Colorectal Cancer in North-Eastern Iran: a retrospective, comparative study of early-onset and late-onset cases based on data from the Iranian hereditary colorectal cancer registry.

BACKGROUND: The incidence rate of colorectal cancer (CRC) is increasing among patients below 50 years of age. The reason for this is unclear, but could have to do with the fact that indicative variables, such as tumour location, gender preference and genetic preponderance have not been followed up in a consistent mann er. The current study was primarily conducted to improve the hereditary CRC screening programme by assessing the demographic and clinicopathological characteristics of early-onset CRC compared to late-onset CRC in northeast Iran. METHODS: This retrospective study, carried out over a three-year follow-up period (2014-2017), included 562 consecutive CRCs diagnosed in three Mashhad city hospital laboratories in north-eastern Iran. We applied comparative analysis of pathological and hereditary features together with information on the presence of mismatch repair (MMR) gene deficiency with respect to recovery versus mortality. Patients with mutations resulting in absence of the MMR gene MLH1 protein product and normal BRAF status were considered to be at high risk of Lynch syndrome (LS). Analyses using R studio software were performed on early-onset CRC (n = 222) and late-onset CRC (n = 340), corresponding to patients ≤50 years of age and patients > 50 years. RESULTS: From an age-of-onset point of view, the distribution between the genders differed with females showing a higher proportion of early-onset CRC than men (56% vs. 44%), while the late-onset CRC disparity was less pronounced (48% vs. 52%). The mean age of all participants was 55.6 ± 14.8 years, with 40.3 ± 7.3 years for early-onset CRC and 65.1 ± 9.3 years for late-onset CRC. With respect to anatomical tumour location (distal, rectal and proximal), the frequencies were 61, 28 and 11%, respectively, but the variation did not reach statistical significance. However, there was a dramatic difference with regard to the history of CRC in second-degree relatives between two age categories, with much higher numbers of family-related CRCs in the early-onset group. Expression of the MLH1 and PMS2 genes were significantly different between recovered and deceased, while this finding was not observed with regard to the MSH6 and the MSH2 genes. Mortality was significantly higher in those at high risk of LS. CONCLUSION: The variation of demographic, pathological and genetic characteristics between early-onset and late-onset CRC emphasizes the need for a well-defined algorithm to identify high-risk patients.

Prevalence of Mismatch Repair-Deficient Colorectal Adenoma/Polyp in Early-Onset, Advanced Cases: a Cross-Sectional Study Based on Iranian Hereditary Colorectal Cancer Registry.

BACKGROUND: Lynch syndrome (LS) increases the risk of many types of cancer, mainly colorectal cancer (CRC). The purpose of this study was to assess the prevalence of mismatch repair (MMR) deficiency in patients under the age of 50 with advanced adenomatous polyps, aiming at an early diagnosis of LS. METHODS: This retrospective, cross-sectional study included eligible patients with advanced adenomas diagnosed ≤ 50 years of age registered between April 2014 and February 2017 at three pathology centers in Mashhad. Pathological records were reviewed, and colon tissue specimens were analyzed by immunohistochemistry (IHC) staining to identify proteins which serve as markers for LS as they are related to loss of MMR gene (MLH1, MSH2, MSH6, and PMS2) expression. RESULTS: Of 862 consecutive patients, a total of 50 adenomas (54% males, 46% females of mean age 41.24 ± 6.5) met the eligibility criteria. Of the adenomas examined, 20 (40%) had a tubulovillous component, 34 (68%) had high-grade dysplasia, and 30 (60%) had were larger than 10 mm protrusions. None of the patients had loss of MMR protein expression. CONCLUSION: No individual with MMR genetic disorder was identified by IHC screening of early-onset advanced colorectal adenomas. This strategy is therefore not an effective strategy for detecting MMR mutation carriers.

Promoter methylation, transcription, and retrotransposition of LINE-1 in colorectal adenomas and adenocarcinomas.

