Association of a Genetic Variant in the Cyclin-Dependent Kinase Inhibitor 2B with Risk of Pancreatic Cancer.

Background: Pancreatic cancer (PC) is among the most aggressive tumors with a poor prognosis, indicating the need for the identification of a novel prognostic biomarker for risk stratifications. Recent genome-wide association studies have demonstrated common genetic variants in a region on chromosome 9p21 associated with an increased risk of different malignancies. Methods: In the present study, we explore the possible relationship between genetic variant, rs10811661, and gene expression of CDKN2B in 75 pancreatic cancer patients, and 188 healthy individuals. DNAs were extracted and genotyping and gene expression were performed by TaqMan real-time PCR and RT-PCR, respectively. Logistic regression was used to assess the association between risk and genotypes, while the significant prognostic variables in the univariate analysis were included in multivariate analyses. Results: The patients with PDAC had a higher frequency of a TT genotype for rs10811661 than the control group. Also, PDAC patients with dominant genetic model, (TT + TC), was associated with increased risk of developing PDAC (OR= 14.71, 95% CI [1.96-110.35], p= 0.009). Moreover, patients with CC genotype had a higher expression of CDKN2B, in comparison with TT genotype. Conclusion: Our findings demonstrated that CDKN2A/B was associated with the risk of developing PDAC, supporting further investigations in the larger and multicenter setting to validate the potential value of this gene as an emerging marker for PDAC.

An In Vivo Study of Lactobacillus rhamnosus (PTCC 1637) as a New Therapeutic Candidate in Esophageal Cancer.

Objective: This study is aimed at investigating the effect of probiotic Lactobacillus rhamnosus on esophageal cancer in vivo and in vitro. Methods and Results: In this study, the cytotoxicity effects of L. rhamnosus supernatant and whole-cell culture on a cancer cell line (Kyse30) compared to 5fu were evaluated by the MTT assay. The real-time PCR method was used to analyse the L. rhamnosus supernatant effect on the expression of Wnt signaling pathway genes. An in vivo investigation in nude mice was done to assess the anti-tumor activity of L. rhamnosus supernatant and whole-cell culture. Both supernatant and whole-cell culture of L. rhamnosus reduced cell survival (Kyse30) P < 0.001. The supernatant of this bacterium significantly reduced the expression of Wnt signaling pathway genes. Administration of supernatant and whole-cell culture of L. rhamnosus expressively reduced tumor growth compared to the control group. The effects of this bacterium on tumor necrosis were quite evident, pathologically P < 0.01. Conclusion: This study is the first report that assessed the potential impact of L. rhamnosus, especially its supernatant on esophageal cancer and Wnt signaling pathway genes. Therefore, this bacterium can be a harmless candidate for esophageal cancer therapy.

Signaling pathways associated with structural changes in varicose veins: a case-control study.

INTRODUCTION AND OBJECTIVES: In varicose veins, blood pressure increases in the veins of the lower extremities due to mechanical stimulation and function remodeling. The aim of this study is assessment of Signaling pathways associated with structural changes in varicose veins. MATERIALS AND METHODS: This pilot study was performed on patients with varicose veins, which had undergone surgery. The healthy tissues from trauma patients or vascular bypass without underlying diseases were used for control samples. Hematoxylin-eosin, trichrome, and elastin staining were used for histopathological examination. The levels of MDA (malondialdehyde), total thiol, SOD (Superoxide dismutase) and NO (nitric oxide) level were measured using Elisa kits to evaluate the oxidative stress level. Gene expression levels of MMP2, MMP9, FOXO3a, APOE and p53 genes were determined using Real-time PCR. RESULTS: This study showed, the vascular Vein wall changes are visible in vascular collagen staining. Although these changes are observed in the structure of vascular wall collagen fibers, the accumulation of collagen and elastin was increased in the walls of varicose veins compared to the control group. The amount of nitric oxide and thiol were increased in the varicose group (P < 0.0001). The expression of metalloproteinase2 gene associated with extracellular matrix change was increased in varicose veins. However, the amount of metalloproteinase 9 was decreased in this group compared to control group. FOXO3a, APOE Genes were down-regulated in the varicose veins compared to control group, while p53 gene expression was significantly increased in the varicose group (P < 0.0001). CONCLUSION: This study demonstrated changes in oxidative stress, morphological structure, and aging pathways in varicose when compared to non-varicose veins. The changes in oxidative stress may be associated with the variations in morphological structure and aging pathways which contribute to the pathogenesis of varicose veins.

