Association of MicroRNA Polymorphisms With Hepatocellular Carcinoma in an Iranian Population

BACKGROUND: Single nucleotide polymorphisms (SNPs) can modulate various biological processes by influencing microRNA (miRNA) biogenesis and altering target selection. Common SNPs may alter the processing of miRNA and may be associated with hepatocellular carcinoma (HCC). We investigated the relationship between miR-499A>G, miR-149C>T, miR-196a2T>C, and miR-146aG>C and HCC susceptibility, examining the interaction of the miRNAs with hepatitis B virus (HBV). METHODS: We evaluated the associations of miR-499A>G (rs3746444), miR-149C>T (rs2292832), miR-196a2T>C (rs11614913), and miR-146aG>C (rs2910164) with HCC susceptibility in 100 HCC patients (70 males and 30 females) and 120 healthy controls (70 males and 50 females), using the PCR-restriction fragment length polymorphism method. RESULTS: For miR-499A>G, the frequencies of the AG genotype and G allele were higher in female HCC patients than in female controls (P=0.02 and 0.045, respectively). The frequency of the A allele was higher in HBV-positive HCC patients than in controls (P=0.019). For miR-149C>T, the frequency of the CC genotype was higher in female HCC patients than in female controls (P=0.009). For miR-196a2T>C, the frequencies of the CT and CC genotypes and the C allele were higher in HBV-positive HCC patients than in controls (P C polymorphisms did not differ between HCC patients and controls. CONCLUSIONS: miR-499A>G, miR-149C>T, and miR-196a2T>C were associated with the development of HCC in women and/or that of HBV-related HCC. They can be considered genetic risk factors for the development of HCC among Iranians.

Frequency of NRAS gene mutation in wild type KRAS and BRAF colorectal cancers; a single center study

Background: Incidence of colorectal cancer is increasing in countries such as Iran. Molecular biomarkers play very important role in the diagnosis, treatment, and prognosis of this cancer. Mutation in the RAS family [including KRAS and NRAS] is one of these important molecular biomarkers, which should be tested before starting treatment with anti-EGRF [Epidermal growth factor] drugs

Objectives: There has been very few reports about the frequency of NRAS mutation from Iran and no study from south of the country. In this article we will describe our experience about the frequency of NRAS mutation in colorectal cancers from the largest referral center in the south of Iran

Methods: During 5 years [2011-2015], we had 52 cases of colorectal cancers with wild type KRAS and BRAF in the hospitals affiliated to Shiraz University of Medical Sciences with enough tissue for molecular studies. NRAS mutation analysis was performed on paraffin embedded formalin fixed tissue of these cases by polymerase chain reaction [PCR]-sequencing method

Results: Among these 52 cases of colorectal cancer with wild type KRAS and BRAF, there has been 3 [5.7%] cases with mutant NRAS. One of the mutations has been in codon 12 and two in codon 61. No mutation in codon 13 was found. All the three cases were women with stage IV and well differentiated histomorphology

Conclusion: Our results showed that frequency of NRAS mutation in colorectal cancer is rare, which is very close to other studies from different geographic areas of the world

Primary squamous cell carcinoma of the liver: a case report

Primary squamous cell carcinoma [SCC] of the liver is a very rare entity and to the best of our knowledge only 35 cases have been so far reported worldwide. Squamous cell carcinoma is usually diagnosed in skin, head and neck, respiratory tract, esophagus, cervix, and rectum. However, it can rarely occur in the liver as a primary tumor. It has been reported to be associated with liver cysts, chronic inflammation of biliary ducts, and biliary stones. Primary SCC of the liver has an aggressive behavior and poor prognosis. Herein, we reported a middle-aged lady presented with a hepatic mass. Biopsy of the mass revealed SCC and no primary source, other than the liver itself, could be found despite extensive examinations. That is why the tumor was considered to be a primary occurrence of SCC in the liver

Correlation between serum ferritin level and histopathological disease severity in non-alcoholic fatty liver disease

