Iranian Journal of Pathology

Abstract Background & Objective: Reproductive toxicity of cadmium (Cd) as an environmental toxicant has been proved in animals and humans. Exposure to Cd impairs testes organs and can reduce male fertility. The present study was designed to investigate the spectrum of histopathological changes in testicular tissue focusing on Sertoli cells in rats following Cd intoxication.
Methods: In the present experiment, acute testicular toxicity was induced by an intraperitoneal injection of 1.2 mg/kg CdCl₂ to the animals in the test group, while the control group received normal saline. After 52 days, the animals were euthanized, and testicular tissue was stained by Hematoxylin and Eosin. In addition, immunohistochemical staining was performed on Sertoli cells for Wilms' Tumor, Melan-A, and CD99 to evaluate histopathological changes.
Results: Cd caused significant alterations in seminiferous tubules with varying effects on the patterns of spermatozoa production. These histopathological changes were significantly higher in the Cd group, compared to the control group.
Conclusion: The Cd-induced stepwise spectrum changes included sloughing, disorganization, hypospermatogenesis, spermatic cell arrest, germ cell hypoplasia, Sertoli cell-only pattern, fibro-hyalinized seminiferous tubules, and calcification. Sertoli cells accumulated and created multinucleated giant cells in the seminiferous tubules during the atrophic process, which could be dependent upon Sertoli cells viability and function. 

Abstract Background & Objective: Epithelial-Mesenchymal transition (EMT) is known to be a possible mechanism in tumor progression; however, there is insufficient evidence to support the contribution of this process in human cancers. The present study aimed to evaluate the expression of EMT markers in normal oral epithelium and oral squamous cell carcinoma and also correlates with some clinicopathological parameters.
Methods: This study was conducted on 70 samples, including 20 cases of normal epithelium and 50 cases of Oral Squamous cell Carcinoma (OSCC). To examine the expression level of these proteins, immunohistochemical staining was performed for samples using E-cadherin and N-cadherin monoclonal antibodies.
Results: Reduced expression of E-cadherin was observed in 74% of OSCC and 15% of normal epithelium samples; this difference was statistically significant (P˂0.000). With the progression of SCC from well towards poor differentiation, the E-cadherin expression decreased; however, this difference was not statistically significant (P=0.642). Normal epithelial specimens were negative for N-cadherin expression in 75% of cases, whereas OSCC specimens showed high expression of N-cadherin in 46% of cases, this difference was statistically significant (P=0.01). Although 62.5% of poorly differentiated OSCC showed high expression of N-cadherin, the difference between the histopathological grades was not significant (P=0.586). No significant relationship was found between markers expression and patient’s age, gender, and tumor location.
Conclusion: This study showed that OSCC tissues showed high EMT phenotype (reduced E-cadherin expression and high expression of N-cadherin) compared to normal oral mucosa which may indicate the possible key role of EMT mechanism during oral carcinogenesis. 

Abstract Background & Objective: Vascular Endothelial Growth Factor (VEGF) is one of the newer molecular markers that acts as a central mediator of tumor angiogenesis and is essential for tumor growth, progression, and metastasis. So anti-angiogenic drugs can be used as anticancer therapy. Treatments with anti-VEGF (Bevacizumab) therapy have been proved to improve relapse-free survival in many tumors. Urinary bladder tumor has become emerging cancer globally among elderly individuals. So, the identification and development of novel biomarkers for effective treatment of urinary bladder carcinoma is essential. The present study aimed to investigate the immunohistochemical expression of VEGF in urothelial carcinoma of urinary bladder and to assess its association with tumor grade and muscle invasiveness.
Methods: This cross-sectional study was conducted in the Department of Pathology, Chittagong Medical College, Chattogram from September 2018 to August 2020. Fifty-six formalin-fixed paraffin-embedded tissue blocks of urinary bladder carcinoma were prepared for both histopathological and immunohistochemical examination. Each slide was evaluated by at least two pathologists.
Results: Weak to strong positive expression of VEGF were observed in 52 cases (92.86%). The proportion of tumors positive for VEGF expression was higher among patients with high grade and non-muscle invasive bladder carcinoma. 
Conclusion: We found that VEGF expression has a significant association with tumor grade and an inverse association with muscle invasion. These findings may be useful for selecting the subset of patients likely to respond to anti-VEGF targeted therapy. 