BACKGROUND: The methylation of the CpG islands of the LINE-1 promoter is a tight control mechanism on the function of mobile elements. However, simultaneous quantification of promoter methylation and transcription of LINE-1 has not been performed in progressive stages of colorectal cancer. In addition, the insertion of mobile elements in the genome of advanced adenoma stage, a precancerous stage before colorectal carcinoma has not been emphasized. In this study, we quantify promoter methylation and transcripts of LINE-1 in three stages of colorectal non-advanced adenoma, advanced adenoma, and adenocarcinoma. In addition, we analyze the insertion of LINE-1, Alu, and SVA elements in the genome of patient tumors with colorectal advanced adenomas. METHODS: LINE-1 hypomethylation status was evaluated by absolute quantitative analysis of methylated alleles (AQAMA) assay. To quantify the level of transcripts for LINE-1, quantitative RT-PCR was performed. To find mobile element insertions, the advanced adenoma tissue samples were subjected to whole genome sequencing and MELT analysis. RESULTS: We found that the LINE-1 promoter methylation in advanced adenoma and adenocarcinoma was significantly lower than that in non-advanced adenomas. Accordingly, the copy number of LINE-1 transcripts in advanced adenoma was significantly higher than that in non-advanced adenomas, and in adenocarcinomas was significantly higher than that in the advanced adenomas. Whole-genome sequencing analysis of colorectal advanced adenomas revealed that at this stage polymorphic insertions of LINE-1, Alu, and SVA comprise approximately 16%, 51%, and 74% of total insertions, respectively. CONCLUSIONS: Our correlative analysis showing a decreased methylation of LINE-1 promoter accompanied by the higher level of LINE-1 transcription, and polymorphic genomic insertions in advanced adenoma, suggests that the early and advanced polyp stages may host very important pathogenic processes concluding to cancer.

Selective capture of plasma cell-free tumor DNA on magnetic beads: a sensitive and versatile tool for liquid biopsy.

PURPOSE: Recently, 'solid tumor biopsies' have been challenged by the emergence of 'liquid biopsies', which are aimed at the isolation and detection of circulating cell-free tumor DNA (ctDNA) in body fluids. Here, we developed and optimized a method for selective capture of ctDNA on magnetic beads (SCC-MAG) for mutation detection in plasma of patients with colorectal cancer (CRC). METHODS: Blood and tissue samples from 28 CRC patients were included for the detection of KRAS mutations. For the tissue samples, mutation analysis was conducted by high resolution melting (HRM) analysis and sequencing. For the SCC-MAG method, ctDNA was isolated from 200 µl plasma from patients with a mutant KRAS gene. For comparison, ctDNA extraction was carried out using a silica membrane-based method, after which mutations were detected using Intplex allele-specific PCR. RESULTS: The mean ctDNA integrity index in plasma samples of cancer patients was 1.03, comparable with that of silica membrane-derived ctDNA (1.011). Notably, the limit of detection for the SCC-MAG approach was lower than that of the silica membrane method and measured 2.25 pg/ml ctDNA in plasma. Our analyses showed that while the silica membrane-based approach was capable of collecting ctDNA from two out of six CRC patient samples (average Cq 34.23), the SCC-MAG captured ctDNA from all samples with an average Cq of 29.76. CONCLUSIONS: We present a robust, reproducible, and highly sensitive method for the analysis of mutation statuses in liquid biopsies. The SCC-MAG method can readily be applied to any nucleic acid target for diagnostic purposes upon careful design of the specific capture probes, and can be multiplexed by several probes to identify multiple targets.

Hepatic Function and Fibrosis Assessment Via 2D-Shear Wave Elastography and Related Biochemical Markers Pre- and Post-Gastric Bypass Surgery.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) exhibits a worldwide distribution and encompasses a wider range of hepatic abnormalities that can culminate in serious clinical outcomes. The growing incidence of NAFLD necessitates more efficient management strategies particularly in clinically severe obese patients. Weight reduction is the cornerstone of NAFLD treatment; therefore, bariatric surgery could be a therapeutic approach in selected obese patients afflicted with NAFLD and other cardiometabolic comorbidities. OBJECTIVE: The present study focused on the potential role of bariatric surgery on hepatic function and NAFLD-related histopathological features measured through a noninvasive method. METHOD: Ninety patients entered to this study and underwent initial preoperative assessments including demographic profile, anthropometric measurements, standard laboratory tests, and hepatic biopsy. Liver stiffness was also evaluated via two-dimensional shear wave elastography (2D-SWE). All assessments were repeated over the subsequent 6 months following surgery except for liver biopsy. RESULTS: Postoperative hepatic elasticity was lessened after 6 months (p = 0/002).The levels of alanine aminotransferase, gamma-glutamyl transferase, total protein, lipid indices, glucose, and platelet count were also improved following surgery (p < 0/001). Further progression of fibrosis was observed in 25% of patients after surgery. CONCLUSION: Bariatric surgery was associated with a favorable impact on anthropometric and hepatic elasticity indices as well as metabolic parameters. The ideal target population for bariatric surgery should be thoroughly addressed, and the underlying risk factors for fibrosis progression need to be controlled before surgery. However, expanded research designed as comprehensive randomized controlled trials are recommended to confirm these findings.