Background: Non-alcoholic fatty liver disease [NAFLD] is one of the most common causes of chronic liver disease worldwide. Recently several parameters, such as serum ferritin, have emerged as possible predictors for the severity of NAFLD and insulin sensitivity. We aimed to investigate the value of serum ferritin level as a useful biomarker for the prediction of histopathological disease severity in non-alcoholic steatohepatitis [NASH], the necro-inflammatory form of NAFLD

Methods: This was a prospective cross sectional study in which demographic, clinical, histological, laboratory, and anthropometric data of 30 adult patients with biopsy-proven NAFLD/NASH were analyzed

Results: In our patients population with mean age of 37.9 years and mean BMI of 26.5, statistical analysis did not show a significant difference between the three grades of steatosis in the mean ferritin levels [p=0.559]. It was also observed that ferritin level did not have a significant correlation with the stage of fibrosis [p =0.228]. The mean transferrin saturation did not show significant difference in different stages and grades of NASH [p =0.260 and 0.944, respectively], either

Conclusion: Serum ferritin level may not be useful as a single marker for the prediction of histopathological severity of disease in young patients with NASH who are not morbidly obese

False negative and false positive rates in common bile duct brushing cytology, a single center experience

Aim: In this study we tried to find out the accuracy of biliary tract brushing cytology in our center as the largest referral center in the south of Iran

Background: Common bile duct brushing cytology has been introduced as the method of choice for the diagnosis of pancreaticobiliary malignancies. However, there have been controversial reports about the sensitivity, specificity and overall accuracy of this method in the English literature

Methods: During the study period [2012-2016] there has been 166 cases of common bile duct brushing cytology taken during endoscopic retrograde cholangiopancreatography [ERCP]. One case has been excluded because of inadequate number of cells in the cytology smear. All the smears have been stained by routine cytologic stains and screened by cytotechnologists and diagnosed by expert cytopathologist. Final diagnosis by biopsy has been considered as the gold standard

Results: According to the final histologic diagnosis as the gold standard, there were 22 false negative and 7 false positive cases. All of the false positive cases have been suspected cases in the background of primary sclerosing cholangitis. The most common final diagnosis of false negative cytologic diagnoses has been intrahepatic cholangiocarcinoma in which no malignant cell has been identified in the presence of adequate number of normal ductal epithelial cells

Conclusion: Common bile duct brushing cytology is the method of choice for the diagnosis of pancreaticobiliary tract malignancies; however, having high specificity [90%], the sensitivity is low [56%]. Cytologic diagnosis of biliary tract malignancies should be made with caution in the patients with primary sclerosing cholangitis. Also it is important to know that high false negative rate is present in common bile duct brushing cytology especially in the cases of intrahepatic cholangiocarcinoma without extension into extrahepatic ducts

Incidental gall bladder adenocarcinoma in cholecystectomy specimens; a single center experience and review of the literature

Background: Gallbladder adenocarcinoma is the most common malignant tumor of the biliary tract. Most of gall bladder cancers are detected incidentally only after pathological examination of the surgical specimens. In this study we investigated the characteristics of incidental gallbladder cancers in our center and also reviewed the current literature regarding the diagnosis and treatment of such incidentally detected tumors in the gall bladder

Methods: We retrospectively reviewed all of the cholecystectomy specimens in the archives of Pathology Department in the hospitals affiliated to Shiraz University of Medical Sciences in the study period [2010-2016]. Clinicopathological characteristics were extracted from the patients' clinical charts, which included symptoms, radiological findings, laboratory data, and surgical procedures as well as outcome

Results: During these 7 years we identified 18 cases of incidental gall bladder cancer, consisted of 13 women and 5 men with the age range of 32 to 85 [62.5 +/- 14.2] years detected after pathological study of the resected gall bladders among more than 4800 resected gall bladders. During the period, only two patients were operated on with the impression of gall bladder adenocarcinoma, which was not included in the study. Ten cases were T1 and eight were T2 at the time of surgery. They have been followed up for 1-7 years, during which, six cases of T2 died. All of the T1 cases are alive and symptom free