Abstract Background & Objective: Acral melanoma (AM) is a common type of cutaneous melanoma that occurs in the skin of the palms, soles, and nail beds. This malignancy, like other types of cancer, has different genetic alterations. To date, despite decades of research the roles of oncogenic BRAF mutations in the pathogenesis of AM has not been fully clarified. The present study was designed to identify V600E mutation in patients with AM from the south of Iran. 
Methods: The samples were collected from the pathology lab archive of Shiraz University of Medical Sciences (2015-2020). A total of 41 patients with primary invasive AM underwent excisional biopsy or amputation were collected to evaluate BRAF V600E mutation using Polymerase Chain Reaction (PCR) and Sanger sequencing.
Results: Total number of 41cases (21 male and 20 female) and age range of 34-87 years were enrolled. The histological subtypes were 24 acral lentiginous melanomas (ALM), 10 cases of nodular melanoma (NM), and 7 cases of superficial spreading melanoma (SSM). In our study, only one case (a 44-year-old male with nail bed AM and the histological subtype of acral lentigenous melanoma) showed BRAF-V600E mutation. 
Conclusion: These findings suggest that the population of our interest showed a very low prevalence of this mutation providing novel insights into the pathobiology of AM and its related treatment. 

Abstract Background & Objective: The role of Epstein-Barr Virus in development of breast cancer is frequently studied. In this regard, miRNAs are among the contributing elements in the molecular pathophysiology of EBV-related diseases. In addition, a growing number of host miRNAs are believed to be implicated in pathogenesis of breast cancer. MiR-218 is a tumor suppressive miRNA that is subjected to dysregulation in various EBV-associated cancers. We aimed to investigate the frequency of EBV and its relationship with expression status of tumor suppressive miR-218 in breast cancer and adjacent normal tissue.
Methods: A total number of 51 fresh malignant breast cancer tissues (cases) and their adjacent normal tissues (controls) were collected. Nested-PCR and RT-qPCR were set to identify EBV frequency and miR-218 expression in cases and controls, respectively.
Results: Out of all samples, 6.8% (7/102) comprising 11.6% (6/51) in malignant tissues and 1.9% (1/51) in normal control tissues were positive for EBV (P<0.05). Quantitative data showed that miR-218 was significantly downregulated in malignant tissues compared to control tissues (P<0.0001). In addition, reduced expression of miR-218 was associated with adverse clinical outcomes, metastasis, and higher grades of malignancy. Given the presence of EBV, lower expression of miR-218 was observed in breast cancer group in comparison with normal group (P<0.05). 
Conclusion: Our results raise the possibility of the relation between EBV infection and miR-218 downregulation in breast cancer and propose further investigations in this regard. 

Abstract Background & Objective: Escherichia coli (E. coli) is a leading cause of urinary tract infections becoming resistant against beta-lactams and cephalosporins through different mechanisms, including ESBL production due to the presence of ESBL specific genes, including blaCTX-M and blaTEM. The purpose of the present study was to detect the uropathogenic E. coli strains producing the ESBL.
Methods: A total of 100 isolates of uropathogenic E. coli were randomly selected in a period of 6 months and their resistances to a number of antibiotics including amoxicillin, amikacin, gentamicin, ciprofloxacin, ceftazidime, cefotaxime, ceftriaxone, ceftizoxime, nalidixic acid, and nitrofurantoin were determined. Then, DDT test was used to detect the presence of ESBL. Finally, the presence of blaCTX-M and blaTEM resistance genes was analyzed by PCR method.
Results: The resistance profile of bacterial isolates to the antibiotics was as follows: amoxicillin: 16.7%, amikacin: 7.8%, gentamicin: 20.3%, ciprofloxacin: 35.5/%, ceftazidime: 35.0%, cefotaxime: 40.0%, ceftriaxone: 41.3%, nalidixic acid: 64.0%, nitrofurantoin: 9.7%, and ceftizoxime: 100%. Of these, 28 isolates (28%) were reported to be resistant to cefotaxime, ceftazidime, and ceftriaxone. In DDT test, 21 ESBL positive cases (21%) were detected. PCR results showed that the presence of blaCTX-M and blaTEM genes in the isolates were 21% and 20%, respectively. 
Conclusion: Regarding the production of ESBL by some E. coli isolates, phenotypic detection of ESBL-producing isolates is routinely suggested in the laboratories. Likewise, the treatment regimen should be selected regarding the ESBL production to avoid treatment failure. 