Crosstalk between DNA methylation and gene expression in colorectal cancer, a potential plasma biomarker for tracing this tumor.

Colorectal cancer (CRC), the second leading cause of cancer mortality, constitutes a significant global health burden. An accurate, noninvasive detection method for CRC as complement to colonoscopy could improve the effectiveness of treatment. In the present study, SureSelectXT Methyl-Seq was performed on cancerous and normal colon tissues and CLDN1, INHBA and SLC30A10 were found as candidate methylated genes. MethyLight assay was run on formalin-fixed paraffin-embedded (FFPE) and fresh case and control tissues to validate the methylation of the selected gene. The methylation was significantly different (p-values < 2.2e-16) with a sensitivity of 87.17%; at a specificity cut-off of 100% in FFPE tissues. Methylation studies on fresh tissues, indicated a sensitivity of 82.14% and a specificity cut-off of 92% (p-values = 1.163e-07). The biomarker performance was robust since, normal tissues indicated a significant 22.1-fold over-expression of the selected gene as compared to the corresponding CRC tissues (p-value < 2.2e-16) in the FFPE expression assay. In our plasma pilot study, evaluation of the tissue methylation marker in the circulating cell-free DNA, demonstrated that 9 out of 22 CRC samples and 20 out of 20 normal samples were identified correctly. In summary, there is a clinical feasibility that the offered methylated gene could serve as a candidate biomarker for CRC diagnostic purpose, although further exploration of our candidate gene is warranted.

Expression profile analysis of two antisense lncRNAs to improve prognosis prediction of colorectal adenocarcinoma.

BACKGROUND: Long noncoding RNAs (lncRNAs) are involved in different pathogenesis pathways including cancer pathogenesis. The adenoma-carcinoma pathway in colorectal cancer may involve the aberrant and variable gene expression of regulatory RNAs. This study was conducted to analyse the expression and prognosis prediction ability of two natural antisense transcripts, protein kinase C theta antisense RNA 1 (PRKCQ-AS1), and special AT-rich sequence binding protein 1 antisense RNA 1 (SATB1-AS1) in colorectal low-grade adenoma, advanced adenoma, and adenocarcinomas. METHODS: In this study, from two RNA-seq analyses of CCAT1-ko cells and colorectal carcinoma biopsies having diminished and increased levels of CCAT1 transcription, respectively, we nominated two antisense lncRNAs of PRKCQ-AS1 and SATB1-AS1. Samples from colorectal low-grade adenomas, advanced adenomas, adenocarcinomas, and adjacent tissue were subjected to RT-qPCR to determine the expression of PRKCQ-AS1, SATB1-AS1 along with colon cancer-associated transcript 1 (CCAT1) and cMYC. In addition, we used different bioinformatics analyses and webservers (including GEPIA 2, TCGA, and CancerMine) to elucidate the prognosis prediction value, the expression correlation of sense-antisense pair of genes, and the expression profile of these antisense transcripts at the presence or absence of mutations in the driver genes, or the corresponding sense genes. RESULTS: PRKCQ-AS1 showed a wide range of expression levels in colorectal adenoma, advanced adenoma, and adenocarcinoma. Upregulation of PRKCQ-AS1 was related to a significant decrease in survival of colorectal cancer (CRC) patients. The expression levels of PRKCQ-AS1 and PRKCQ were strong and significantly concordant in normal and cancerous colorectal tissues. While SATB1-AS1 showed a wide range of expression in colorectal adenoma, advanced adenoma, and adenocarcinoma as well, its expression was not related to a decrease in survival of CRC patients. The expression levels of SATB1-AS1 and SATB1 (the sense gene) were not strong in normal colorectal tissues. In addition, where SATB1 gene was mutated, the expression of SATB1-AS1 was significantly downregulated. CONCLUSIONS: We found the expression of PRKCQ-AS1 and SATB1-AS1 at a given stage of CRC very variable, and not all biopsy samples showed the increased expression of these antisense transcripts. PRKCQ-AS1 in contrast to SATB1-AS1 showed a significant prognostic value. Since a significantly concordant expression was observed for SATB1-AS1 and SATB1 in only cancerous, and for PRKCQ-AS1 and PRKCQ in both normal and cancerous colorectal tissues, it can be concluded that common mechanisms may regulate the expression of these sense and antisense genes.