The effect of redox signaling on extracellular matrix changes in diabetic wounds leading to amputation.

INTRODUCTION: & Objectives: Redox signaling is a critical regulator in the process of wound healing. This signaling pathway can be effective in the development or healing of diabetic ulcers through the ECM.In this study, the structure of extracellular matrix investigated in relation to redox signaling in the tissue of patients with diabetic ulcers that lead to organ amputation. MATERIALS AND METHODS: The case-control design on diabetic patients ulcers as case group and non-diabetic limb ischemia as control were used.Hematoxylin-eosin, trichrome, and elastin staining methods were used for pathological evaluations of ECM. MDA, total thiol, and SOD levels were measured using ELISA kits to assess the oxidative stress level. Also, NO level was measured by using ELISA kits in both groups. Expression levels of genes MMP2, MMP9, and HIF were detected using real-time PCR with SYBR-green assay. RESULTS: The pathological results showed an increase in the thickness of collagen and elastin fibers. Lipids atrophy was visible in the tissue isolated from the diabetic wound group. The amount of MAD to evaluate the level of lipid oxidation in patients with diabetic Ulcer was significantly higher than the control group(p < 0.01). Thiol level was significantly lower in the diabetic ulcer group than in the control group(p < 0.0001). The expression of metalloproteinases 2 and 9 genes in the tissues isolated from diabetic ulcers was lower than the control group(p < 0.0001). While the expression of the HIF gene in this group was higher than the control group(p < 0.0001). CONCLUTION: In the diabetic wound, the HIF secretion due to hypoxic conditions is beneficial for matrix deposition and prevents protease activity, but if the hypoxia persists, it can lead to ECM deposition subsequently increases the tissue pressure, increases of the collagen I-to-collagen III ratio in collagen accumulation that due to more hypoxia , lipidsAtrophy and eventually amputation.

Routine Multiple Duodenal Biopsy during Endoscopy of Dyspeptic Patients Seems Unnecessary for Screening of Celiac Disease.

INTRODUCTION: Celiac disease (CD) is a chronic and common cause of dyspepsia with a rising prevalence worldwide. This study is aimed at investigating the prevalence of CD in dyspeptic patients based on serology and biopsy, determining the associated factors, and assessing the necessity of regular duodenal biopsies from normal mucosa in diagnosis of CD among dyspeptic patients. METHODS: This cross-sectional study was performed on 530 adult dyspeptic patients who underwent gastroduodenoscopy in Imam Reza hospital, Mashhad, during 2016-2018. Demographic characteristics, clinical data, and laboratory analyses were extracted from hospital records. CD was diagnosed based on intestinal biopsy and serum antitissue transglutaminase (anti-TTG) levels. Mucosal lesions were classified according to the modified Marsh classification. Data were analyzed in SPSS with P < 0.05 being considered significant. RESULTS: Overall, 163 males (30.8%) and 367 females (69.2%) with an average age of 46.38 ± 15.54 years were studied. High anti-TTG levels were seen in 36 (6.8%) patients, and duodenal pathologies were seen in 23 (4.5%) patients. Fifteen (2.8%) were diagnosed with CD based on both serology and biopsy. Bloating was the most common type of dyspepsia in CD patients (7, 46.7%), followed by epigastric pain (6, 40%), and postprandial fullness (2, 13.3%). Two CD patients (13.3%) reported a positive family history for CD. Logistic regression model showed that iron deficiency anemia (IDA), anti-TTG level, and Helicobacter pylori infection were predictors of histological changes of CD, whereas IDA was the only independent predictor of CD in dyspeptic patients (OR = 17.65, 95%CI = 1.53-202.52, and P = 0.021). CONCLUSION: CD is prevalent in dyspeptic patients, but routine biopsy from normal-appearing duodenal mucosa is not recommended for all patients. Serological studies, complete history, and careful endoscopic evaluation may provide better cost-effective clinical solutions to improve the diagnostic yield of celiac disease in dyspeptic patients.