Abstract Background & Objective: Liver lesions are difficult to diagnose and to differentiate primary from metastatic carcinoma, while Biopsy has its limitations. Cell block technology is easily accessible with high diagnostic accuracy. Our aim is 1) To find the role of cell block technology as an alternative to biopsy in identifying liver lesions; 2) To find the efficacy of cell block along with immunohistochemistry (IHC) and ancillary studies in differentiating primary from metastatic lesions; 3) To identify the site of origin of metastatic lesions. This is a descriptive study undertaken in two tertiary care hospitals over a period of three years.
Methods: Retrospective review of adequate samples from fine needle aspirations from liver lesions under radiological coverage, converted into cell block was done. IHC was applied as needed. Usefulness of cell block preparation was evaluated, and the final diagnosis correlated with the biopsy results.
Results: Analysis of 323 cases found sensitivity of 98.75% and positive predictive value of 99% for all lesions. Sensitivity for metastatic carcinomas was slightly more than hepatocellular carcinoma. However, accuracy of cell block results for individual metastatic lesions and site of origin was less. IHC and morphological pattern worked as an important adjunct in the final diagnosis. On the other hand, contribution of viral markers as a supplement in the final work up was ambiguous. 
Conclusion: High precision of validity results of cell block technology in comparison with biopsy highlights its pivotal role in conjunction with supportive tests for diagnosing and differentiating liver lesions as well as identifying primary sites in liver metastasis. 

Abstract Background & Objective: The ability of Pseudomonas aeruginosa to form biofilm has an important role in establishment of chronic phase of infections. Biofilm formation can be affected by antibiotics sub-MIC concentrations. The principal aim of the present study was to evaluate the effect of gentamicin at sub-MIC concentrations on biofilm formation in 100 Pseudomonas aeruginosa clinical isolates.
Methods: Determination of minimal inhibitory concentration of gentamicin for clinical isolates was done using micro broth dilution method. The amount of biofilm formation in the treated and untreated isolates with gentamicin sub-MIC (1/2&1/4MIC) concentrations was evaluated using microtitre plate assay. pelA and pslA genes were detected in clinical isolates by PCR method.
Results: 99% of clinical isolates were biofilm producer. Different changes in amount
of biofilm formation were observed in the treated clinical isolates with sub-MIC concentrations of gentamicin. Two dominant changes were observed in 80% of clinical isolates. These concentrations had inhibitory effect on biofilm formation in 46.4% of isolates and caused a significant decrease in its amount. While in 31.3% of the isolates, the biofilm formation was significantly increased. The frequency of pelA and pslA genes among clinical isolates was 100%. 
Conclusion: gentamicin sub-MIC concentrations cause different changes on biofilm formation of Pseudomonas aeruginosa clinical isolates. Therefore, further studies are needed for discovering new treatment strategies and using sub-MIC concentrations of the antibiotic in prevention and treatment of Pseudomonas aeruginosa infections. 