Association Between Gastric Lymphoid Follicles (Precursor Of MALT Lymphomas) And H. pylori Infection At A Referral Hospital In Iran.

PURPOSE: Mucosa-associated lymphoid tissue (MALT) is the most common endoscopic finding in Helicobacter pylori positive patients that can progress to MALT lymphoma after a prolonged antigenic contact. This study was aimed to evaluate the prevalence of lymphoid follicles and aggregates (precursors of MALT lymphomas) in gastric mucosal biopsies and their correlation with H. pylori infection. PATIENTS AND METHODS: In this study, 100 patients who had undergone an upper gastrointestinal endoscopy were enrolled. Five biopsy specimens were taken each patient through screening endoscopy and histopathological changes were evaluated and graded using the Wotherspoon System. The clinical background and H. pylori infection status were also investigated. RESULTS: Among the 100 cases in our series, 79 patients (79%) showed evidence of MALT in at least one biopsy specimen taken from the stomach and 21 cases (21%) had no evidence of MALT. H. pylori infection was detected in 74 (74%) patients. Lymphoid follicles were detected more frequently in H. pylori-positive patients (59%) compared to H. pylori-negative cases (3%) (P<0.001). CONCLUSION: The frequency of lymphoid follicles and aggregates in gastric mucosal is associated with H. pylori infection. Further community-based studies in larger sample sizes using a combination of microscopic methods and PCR assay are required for effective monitoring of H. pylori infection.

Measurement of Liver Stiffness with 2D-Shear Wave Elastography (2D-SWE) in Bariatric Surgery Candidates Reveals Acceptable Diagnostic Yield Compared to Liver Biopsy.

BACKGROUND: Nonalcoholic fatty liver disease (NAFLD) is common among severely obese patients. Two-dimensional shear wave elastography (2D-SWE) has been validated as a noninvasive diagnostic tool for liver stiffness measurement. However, the technical feasibility and accuracy of this method in severely obese patients are still under debate. OBJECTIVE: We aimed to assess the diagnostic accuracy of 2D-SWE in bariatric surgery candidates in comparison with the gold standard liver biopsy. METHODS: Ninety severely obese candidates for bariatric surgery were included. Liver stiffness was measured using 2D-SWE 14 days before liver biopsy. Liver biopsy was taken on the day of surgery. The area under the receiver operating curve (AUROC) was calculated for the staging of liver fibrosis. RESULTS: 2D-SWE was performed in 97.3% of patients successfully. Histologic stages of fibrosis (F0-F4) were detected in 34.2%, 36%, 6.3%, 3.6%, and 0.9% of patients, respectively. The AUROC for 2D-SWE was 0.77 for F1, 0.72 for F2, 0.77 for F3, and 0.70 for F4. In univariate analysis, 2D-SWE values were correlated with BMI, waist circumference, NAFLD activity score (NAS), and steatosis, whereas these components did not affect liver stiffness in multivariate analysis. CONCLUSION: Two-dimensional shear wave elastography of the liver can be feasible and has good accuracy in severely obese candidates for bariatric surgery. Therefore, 2D-SWE may be a good option for assessing liver fibrosis, especially in the early stages of fibrosis to lessen complications of surgery in this population. However, this method should be applied on a larger scale for late stage of fibrosis.

A novel large germ line deletion in adenomatous polyposis coli (APC) gene associated with familial adenomatous polyposis.