Neuroprotective effects of clavulanic acid following permanent bilateral common carotid artery occlusion in rats.

We investigated whether clavulanic acid could improve learning and memory, in rats underwent bilateral occlusion of common carotid artery (2VO). Seventy male Wistar rats were subjected to 2VO, with a 1-week interval between right and left artery occlusions. After 2VO, animals received clavulanic acid (10, 20, 40 mg/kg, intraperitoneally), from day 8 to 20. Spatial memory was assessed in the Morris water maze, 1 week after the induction of 2VO (day 15). The mRNA expression levels of bcl-2, bcl2-associated x protein (bax), caspase-3, inducible nitric oxide synthase (iNOS), and amyloid beta precursor protein (APP) were measured in the neocortex and hippocampus. Clavulanic acid significantly decreased the escape latency and swimming time in the training trial days. As well, it increased time and distance percentage in the target quadrant, while it decreased such factors in the opposite quadrant in the final trial day, compared to 2VO + normal saline animals. Real time-PCR data showed a significant higher mRNA expression of bax, caspase 3, and iNOS in the hippocampus and neocortex of 2VO animal compared to nonoccluded rats. APP increased in the neocortex but not hippocampus. Compared with 2VO animals, clavulanic acid significantly down-regulated the expression of iNOS, caspase 3, and APP, accompanied by diminishing the bax/bcl2 ratio. Our results reveal a potential therapeutic use of clavulanic acid for cognitive dysfunction associated with cerebral hypoperfusion in vascular dementia and Alzheimer disease.

Histological and Serological Epidemiology of Hepatitis Delta Virus Coinfection among Patients with Chronic Active Hepatitis B Virus in Razavi Khorasan Province, Northeastern Iran.

BACKGROUND: Hepatitis delta virus (HDV), as well as hepatitis B virus (HBV), are regarded as one of the main public health issues in developing countries. This retrospective study described histological and serological features of HDV coinfection patients with chronic active HBV in Northeastern Iran. METHODS: The frequency of HDV seropositivity and its impact on serum liver enzyme levels and pathological features were investigated by reviewing clinical and laboratory data. This study contained chronic active HBV-infected patients having admitted the department during 2009 and 2014. RESULTS: The rate of HDV coinfection in chronic active carriers was 21.84%, with a male predominance. HDV seropositive carriers showed significantly higher concentrations of liver enzyme than chronic active HBV monoinfection. Moreover, there was a strong association between degrees of inflammation with HDV-positive patients' enzyme levels. CONCLUSION: The HDV seroprevalence in northeastern Iran was higher than that reported from elsewhere in Iran while comparable to some regions in Middle East, which, in turn, requires more comprehensive tools for diagnosing and screening the blood.

Teratogenic effects of silymarin on mouse fetuses.

OBJECTIVE: Silybum marianum has been used for centuries in herbal medicine for treatment of liver diseases. Currently, there is no data available on the possible effects of silymarin on fetal development. This study aimed to investigate the teratogenic effect of silymarin on BALB/c mice fetuses. MATERIALS AND METHODS: A total of 40 pregnant mice were divided into 4 groups of 10 mice each. Three groups received silymarin at three different doses of 50, 100 and 200 mg/kg/day during gestational days (GDs). The control group received normal saline and tween (solvent). Dams were sacrificed on GD 18 and all fetuses were examined for gross malformations, size and body weight. Malformed fetuses were double stained with alizarin red and alcian blue. RESULTS: Silymarin administration at all doses resulted in reduction of the mean fetal body weights. The abnormalities included limb, vertebral column and craniofacial malformations. Craniofacial malformations were the most common abnormalities, but they were not observed in a dose-dependent manner. The percentage of fetal resorption significantly increased (up to 15%) in all treatment groups. CONCLUSION: Based on our results, silymarin, especially at high doses can lead to fetal resorption, intrauterine growth retardation and limb, vertebral column and craniofacial abnormalities. More precise studies should be conducted about the teratogenic effects of herbal medicine investigating the underlying mechanisms. Thus, caution should be taken when administering S. marianum to pregnant woman.