Abstract Background & Objective: Some certain markers, including prostatic specific antigen (PSA), are being used to screen prostate cancer (PC), but none of them have sufficient sensitivity and specificity for evaluation of prognosis. Currently, genetic variants have found their place in the prognosis of PC. ETS-related gene (ERG) expression and its intensity have contradictory evidence regarding ERG expression with PC incidence or associating outcome. Our purpose was to survey the relationship of ERG expression and its intensity with PC and relative clinical outcome.
Methods: We studied the immunohistochemichal (IHC) expression of ERG in 101 radical prostatectomy specimens with PC of different histologic grades. All samples were chosen from pathology department of Sina hospital in Tehran-Iran from 2011 to 2018.  Positive ERG expression and its association with Gleason score, preoperative PSA, metastasis status, stage and grade of tumors was evaluated.
Results: In total, ERG expression was observed in 42 cases (41.58%) and of these, 7 (16.66%) were categorized as weak, 13 (30.95%) moderate and 22(52.38%) as strong. There was no significant correlation between ERG expression and age, preoperative PSA, Gleason score, lymph node involvement, metastatic pattern, stage, and grade of the tumor (P>0.05). ERG expression frequency in the two groups of survived and expired patients was 42.85% and 0%, respectively; despite the noticeable difference, it was not statistically significance (P=0.264).
Conclusion: Evaluation of ERG expression and its intensity may have no essential role as an acceptable prognostic factor in Iranian’s population for anticipating whether PC itself or the outcomes accompanied. This relation is vigorously under the influence of geographical/ethnical features. 

Abstract Background & Objective: One of the major genetic causes of recurrent spontaneous abortions is parental chromosomal abnormalities. The objectives of the study were to determine, compare and analyze the incidence and distribution of chromosomal abnormalities in couples with recurrent miscarriages from Northeastern Iran.
Methods: This study was conducted at Ghaem Hospital, Mashhad, Iran. We evaluated karyotype results of 608 couples with history of recurrent spontaneous abortion. The standard method was used for culturing peripheral venous blood lymphocytes.
Results: Chromosome aberrations were detected in 43 patients (3.54%), including 25 females and 18 males. Structural chromosomal abnormality was detected in 40 cases, including balanced translocations (25 cases), robertsonian translocations (4 cases), inversions (10 cases) and numerical chromosome aberrations (3 cases). Polymorphic variants were observed in 22 individuals.
Conclusion: The frequency of chromosomal abnormalities in couples with Recurrent Spontaneous Abortion (RSA) in our study is 3.54%. Reciprocal translocation, pericentric inversions, robertsonian translocations, and numerical abnormality observed among couples who had experienced recurrent spontaneous abortions and that these couples might benefit from cytogenetic analysis. 

Abstract Background & Objective: Pseudomonas aeruginosa is an opportunistic pathogen and one of the most common causes of nosocomial infections. This bacterium's antibiotic resistance to the common fluoroquinolone antibiotics, especially ciprofloxacin, is due to mutations in the gyrA and parC genes. This study aimed to investigate the effect of the mutation in (gyrA, parC) on ciprofloxacin resistance in clinical isolates of Pseudomonas aeruginosa.
Methods: A total of 140 clinical samples were collected from hospitals. The samples were identified by standard biochemical tests, and the antibiotic resistance was investigated by the disk diffusion method. DNA was extracted from 30 isolates, and PCR was performed. PCR-sequencing was carried out to assess gyrA and parC mutations in drug-resistant isolates. NCBI-Blast and MEGA7 software was used to analyze the nucleotide sequences.
Results: 30 clinical isolates were 80% resistant to ciprofloxacin; meanwhile, in 21 samples, mutations were observed. 87/5% of mutations were related to gyrA (Thr83 → Ile), 79/16 % parC (Ser87 → Leu), and 4/18% (Glu91 → Lys). The antibiotic resistance to ciprofloxacin and mutations in gyrA and parC genes in resistant isolates are significantly related to each other (P<0.05). 
Conclusion: The mutations in the gyrA and parC genes play an essential role in resistance to ciprofloxacin in clinical isolates of Pseudomonas aeruginosa. 