BACKGROUND: Familial adenomatous polyposis (FAP) is a familial colorectal cancer predisposition syndrome characterized by the development of numerous colorectal polyps, which is inherited in an autosomal dominant manner. FAP is caused by germ line mutations in adenomatous polyposis coli (APC) gene. Here, we described the identification of a causative APC gene deletion associated with FAP in an Iranian family. METHODS: Diagnosis of FAP was based on clinical findings, family history, and medical records (colonoscopy and histopathological data) after the patients were referred to Reza Radiotherapy and Oncology Center, Iran, for colonoscopy. Blood samples were collected, and genomic DNA was extracted. APC mutation screening was conducted by target next-generation sequencing and quantitative real-time PCR. RESULTS: A novel heterozygous large deletion mutation, c.(135+1_136-1)_(*2113+1_*2114-1) spanning exon 3 to 16 [EX3_16 DEL] of APC gene (GenBank Accession# MG712911), was detected in a proband and all her affected relatives in five generations, which was absent in unaffected family members and normal controls. CONCLUSIONS: This novel deletion is the first report, describing the largest deletion of APC gene. Our novel finding contributes to a more comprehensive database of germ line mutations of APC gene that could be used in medical practice for the molecular diagnosis, risk assessment susceptibility of the disease for the FAP patients.

Prevalence and clinicopathological characteristics of mismatch repair-deficient colorectal carcinoma in early onset cases as compared with late-onset cases: a retrospective cross-sectional study in Northeastern Iran.

OBJECTIVES: Lynch syndrome (LS), a genetically inherited autosomal disorder, increases the incidence of colorectal carcinoma (CRC). We aimed to perform a universal strategy to assess the prevalence and clinicopathological characteristics of early onset CRCs at high risk of LS versus late-onset ones in the Iranian population. SETTING: A local population-based study from Northeastern Iran. PARTICIPANTS: 321 consecutive CRCs and pathology specimen screened between 2013 and 2016. PRIMARY AND SECONDARY OUTCOME MEASURES: Retrospectively, information regarding the clinical criteria was obtained by interviewing the patients with CRC or, their families. Pathologists tested tumours with immunohistochemistry (IHC) staining of four mismatch repair (MMR) proteins (MLH1, MSH2, MSH6 and PMS2). Tumours with absent IHC staining of MLH1 were tested for BRAF mutations to exclude sporadic CRCs. Prevalence of early onset CRCs at high risk of LS and familial CRC type X were assessed as primary and secondary outcome measures, respectively. RESULTS: Of 321 CRCs (13/123 (10.57%), early onset vs 21/198 (10.6%) late-onset) were detected to be MMR-deficient (dMMR). Nine early onset cases and 14 late-onset ones with a loss of MLH1 underwent testing for the BRAF mutation, none of the early onset and four (2.02%) late-onset were recognised as sporadic. The difference in the outcome of IHC-analysis between early and late-onset CRCs at high risk of LS was not statistically significant (p=0.34). Majority of the suspected LS tumours from early onset patients had arisen in distal part (8/11 (72.72%) vs 8/14 (57.14%)), all of which were occurred in the rectum or sigmoid. CONCLUSION: Clinically, these findings suggest that in case of limitation for BRAF testing, the practitioner in Iran may consider managing early onset dMMR cases like LS until access to BRAF testing becomes available to them, before germline testing to accurately diagnose LS.

Which common test should be used to assess spleen autotransplant effect?

BACKGROUND: Historically, total splenectomy was the only choice of treatment for traumatic splenic injuries. However, nonoperative management and spleen-preserving surgical techniques are preferred in modern medicine. In some situations in which the surgeon has to perform splenectomy, spleen autotransplant may preserve the splenic function. Selecting the best method for evaluating the splenic autotransplant effect has been debated for several years. In this study, we compared three common tests in evaluating the implanted spleen function. METHODS: Participants included 10 patients who were candidates for laparotomy and splenectomy. After performing splenectomy, we implanted five pieces of the spleen in the greater omentum of each patient. After 3 months, the implanted spleen function was evaluated by nuclear red blood cell (RBC) scan, serum immunoglobulin (Ig) M level, and presence of Howell-Jolly (HJ) bodies in the peripheral blood smear. RESULTS: All patients had normal peripheral blood smear. The IgM level was lower than normal in one patient, and scintigraphy did not demonstrate the transplanted spleen in another patient. CONCLUSION: All these tests may have comparable results, but because of availability and low cost of peripheral blood smear, which is also easily performed, it can be considered as the first option to evaluate the implanted spleen function.