Neuroprotective and memory enhancing effects of auraptene in a rat model of vascular dementia: Experimental study and histopathological evaluation.

Vascular dementia and Alzheimer disease are most common type of dementia. These diseases have been associated with cognitive decline and affected personal behavioral activities. Moreover, the pattern of cerebral blood flow in mild cognitive disorder has appeared as a predictive indication for the development into Alzheimer's disease. Permanent, bilateral occlusion of the common carotid arteries (2VO) is a standard animal model to study vascular dementia and chronic cerebral hypoperfusion. In present study neuroprotective and memory enhancing effects of auraptene (AUR), a citrus coumarin, were studied in 2VO rats. Different doses (25, 8 & 4mg/kg) of AUR were administered orally. The spatial memory performance was tested with Morris water maze after 2VO induction. Biochemical experiments and histopathological evaluations were also applied to investigate the neuroprotective effect of AUR in brain tissue. In comparison with 2VO group, AUR could significantly decrease the scape latency time in treated rats. Also AUR increased the percentage of time spent and traveled pathway in target quadrant on final trial test day. All behavioral results were confirmed by biochemical and histopathological data. Biochemical data indicated that AUR could decrease malondialdehyde (MDA), as lipid peroxidation indicator, and increase glutathione (GSH) content in cortex and hippocampus tissues. Histopathological data showed that AUR could protect cerebrocortical and hippocampus neurons against ischemia. This study demonstrated the memory enhancing effect and neuroprotective activity of AUR after induction of brain ischemia in a rat model of vascular dementia.

Intratumoral CD68-, CD117-, CD56-, and CD1a-positive immune cells and the survival of Iranian patients with non-metastatic intestinal-type gastric carcinoma.

The purpose of this study was to investigate the prognostic significance of intratumoral immune cell counts in Iranian gastric cancer patients. A historical cohort study was conducted on fifty patients with non-metastatic intestinal-type gastric adenocarcinoma who underwent curative gastrectomy during 2004-2008 in Imam-Reza Hospital, Mashhad University of Medical Sciences. Immunohistochemistry (IHC) staining for CD56, CD68, CD117 and CD1a was performed to detect natural killer cells, macrophages, mast cells and Langerhans cells, respectively. The immune cells were counted, and the patients were then stratified into low or high immune cell counts. The prognostic significance of this grouping was analyzed using the Kaplan Meier method and Cox proportional hazards regression modeling. Thirty eight (76%) male and twelve (24%) female patients were enrolled in the study with a mean (±SD) age of 66.0 (±9.2) years. The median survival time was 15.0 (95%CI: 5.5-24.5) months. Natural killer cells, mast cells and Langerhans cells showed a positive effect on survival, whereas the reverse was true for macrophages. Multivariate analysis showed that the independent prognostic factors were location of the tumor (cardia/non-cardia), stage, the presence of extra-cytoplasmic mucin, tumor associated macrophage status (low/high), and tumor associated Langerhans cell status (low/high). In the studied population, gastric carcinoma proved to have a very poor prognosis. Multivariate analysis showed that the prognostic effect of natural killer and mast cells in tumoral tissue were dependent on the Langerhans cell count, defending the theory that dendritic cells mediate other immunocytes activities.

Immunohistochemistry comparing endoscopic vein harvesting vs. open vein harvesting on saphenous vein endothelium.