Abstract Background & Objective: Differential diagnosis between cholangiocarcinoma (CCA) and metastatic pancreatic ductal adenocarcinoma (PDA) in the liver is difficult and so far, no specific immunohistochemical marker is reported to differentiate these two tumors. Considering the existing literature, the level of expression of Annexins (Annexin A1, 10 and 13) have been studied for differential diagnosis between these two tumors by molecular methods and promising results have been reported. Therefore, in this study, we tried to investigate the immunohistochemical value of these three Annexins for the differential diagnosis of CCA and PDA in the liver.
Methods: The articles that reported the research subject in 10 years (2009-2019), including 45 cases of CCA and 50 cases of metastatic PDA in the liver were evaluated considering the presence or absence of AnnexinA1 (ANXA1), Annexin A10 (ANXA10) and Annexin A13 (ANXA13) expression by immunohistochemistry, were investigated.
Results & Conclusion: This study showed, ANXA1 was positive both in PDA and CCA, ANXA10 was positive in ~60% of PDA cases and ~40% of CCA cases, and ANXA13 was mostly negative in both groups. The best sensitivity was found in cytoplasmic and nuclear ANXA1 (80% and 84%, respectively) to distinguish PDA from CCA and vice versa. The best specificity was observed in ANXA10 and ANXA13 to distinguish PDA from CCA. Also, ANXA13 had the best specificity to distinguish CCA from PDA. Our investigations showed that, ANXA1 probably can classify positive cases correctly, but it cannot discriminate PDA from CCA. ANXA10 had fair sensitivity and specificity to discriminate PDA from CCA. ANXA13 apparently had a high specificity that can help to narrow-down the differential diagnoses. 

Abstract Background & Objective: Pathology education conventional methods have been disrupted by the Corona-Virus Disease 2019 (COVID-19) outbreak, compelling a re-evaluation of the means of educational interactions from the undergraduate to the postgraduate level. This commentary explores how the COVID-19 outbreak has challenged pathology education.
Methods: We reviewed the current challenges and determined the potential implications of virtual technologies on modern pathology education for the future of pathology competency learning and assessment.
Results & Conclusion: The challenges are partly due to transferring from in-person teaching to a virtual education. Other reasons are shifting away from discipline-based teaching to organ-system based in medical curriculum and additional pressures on pathology faculties, such as increased demand for pathology services, lack of time, and learning resources. Keeping the national standards in pathology education even in the constant disruptions from pandemic outbreaks are current challenges. Pathology expertise will need to use emergent technologies in providing educational material to ensure quality pathology education. However, virtualization of pathology education produces a value of digital pathology and web-based pathology training materials. Medical students could review clinical cases remotely with their supervisors and gain the pathology competencies necessary for clinical practice.
We need new innovative strategies, and we suggested the following steps to take advantage of the current opportunity to meet the challenges: evaluating the available digital training materials for formal pathology education, investing in the virtual infrastructure for competency-based pathology education, expanding student/residents exposure to pathology educational cases through virtual platforms; applying digital pathology solutions for virtual pathology education. 

Abstract Background & Objective: Diabetes is a metabolic disease and is associated with failure of various organs. Macrophage migration factor (MIF) and matrix metalloproteinase (MMP-9) are two of the most important factors in the pathogenesis of diabetes.
Methods: In this descriptive-analytical study, 30 patients with type 2 diabetes mellitus from Hamadan Diabetes Center were selected by convenience sampling. Moreover, 30 healthy first-degree relatives and 30 unrelated non-diabetics, were examined for MMF and MMP-9 and their variations based on age, gender, body mass index (BMI) and hemoglobin A1C.
Results & Conclusion: The mean and standard deviation of MIF in diabetic patients, and relatives and non-relatives of diabetic patients were 592.87±78.19, 131.82±88.27 and 94.63±23.88, respectively (P<0.001). The mean and standard deviation of the MMP-9 in diabetic patients, and relatives and non-relatives of diabetic patients were 2570.64±2220.03, 918.57±650.08 and 629.09±288.32, respectively (P<0.001). MIF and MMP-9 did not have a significant relationship with age, sex, duration of disease and BMI. However, we observed a direct and significant correlation between hemoglobin A1C and the level of MIF and MMP-9 (P<0.001). In patients with type 2 diabetes, serum levels of MMP-9 and MIF, consistent with HbA1c, increase with no significant association with age, sex, BMI and duration of diabetes. 