Screening for Lynch Syndrome in Cases with Colorectal Carcinoma from Mashhad.

INTRODUCTION: Lynch Syndrome (LS) is a genetically inherited autosomal disorder that increases the risk of many types of cancer, especially colorectal cancer (CRC). Identifying these subjects improves morbidity and mortality. We aimed to assess the prevalence of LS with both clinical criteria and universal strategy in Mashhad, Iran. METHODS: In this retrospective study, we screened 322 patients with CRC between 2013 and 2016 in Mashhad, Iran. CRCs were screened based on Amsterdam II criteria, revised Bethesda guideline, and universal strategy. Information regarding the clinical criteria was obtained by interviewing the patients or, their families. Tumors were screened by pathologists with IHC staining of four Mismatch repair (MMR) proteins (MLH1, MSH2, MSH6, and PMS2). Tumors with absent IHC staining of MLH1 were tested for BRAF mutations to exclude sporadic CRCs. RESULTS: Of 322 CRCs, 33 cases were found to be deficient-MMR; 22 of these had concurrent loss of MLH1 and PMS2, followed by concurrent loss of MSH2 and MSH6 in 8 CRCs. Twenty-two cases with a loss of MLH1 underwent testing for the BRAF mutation, 4 of which were recognized as a positive BRAF mutation. Finally, 29 CRCs were found as being positive screen for LS. Poor sensitivity (21.74%) was found for the Amsterdam II criteria and a poor positive predictive value (15.39%) for the revised Bethesda. CONCLUSION: Application of clinical criteria may not be effective enough to identify LS and at least 2-antibody panel (PMS2, MSH6) should be conducted for newly diagnosed CRCs.

EGFR Expression in Patients with Esophageal Squamous Cell Carcinoma and its Association with Pathologic Response to Preoperative Chemoradiotherapy: A Study in Northeastern Iran.

INTRODUCTION: Esophageal squamous cell carcinoma (ESCC) accounts for 80% of all esophageal cancers worldwide. It is the most common histological type of esophageal carcinoma in low-resource countries. ESCC is prevalent in Asian countries, accounting for more than 95% of esophageal cancers. The epidermal growth factor receptor (EGFR) is involved in cancer development, as its gene is often mutated and/or amplified in cancer cells. According to recent statistics, esophageal cancer is the eighth most common cancer in Iran. METHODS: In this retrospective study, we assessed EGFR overexpression, using immunohistochemistry (IHC) in 68 patients with ESCC, undergoing neoadjuvant chemoradiotherapy and esophagectomy in 2011-2014. The treatment protocol included external beam radiotherapy (40 Gy), concomitant with cisplatin 20mg/m2 and 5- fluorouracil (5-FU) 1000 mg/m2 for 4 consecutive days during the first and fourth weeks of treatment. To compare the two groups (EGFR positive and negative) in terms of complete pathologic response, Chi-square test was performed using SPSS version 16. RESULTS: The median age of the patients was 59 years (range: 27-70 years), with a female-to-male ratio of 1.06. Overall, 70% of the subjects showed EGFR overexpression. Complete pathologic response to neoadjuvant treatment was significantly higher in EGFR-positive patients (40% vs. 15.8%, P = 0.05). In all cases, 1- and 3-year overall survival rates were 86.6% ± 4.1 and 48% ± 6.9, respectively. The 1- and 3-year disease free survival rates were calculated as 71.8% ± 5.4 and 44.3% ± 6.5, respectively. The overall survival rate was relatively higher in cases with EGFR overexpression, although the difference was not statistically significant (5-year survival rate: 47.9 ± 8.2 vs. 30.9 ± 13, P = 0.23). CONCLUSION: EGFR overexpression was reported in the majority of patients with ESCC in northeastern Iran. Moreover, EGFR overexpression was significantly associated with complete pathologic response.