Conclusion: By increasing laparoscopic cholecystectomies in our center we observed 0.37% incidental cases of gall bladder adenocarcinoma. Preoperative diagnosis of this cancer in early stages is very difficult and needs high degree of suspicion. The most important predictor of prognosis is the stage of the cancer at the time of surgery. To the best of our knowledge, this is the first report of incidental gall bladder cancer from Iran

Handling and pathology reporting of gastrointestinal endoscopic mucosal resection

Endoscopic mucosal resection [EMR] is a non-invasive alternative to surgery that is now frequently used for resection of early lesions in both upper and lower parts of the gastrointestinal [GI] tract. One of the main advantages of these techniques is providing tissue for histopathological examination. Pathological examination of endoscopically resected specimens of GI tract is a crucial component of these procedures and is useful for prediction of both the risk of metastasis and lymph node involvement. As the first step, it is very important for the pathologist to handle the EMR gross specimen in the correct way: it should be oriented, and then the margins should be labeled and inked accurately before fixation. In the second step, the EMR pathological report should include all the detailed information about the diagnosis, grading, depth of invasion [mucosa only or submucosal involvement], status of the margins, and the presence or absence of lymphovascular invasion. The current literature [PubMed and Google Scholar] was searched for the words "endoscopic mucosal resection" to find all relevant publications about this technique with emphasis on the pathologist responsibilities

Evaluating the frequency of MLH-1 Loss in serrated polyps of colon: a single center study from Southern Iran

Aim: Evaluation of the role of MLH-1 loss in serrated polyps of colon in a population of South of Iran

Background: There has been extensive change in classification and pathogenesis of serrated polyps of the colon during the last 10 years. The new classification is mostly based on the knowledge of the molecular pathogenesis and the rate of progression to colorectal cancer in these types of polyps. One of the most common and early lesions in molecular pathogenesis of serrated colorectal cancer is loss of MLH-1

Patients and Methods: In this study over 2 years [2012-13], 78 cases of colorectal polyps with serrated morphology resected in hospitals affiliated with Shiraz University of Medical Sciences were reclassified and investigated for MLH-1 loss by immunohistochemical method

Results: Out of the 78 colorectal polyps, 64 were classified as hyperplastic polyp [HP] and 14 as sessile serrated adenoma/polyp [SSA/P]. There was no case of traditional serrated adenoma. Three cases of SSA/P located in right colon showed dysplasia. MLH-1 loss was detected only in these 3 cases. No case of HP or SSA/P without dysplasia showed MLH-1 loss

Conclusion: SSA/P is not a common serrated polyp in our population, i.e. it is much less common than HP. Although MLH-1 loss in serrated polyps of colon is overall rare, it is fairly common in dysplastic right sided SSA/P, which confirms this molecular change as an early event in serrated carcinogenesis

Inferior vena cava reconstruction by gallbladder patch: An experimental design

Objective: To evaluate the efficacy of the gallbladder for reconstruction of the inferior vena cava in a canine model

Methods: The experimental study was conducted on 5 dogs; an oval window with a diameter of [4×1 cm] was made in the inferior vena cava and then repaired using the autologous gallbladder patch with preservation of gallbladder function. The patency and functionality of the graft were assessed macroscopically and microscopically at 2 months postoperatively

Results: All the dogs were euthanized at 2 months, showing excellent patency of the vena cava macroscopically. In the microscopic examination, all the patches were completely endothelialized. No evidence of infection and inflammation and thrombosis was noted

Conclusion: The gallbladder patch is an available and safe alternate for reconstruction of the inferior vena cava at least in animal model. However, further well designed prospective studies are needed to confirm this hypothesis

Molecular and immunohistochemical study of platelet derived growth factor receptor alpha in kit negative gastrointestinal stromal tumors; the first report from Iran

Background: Gastrointestinal stromal tumors [GISTs] are potentially malignant tumors; however their behavior and response to treatment is dependent on the type of mutation and immunohistochemical expression of antigens. It is recommended to perform routine molecular and immunohistochemical tests in KIT and platelet derived growth factor receptor alpha [PDGFRA] molecules for making decision regarding targeted therapy and prediction of the behavior of the tumor