OBJECTIVE: The present study attempts to compare the immunohistochemistry (IHC) of von Willebrand factor (vWf) , endothelial cadherin, Caveolin and endothelial Nitric Oxide Synthase (eNOS) in VasoView Endoscopic Vein Harvesting (EVH) versus traditional Open Vein Harvesting (OVH) techniques for Coronary Artery Bypass Graft (CABG) Surgery performed in Javad al Aemeh Hospital of Mashhad, Iran in 2013,. METHODS AND MATERIALS: Forty-seven patients were scheduled for CABG (30 EVH and 17 OVH) among whom patients with relatively same gender and similar age were selected. Three separate two cm vein samples were harvested from each patient's saphenous vein. Each portion was collected from distal, middle and proximal zones of the saphenous vein. The tissues were deparaffinized, and antigen retrieval was done using EZ-retriever followed by an immunohistochemistry evaluation with vWf, e-cadherin, Caveolin and eNOS. In addition, demographic questioner as of Lipid profile, FBS, BMI, and cardiovascular risk factors were collected. Data analyses, including parametric and nonparametric tests were undertaken using the SPSS 16 software. A P value < 0.05 was regarded as statistically significant. RESULTS: The mean age of the EVH and OVH groups were 63.76 ± 9.51 and 63.63 ± 8.31 years respectively with no significant difference between them (p = 0.989). In addition, there was no great difference between the EVH and OVH groups in lipid profile, DM, HTN, smoking history, CVA, and valvular dysfunction (P > 0.05). Qualitative report of vWf, e-cadherin, Caveolin and eNOS reveals no significant difference between the EVH and OVH (P > 0.05). CONCLUSION: This study indicates that VasoView EVH technique causes no endothelial damage in comparison with OVH. This study could be a molecular confirmation for the innocuous of EVH technique.

Serum gastrin 17, pepsinogen I and pepsinogen II in atrophic gastritis patients living in North-East of Iran.

BACKGROUND: Gastric carcinoma is the second most common cause of cancer-related death in Iran. It is well-known that atrophic gastritis is a major risk factor for gastric cancer, which leads to variations in the serum levels of gastrin 17 (G-17), pepsinogen I (P-I), and pepsinogen II (P-II). The aim of this study was to investigate the diagnostic accuracy of these serum biomarkers in the early detection of atrophic gastritis. MATERIALS AND METHODS: A total of 132 dyspeptic patients underwent upper endoscopy and biopsies were taken. The biopsy specimens were evaluated as the gold standard according to operative link for gastritis assessment staging system. Serum levels of G-17, P-I, and P-II were investigated using enzyme-linked immunosorbent assay. Receiver operating characteristic (ROC) analysis was used to calculate the diagnostic indices and optimal cut-off values using Statistical Package for the Social Sciences SPSS statistical software. RESULTS: A total of 67 men and 65 women were analyzed, among which 48 (36.4%) had atrophic gastritis. The mean age was 45.8 (±15.8) years. ROC curve analysis demonstrated that the biomarkers (including pepsinogen I/II [P-I/II] ratio), except for P-I, are diagnostically significant in detecting gastric atrophy. The area under the curve (95% confidence interval [CI]) for G-17, P-I, P-II, and P-I/II ratio were 0.65 (0.55-0.76), 0.42 (0.32-0.53), 0.62 (0.52-0.72), and 0.61 (0.50-0.72), respectively. However, the diagnostic indices were low (sensitivity <50%, specificity < 90%). The prevalence of Helicobacter pylori infection was significantly higher in patients with atrophy against those without atrophy (75.0% vs. 57.4%, P value < 0.0001). CONCLUSION: In the studied population, the serum biomarkers of atrophic gastritis are not useful screening tests due to their low sensitivity.

Comparison of five staging systems of lymph node metastasis in the gastric carcinoma.