Efficacy and toxicity of Samen-ista emulsion on treatment of cutaneous and mucosal bleeding.

Despite new treatment methods, upper gastrointestinal bleeding remains challenging. Samen-ista emulsion is a new agent based on traditional medicine with coagulant properties. The efficacy and safety of Samen-ista were assessed in cutaneous and mucosal bleeding animal models. Coagulant properties of Samen-ista were evaluated using mice tail bleeding assay, marginal ear vein and upper gastrointestinal mucosal bleeding times in rabbits. After 7 days, clinical signs, mortality and end-organ (kidney, liver, lung, brain and gastric mucosa) histopathological changes were also examined. Samen-ista dose-dependently decreased mean cutaneous tail (128 vs. 14 s) and marginal ear vein (396 vs. 84 s) bleeding times. Rabbit's upper gastrointestinal bleeding time was also significantly decreased (214 vs. 15.8 s) upon Samen-ista local endoscopic application. Treatment with Samen-ista for 7 days did not cause any mortality, abnormal signs of bleeding, changes in appetite or significant histopathologicl changes. Samen-ista emulsion is well tolerated and highly effective in achieving hemostasis in cutaneous and mucosal bleeding animal models.

SOX2 expression in gastrointestinal cancers of Iranian patients.

PURPOSE: Gastrointestinal (GI) malignancies are among the 5 most common cancers in Iran, and their high associated mortality rates are attributable to late diagnosis and poor treatment options. SOX2, a transcription factor necessary for maintenance and induction of pluripotency and self-renewal, has been identified as a lineage-survival oncogene in several cancers. In the present study, we examined SOX2 expression in esophageal squamous cell carcinoma (ESCC), gastric adenocarcinoma and colon squamous cell carcinoma (SCC), as well as normal GI tissues, in Iranian patients. METHODS: To elucidate the role of SOX2 in GI carcinogenesis, formalin-fixed tissues were analyzed using immunohistochemistry (IHC), while frozen ESCC samples were studied by quantitative reverse transcription polymerase chain reaction (qRT-PCR). RESULTS: IHC studies indicated presence of SOX2+ cells in a subset of cancerous and normal tissues of stomach and colon, while no significant difference was observed between groups, and no correlation was found between SOX2 expression and tumors grades. Nevertheless, studying ESCC samples with IHC and qRT-PCR revealed overexpression of SOX2 in comparison with normal adjacent tissues. CONCLUSIONS: The present results are in line with other studies and indicate SOX2 up-regulation in ESCC; however, due to our small sample size and contradictory reports, more research is needed to determine the importance of SOX2 in GI cancers.

Hepatitis-C Infection Incidence Among the non-Hodgkin's B-cell Lymphoma Patients in the Northeast of Iran.

BACKGROUND: Various infectious agents like Ebstein Barr Virus (EBV), HTLV-1 and Helicobacter pylori have known as etiologic factors in different sub-types of lymphoma. Although Hepatitis C virus (HCV) has not only been important for its hepatotropism and hepatitis development, but also in recent years its association with some forms of non- Hodgkin's lymphoma (NHL), especially B cell NHL, has reported.In some countries, the rate of B cell NHL development in HCV infected patients was four times more than general population, and then association between HCV infection and B-NHL has proposed in many studies. METHODS: To assess this relationship in our geographic region, in a descriptive study; we have evaluated patients with B-NHL in an oncology center in northeast of Iran for HCV infection. RESULTS: Out of 128 B-NHL patients, HCV Antibody test (with third generation ELISA method) was positive in only one patient, which confirmed with Nested PCR technique. Then the frequency of HCV infection in our patients was 0.7%. CONCLUSION: Respecting to the incidence of HCV infection in general population in Iran, which is between 0.5-1%, we couldn't show higher prevalence of HCV infection in NHL patients than general population, and hence couldn't confirm relation between HCV infection and B-NHL in our region.

The clinical presentation of celiac disease: experiences from northeastern iran.