Objectives: There has been no study from Iran regarding the PDGFRA mutational analysis in GISTs. In this study, for the first time from Iran, we performed immunohistochemical and molecular analysis of PDGFRA molecule on GISTs

Methods: In a cross-sectional study during 7 years [2008-2014] on 50 untreated non-recurrent non-metastatic newly diagnosed GISTs, molecular analysis and immunohistochemical staining for PDGFRA were performed and findings were compared with different clinicopathological characteristics

Results: During the 7 years, 50 cases of GISTs according to the above mentioned criteria were found. 17 cases were negative for KIT mutation. Of them, 15 [30%] were positive for either exon 12 or 18 mutation of PDGFRA. These cases showed more epithelioid morphology and the number of mitotic figures were lower than PDFRA negative GISTs. Also according to the criteria for risk assessment, it seems that PDGFRA mutant GISTs are rarely in the high risk category

Conclusion: PDGFRA mutant GISTs are common in Iranian population and it is recommended to perform mutation analysis for PDGFRA in every GIST with wild type KIT and epithelioid morphology

Differentiation of definitive endoderm from human induced pluripotent stem cells on hMSCs feeder in a defined medium

Background: The Definitive Endoderm [DE] differentiation using the undefined media and non-human feeders can cause contaminations in the generated cells for therapeutic applications. Therefore, generating safer and more appropriate DE cells is needed. This study compared five different methods to establish an appropriate method for inducing an efficient DE differentiation from Human Induced Pluripotent Stem Cells [hiPSCs] on an appropriate feeder in a more defined medium

Methods: Human Induced Pluripotent Stem Cells [hiPSCs] were cultured on inactivated feeders. Passaged hiPSCs, without feeder, were incubated for three days with Activin-A and different endodermal differentiation media including 1-FBS, 2-B27, 3- ITS and albumin fraction-V, 4-B27 and ITS and 5-like the third medium. The feeder cells in the first four methods were Mouse Embryonic Fibroblasts [MEFs] and in the fifth method were human adult bone marrow Mesenchymal Stem Cells [hMSCs]. DE markers FOXA2, SOX17 and CXCR4 and also pluripotency marker OCT4 were evaluated using qRT-PCR, as well as FOXA2 by the immunocytochemistry

Results: QRT-PCR analysis showed that after three days, the expression levels of DE and pluripotency markers in the differentiated hiPSCs among all five groups did not have any significant differences. Similarly, the immunocytochemistry analysis demonstrated that the differentiated hiPSCs expressed FOXA2, with no significant differences

Conclusion: Despite this similarity in the results, the third differentiation medium has more defined and cost effective components. Furthermore, hMSC, a human feeder, is safer than MEF. Therefore, the fifth method is preferable among other DE differentiation methods and can serve as a fundamental method helping the development of regenerative medicine

Early acute kidney injury based on serum creatinine or cystatin C in intensive care unit after major trauma

Background: Acute kidney injury [AKI] is a common problem in critically ill patients and is independently associated with increased morbidity and mortality. Recently, serum cystatin C has been shown to be superior to creatinine in early detection of renal function impairment. We compared estimated GFR based on serum cystatin C with estimated GFR based on serum creatinine for early detection of renal dysfunction according to the RIFLE criteria

Methods: During 9months, three hundred post trauma patients that were referred to the intensive care unit of a referral trauma hospital were recruited. Serum creatinine and serum cystatin C were measured and the estimated GFR within 24 hours of ICU admission was calculated. The primary outcome was the incidence of AKI according to the RIFLE criteria within 2[nd] to 7[th] day of admission

Results: During the first week of ICU admission, 21% of patients experienced AKI. After adjusting for major confounders, only the patients with first day's serum cystatin level higher than 0.78mg/l were at higher risk of first week AKI [OR=6.14, 95% CI: 2.5-14.7, P<0.001]. First day's serum cystatin C and injury severity score were the major risk factors for ICU mortality [OR=3.54, 95% CI: 1.7-7.4, P=0.001] and [OR=4.6, 95% CI: 1.5-14, P=0.007], respectively

