorectal cancer and normal adjacent tissues to assess a relationship between these markers and clinic-pathological characteristics and patient’s survival.

Background: Cancer stem cells are a group of tumor cells that have regeneration and multi-order differentiation capabilities.

Patients and methods: Expression of CD24 and CD133 was studied in a paraffin block of colorectal cancer and normal tissues near tumors with the immuneohistochemical method in patients who were referred to Imam Khomeini Hospital in Sari.

Results: A total of 50 samples (25 males and 25 females) with a mean age of 67.57±13.9 years old with range 28-93 years, included 3 mucinous carcinoma and 47 adenocarcinoma. Expression of CD133 marker was negative in 29 cases and positive in 21 cases. Expression of CD24 in tissue near tumor cells was found in 30% of available samples. The relationship between expressing CD24 with treatment (surgery and chemotherapy) was significant and its relationship with patient’s survival was insignificant statistically. However, there was a clear difference as mean survival age of patients based on CD24 expression was 26.64±18.15 for negative cases and 41.75±28.76 months for positive cases. CD24 and CD133 expressions and their co-expression with other clinic-pathological factors were not significant.

Conclusion: During this study, the relationship between CD24 and treatment type was significant. To confirm this result, various studies with high sample numbers and other stem cell markers are recommended. 

A 62-year-old woman complaining of severe malabsorption was diagnosed with celiac disease based on the findings of flat, small intestinal mucosa and HLA-DQ2 positivity, although celiac serology was negative. This diagnosis was questioned due to the lack of clinical and histological improvement after a long period of strict gluten-free diet. The detection of enterocyte autoantibodies guided to the correct diagnosis of autoimmune enteropathy, leading to a complete recovery of the patient following an appropriate immunosuppressive treatment. Autoimmune enteropathy should be considered in the differential diagnosis of malabsorption with severe villous atrophy, including those cases with negative celiac-related serology.

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