BACKGROUND This study aimed to explore demographic characteristics and clinical presentations of celiac disease (CD) in Northeastern Iran. METHODS This was a cross-sectional retrospective study of 193 adults with CD who presented to Mashhad University Gastroenterology Clinic between 2008 and 2013. Patient data that included mode of presentation and the presence of any concomitant illnesses were collected. Intestinal biopsy and serum anti-tissue transglutaminase (anti-tTG) were used for diagnosis. Mucosal lesions were classified according to modified Marsh classification. RESULTS Overall, 132 females and 61 males, with a mean age at diagnosis of 32.6 ± 13.2 years were included. The patient's chief complaints in order of decreasing frequency were dyspepsia (24.6%), diarrhea (20%), anemia (12.8%), and flatulence (7.2%). Bone disease was seen (osteopenia, osteoporosis) in 30% of patients. A positive family history of CD was found in 17.9% of cases. There were 64% who had serum anti-tTG >200 units/ml and 78% had a Marsh classification grade 3 on duodenal biopsy. The histology grade (Marsh) did not show any correlation with anti-tTG serum levels, age, body mass index (BMI) or hemoglobin levels. CONCLUSION In Northeastern Iran, CD was seen more commonly in females and with non-diarrheal presentations. Abdominal discomfort, anemia and bone disease were most common primary presentations in this area. Histology grade showed no significant correlation with level of anti-tTG, BMI or hemoglobin levels. We suggest screening for CD in unexplained abdominal discomfort, bone disease and anemia.

Cholestasis and seizure due to lead toxicity: a case report.

INTRODUCTION: Lead poisoning is a major public health risk which may involve major organs. Recently, there have been reports of opioid adulteration with lead in Iran. The following case report is the first of its kind in that intrahepatic cholestasis due to lead toxicity has been described. CASE PRESENTATION: A 65-year-old man presented to the emergency department with abdominal pain, abnormal liver function tests (cholestatic pattern), and normocytic anemia. He had been an opium user for 20 years. Clinical and preclinical findings including the bluish discoloration of periodontal tissues, or Burton's sign, and generalized ileus on abdominal x-ray led us to the possibility of lead poisoning. Lead levels were higher than normal (150 µg/dL). Magnetic resonance cholangiopancreatography (MRCP) and abdominal ultrasound were performed to rule out extra hepatic causes of cholestasis. To evaluate the possibility of lead-induced hepatotoxicity, a liver biopsy was performed. Histological features of lead-induced hepatotoxity have rarely been described in humans. In this patient, focal canalicular cholestasis and mild portal inflammation were confirmed. Thus, treatment with ethylenediaminetetraacetic acid (EDTA) and British anti-lewisite (BAL) were initiated and continued for five days. The patient's liver function tests returned to their normal values, clinical findings including nausea, vomiting, and abdominal pain subsided, and the patient was discharged from the hospital in good condition. CONCLUSIONS: Lead toxicity should always be taken into account in cases of intrahepatic cholestasis with an unknown etiology, especially in a setting where opium abuse is common.

Evaluating stem and cancerous biomarkers in CD15+CD44+ KYSE30 cells.

Digestive system cancers are listed among the ten top causes of cancer-related death worldwide. Cancer stem cells (CSCs) are malignant cells that share some of their characteristics with normal stem cells, including self-renewal and multipotency, and also cancer cells, such as drug resistance and metastasis. Despite many reports on CSCs with digestive system origin, identification and characterization of esophageal CSCs have remained elusive. To examine the validity of routine SC, cancer cell and CSC markers in KYSE30 cells, derived from esophageal carcinoma, cells were first characterized by immunofluorescence and RT-PCR techniques, and then the significance of candidate biomarkers was evaluated in retinoic acid-treated cells by flow cytometry and/or real-time RT-PCR. Meanwhile, to study CD15 (a newly introduced CSC marker) expression in digestive tract cancers, human normal and tumoral tissues of esophagus, stomach, and colon were analyzed by immunohistochemistry. Using several experimental approaches, we show that CD44, but not CD15, could serve as a reliable marker for undifferentiated malignant squamous cells of esophagus. In conclusion, our study confirms the role of CD44 as a CSC marker in KYSE30 cells, an esophageal squamous cell carcinoma cell line, and for the first time indicates the expression of CD15 in non-neural stem-like cancer cells. Although the importance of CD15 was not indicated in diagnosis of digestive cancers, further studies are needed to better understand the biological identity and function of this molecule in non-neural malignancies.