Conclusion: Within 24 hours after admission in ICU due to multiple trauma, high serum cystatin C level may have prognostic value in predicting early AKI and mortality during ICU admission. However, such correlation was not seen neither with creatinine nor cystatin C based GFR

K-ras mutation in colorectal cancer, a report from southern Iran

There are very few studies about K-ras mutations in colorectal cancer [CRC] from developing countries such as Iran. It is therefore essential to conduct studies to learn about the molecular signature of such tumors, allowing the determination of an appropriate management plan. In the present study, we aimed to determine the frequency and types of K-ras mutations among patients with CRC in Iran. Formalin-fixed paraffin-embedde

P63 and Ki-67 expression in dentigerous cyst and ameloblastomas

Statement of the Problem: P63 gene is a member of TP53 and its homologous gene family. Its expression was observed in some odontogenic lesions, more expression in aggressive lesions

Purpose: This study aimed to investigate the possible diagnostic impact of P63 protein on dentigerous cysts and various types of ameloblastoma. Its expression with Ki-67 proliferation marker was also compared

Materials and Method: This cross-sectional retrospective study was enrolled on 25 cases of dentigerous cyst including 21 unicystic ameloblastomas and 17 conventional ameloblastomas. The expression of P63 and Ki-67 was assessed by immunohistochemical [IHC] examinations. Data were analyzed by employing Mann- Whitney and correlation coefficient tests

Results: P63 expression was significantly higher in ameloblastoma than unicystic ameloblastoma and dentigerous cysts. There was no significant difference between unicystic ameloblastoma and dentigerous cyst in P63 expression. A 90% cut-off point was obtained for basal layer which gave 88% sensitivity and 78% specificity to distinguish more invasive lesions from others. There was not any correlation between P63 and Ki-67 immunostaining in the three study groups

Conclusion: More aggressiveness and more invasiveness of odontogenic lesions depicted higher rate and also more intensive expression of P63. Moreover, the expression of P63 protein had not any correlation with Ki-67 protein in dentigerous cysts and ameloblastomas

Antibacterial and toxic effect of hydrogen peroxide combined with different concentrations of chlorhexidine in comparison with sodium hypochlorite

Statement of the Problem: Hydrogen peroxide [H[2]O[2]] has been suggested to be used in sequence or in combination with chlorhexidine [CHX] to enhance the antibacterial activity against Enterococcus faecalis, but there is no research in the literature on the safety and effectiveness of this irrigation protocol

Purpose: This study aimed to assess the cytocompatibility and antibacterial activity of different concentrations of CHX combined with H[2]O[2] in comparison with the activity of 5.25 and 2.5% sodium hypochlorite [NaOCl]

Materials and Method: Different concentrations of H[2]O[2] [10, 5, 3 and 1%] were exposed to the PDL cells. Then, the solution with minimal cytotoxicity was selected [3% H[2]O[2]]. The cytocompatibility and antibacterial activity of 0.1, 0.2, 1 and 2% CHX combined with 3% H[2]O[2] were evaluated and compared with 5.25 and 2.5% NaOCl. The differences in the mean viability of PDL cells were evaluated by one-way ANOVA. Kruskal-Wallis and post-hoc Dunn's tests were adopted to compare the antibacterial activity of the solutions against E.faecalis

Results: The viability of PDL cells was lower when treated with 5.25 or 2.5% NaOCl than all combinations of CHX and H[2]O[2]. There was no significant difference in the antibacterial activity of the solutions against E.faecalis, except for the 0.1% CHX + 3% H[2]O[2] combination, which had significantly lower efficacy than other groups

Conclusion: All combinations of CHX and H[2]O[2] [used in this study] except 0.1% CHX + 3% H[2]O[2] were efficient irrigants against planktonic E.faecalis and had a better cytocompatibility with PDL cells than 5.25 and 2.5% NaOCl

Topical simvastatin enhances tissue regeneration in full-thickness skin wounds in rat models