BACKGROUND: The presence of metastatic lymph nodes is the most important prognostic factor for gastric carcinoma; however, the optimal system for the accurate staging of lymph node metastasis for patients with gastric cancer remains controversial. This study was designed to compare five systems in relation to the N classification of gastric carcinoma. MATERIALS AND METHODS: This multicentric historical cohort study was conducted on 148 patients with M0 gastric adenocarcinoma who underwent gastrectomy in, five referral hospitals in Iran. Lymph nodes were sectioned, stained with hematoxyl in and eosin. The lymph node status was classified according to the five systems which are: The number of involved lymph nodes (TNM staging), metastatic lymph node ratio (N ratio), and the largest involved lymph node size, largest metastatic nest size and largest metastatic nest to lymph node size ratio. RESULTS: Patients were classified into significant prognostic groups by the five N classification method including the TNM method, N ratio (0, ≤0.15, 0.15-0.4, >0.4), largest involved lymph node size (0, ≤5, 5-11, >11 mm), Largest metastatic nest size (≤1, 1-7.5, >7.5 mm) and largest metastatic nest to lymph node size ratio (≤0.3, 0.3-0.9, >0.9). All of the above systems remained as independently significant prognostic factors in terms of overall and disease free survival time. CONCLUSION: Among the N staging systems we recommend the metastatic lymph node ratio and largest metastatic nest to lymph node size systems, since they are reproducible, simple, have good survival applicability, have prognostic value and include less stage migration especially in patients whom fewer than 15 lymph nodes are dissected.

Primary glomus tumor of the liver.

Primary glomus tumors are extremely rare in the liver. To this date, only two cases of primary glomus tumors of the liver have been reported in the literature. This report is the third case of this specified tumor. The patient was a 50 year-old Iranian woman who presented with a history of epigastric pain and nausea. Abdominal computed tomography (CT) scan showed a huge hepatic mass that replaced the entire left lobe. Surgical biopsy showed that the specimen consisted of a vascular network confined by the proliferation of epithelioid cells with round-to-oval nuclei and eosinophilic cytoplasms. The cells expressed diffuse reactivity for vimentin, smooth muscle actin and CD34, whereas no reactivity was observed with carcinoembryonic antigen, synaptophysin, chromogranin, cytokeratin, and human melanoma black 45. We conclude that glomus tumors must be considered in the differential diagnosis of hepatic masses, with immunohistochemical staining for smooth muscle actin being a useful method to prove the diagnosis.

Pattern of compensatory hypertrophy in contralateral testis after unilateral orchiectomy in immature rabbits.

INTRODUCTION: Our aim was to evaluate effects of hemicastration in immature rabbits on the histology of the contralateral testis after puberty. MATERIALS AND METHODS: Eighteen immature male rabbits were randomly divided into two groups. The first group underwent right or left hemicastration and the second, sham operation. After their puberty, the rabbits underwent the second operation. In the former group the contralateral testis and in the latter, the right or left testis was removed and sent for pathologic examination. The two groups were compared in terms of Leydig cell count, testis volume, and seminiferous tubule count and diameter. RESULTS: The mature rabbits' mean weight at the orchiectomy time, seminiferous tubule count, and seminiferous tubules diameter did not show significant differences between two groups. However, testis volumes and Leydig cell count were significantly higher in the first group with hemicastration prior to puberty. The mean testis volume was 3.24 +/- 2.06 mL in the first group and 1.4689 +/- 0.85701 mL in the second group (P = .03), and the mean Leydig cell count in every 5 microscopic high-power fields was 86.22 +/- 54.96 and 42.00 +/- 18.09, respectively (P = .04). CONCLUSION: Our research demonstrated that prepubertal hemicastration in rabbits led to the compensatory hypertrophy in the contralateral testis after puberty and an increase in the number of the Leydig cells.

Familial lecithin-cholesterol acyltransferase deficiency.

Familial lecithin-cholesterol acyltransferase deficiency is an uncommon autosomal recessive disorder from a heritable defect in esterification of plasma cholesterol. In 1968, the disease was described by Gjone and Norum in Norway. Our case was a 38-year-old woman. Her disease was manifested by presence of lower extremities edema, proteinuria, corneal opacities, increased plasma cholesterol, and hemolytic anemia. Suspicion of the disease was based on renal biopsy, which revealed mesangial expansion and capillary wall widening with clusters of foamy cells in the mesangium. Immunofluorescence study was nonspecific, but specific findings of electron microscopy showed deposition of lipid in the glomerular basement membrane and mesangium. This is the first report of lecithin-cholesterol acyltransferase deficiency in Iran.The diagnosis was confirmed by a low high-density lipoprotein cholesterol concentration, decreased activity of lecithin-cholesterol acyltransferase in plasma, and positive familial history of the disease.