Abstract The incidence of pericardial epithelioid angiosarcoma is rare. Angiosarcoma of pericardium may coat the pericardium in a diffuse fashion. Diagnosis of an angiosarcoma is challenging and may be easily mistaken as constrictive pericarditis. Herein, a case of primary pericardial angiosarcoma is reported in a 16-year-old female. Patient presented with chest pain and dyspnea on exertion, regarded as constrictive pericarditis. Pericardectomy was performed and histopathologic examination showed pleomorphic epithelioid cells exhibiting hyperchromatic nuclei, prominent nucleoli and eosinophilic cytoplasm arranged in sheets and occasionally lined irregular vascular spaces. Moreover, immunohistochemical staining revealed that tumor cells were positive for CD31 and vimentin. The patient received chemotherapy with adriamycin, ifosfamide, and mesna. Unfortunately, the patient died of cardiac involvement and pleural metastases less than three months following the operation. Primary pericardial angiosarcoma is rare and difficult to diagnose, especially epithelioid variant. Immunohistochemical assessment is required to confirm the final diagnosis.

Abstract Myofibroblastoma (MFB) of the breast is an uncommon entity of benign spindle neoplasms of the breast. This tumour possesses a broad spectrum of histomorphological patterns. Distinguishing of myofibroblastoma variants from malignant mimics of this benign neoplasm is essential for pathologists to avoid further invasive surgical procedures. In this article, we report the clinical, morphological, and immunohistochemical features of three cases, including two females and one male patient with mammary myofibroblastoma with emphasis on the histomorphological findings. As there is not yet enough information about MFB, more reports of MFB are still required to more clarify the pathogenesis and potential predisposing factors of this rare type of breast tumours.

Abstract Fibroepithelial polyps of the vagina (FEPV) are rare entities which normally manifest as one or more painless polyps sometimes with symptoms such as bleeding, vaginal discharge, and discomfort regarding the size of the mass. Despite their benign nature, they can be confused with other vaginal tumors due to their abnormal histology. In this report, we present a case of a 44-year-old woman with a giant pedunculated and symptomatic polyp of the vagina with anterior vaginal wall prolapse. The treatment method included a simple local excision of the polyp and anterior vaginal compartment repair. Histopathological examination revealed a polypoid lesion covered by squamous epithelium containing a central fibrovascular core without atypia. The patient experienced an uneventful postoperative recovery, with no complication, which implies that surgery is the most effective modality for managing such tumors.

Abstract Primary leiomyosarcoma in testis is an uncommon tumor with few cases reported. It generally develop  after radiotherapy or long term taking anabolic steroid medication . We report a  53-year-old male patient with primary testis leiomyosarcoma  who presented with painless testicular  enlargement without any known  predisposing factors. Ultrasound revealed a large heterogeneous left testicular solid lesion. Alpha-fetoprotein (AFP) and beta-human chorionic gonadotrophin (beta-HCG) levels in serum were normal. Left radical orchidectomy following with histology assessment established a diagnosis of primary leiomyosarcoma of testis. No data of cancer metastasis was established. The patient didn’t receive any adjuvant therapy. There wasn’t any evidence of recurrence after 1 year follow-up. Leiomyosarcoma must be one of the differential diagnoses of seronegative tumors in testis. The motivation for this paper is the extreme infrequency of the situation and the differential diagnosis by all expansive inguinoscrotal tumors.

Abstract Chemotherapy of ovarian tumors has some effects on morphology of stromal cells also residual tumoral cells which can cause problems in diagnosis of tumor type and staging of it .In some cases ,it can makes problem in diagnosis of residual epithelial tumor from other differential diagnosis such as spindle cell tumors like sarcoma.We had a case which had previouse chemotherapy. Her ovarian tumor had plump spindle cells with pleomorphism similar to spindle cell tumors.Finally ,in these cases ,Immuno histochemisteric study can differentiate correct diagnosis from differential diagnosis.