Wounds and wound healing have always been one of the most important subjects that experimental researches were dedicated to. Simvastatin has been used for long as a common lipid lowering agent which was reported to have some pleiotropic effects such as antioxidation, anti-inflammation and immunomodulation. In this study we aimed to determine the effect of simvastatin on wound healing process in laboratory rats by means of stereological and histopathological analyses. 36 male Sprague-Dawley rats [220 +/- 20 g] with a 1 cm[2] circular full-thickness wound on their back were divided into three groups: SS group that received a gel with 2% concentration of simvastatin; UW group that received no treatment but daily irrigation with normal saline; Base group that was treated with the gel base. Duration of the study was 12 days and at the end, wound closure rate, grade of inflammation, granulation-tissue formation, ulceration, epithelization, fibroblast proliferation, collagen-bundles synthesis, and vascularization were determined. Outcome of this study revealed that Simvastatin improves the wound healing by its anti-inflammatory and epithelization induction effect as well as statistically significant induction of fibroblast proliferation and collagen bundle synthesis which were reported by our stereological and histopathological investigations. Results of the present study demonstrated that topical Simvastatin enhances the wound healing process through affecting different aspects of tissue regeneration; however, further researches are needed to find the exact mechanism, advantages and disadvantages of this chemical agent

57 year old woman with polypoid gastric mass

A 57-year-old woman was referred with chief complaint of pallor and lethargy for 9 months. Laboratory investigation was notable for anemia and occult blood in the stool exam. Her past medical history was unremarkable. Endoscopy of the upper gastrointestinal tract showed a large fungative reddish mass measuring 7 x 6 cm in the body of the stomach and multiple variable size polyps in the antrum [figure 1]. The patient underwent hemi-gasterectomy. Billroth I. Gross examination of the stomach showed a large polypoid mass measuring 8x7x6 cm [figure 2] with erosive surface in some areas

Interposition of ileal J-pouch for rectum reconstruction in dog

Background: The gold standard of the management of rectal cancer in the middle and lower parts is low anterior resection with coloanal anastomosis. About 50% of the patients undergoing this procedure might experience some complications because of the low capacity of the neorectum. The aim of this study was to evaluate ileal J-pouch interposition as a neorectum between the anal canal and the remaining colon in comparison to coloanal anastomosis and transverse coloplasty

Methods: Twelve dogs, weighing 23-27 kg, were divided into three groups. After laparotomy, the volume of the primary rectum was measured so that it could be compared with that of the neorectum at the end of the study. After rectal resection in Group A, the colon was directly anastomosed to the anus. In Group B, a 5-cm longitudinal incision was made 2 cm proximal to the anastomosis and was sutured transversely [coloplasty]. In Group C, a 5-cm ileal J-pouch was interposed between the colon and anus. After 8 weeks, the neorectum was evaluated for volume, radiology, and pathology

Results: All the samples were alive until the end of the study. The healing of the anastomotic lines was acceptable [pathologically] in all. The mean volume expansion was 20.9% in Group A, 21.7% in Group B, and 118.2% in Group C, with the latter being significantly higher than that of the other groups [P=0.03]. Colon J-pouch and coloplasty after proctectomy in some situations have not been performable. This study evaluated the performance of ileal J-pouch interposition

Conclusion: This study showed that ileal J-pouch interposition might produce an acceptable reservoir function and that it seems feasible and safe in selected cases

Prolonged hemoptysis caused by primary pulmonary epithelioid hemangioendothelioma; a case report and review of the literature

Epithelioid hemangioendothelioma is a vascular tumor with an intermediate malignant potential. This tumor is very rare in the lung parenchyma, and most of the previously reported cases have been asymptomatic. There is no standard therapy for this tumor and prognosis in the previous reports has been variable. Herein we report our experience with a 60-year-old woman presenting with hemoptysis and multiple lung consolidation, leading to a diagnosis of epithelioid hemangioendothelioma after surgical resection and pathological examination. After surgery and chemotherapy, the patient had an acceptable course