Iranian Journal of Pathology

Abstract The ileum undergoes endoscopic biopsy much more than before. Most of these biopsies are either completely normal or show non-specific changes. Nevertheless, in some diseases, ileal biopsy is diagnostic, and in some cases it may be the only anatomical location of the disease. Endoscopically normal mucosal biopsy is unlikely to provide useful diagnostic information and is not routinely recommended. However, in the presence of ileitis, ulcers, or erosions, biopsies can be very helpful. Ileitis might be induced by various conditions including infectious diseases, vasculitis, medication-induced, ischemia, eosinophilic enteritis, tumors etc. The conclusive cause of the condition is proposed by a comprehensive clinical background and physical examination, laboratory investigations, ileocolonoscopy, and imaging findings. Ileoscopy and biopsy are mainly useful in correctly selected cases such as patients that present with inflammatory diarrhea and endoscopic lesions. This review is meant to provide a simple algorithmic approach to the ileal biopsy samples. There are several boxes that give diagnostic clues and an idea behind the categories of ileal disorders.
This review is based on the reviewed literature and the authors' experiences. We have summarized different histological patterns in the ileal biopsy specimens that can be used in the diagnosis of inflammatory disorders of the ileum.
This review provides an algorithmic approach to the clinicopathological features of inflammatory disorders of the ileum with a brief discussion of some important related issues.

Abstract The etiology of parathyroid carcinoma (PC) is largely unknown. Associations have been made with several inherited syndromes and with specific genetic lesions. The management of PC is challenging for clinicians. The complexity of molecular phenotypes increases with tumor aggressiveness. Lack of parafibromin on immunohistochemistry staining and HRPT2 mutation present capable consequences in differentiating carcinoma from adenoma. Lack of parafibromin expression, the gene product of HRPT2 is now used as a diagnostic, prognostic and predictive marker for parathyroid carcinoma. The epigenetic alteration, for example, DNA methylation and modifications in the chromatin structure, are known as significant events that are the reason for parathyroid tumorigenesis. We suggest that adjuvant genetic and epigenetic target therapy should be considered in treating PC patients. 

Abstract Background & Objective: It is noteworthy that vast data links NETs to human arterial thrombosis. In the current study, extracellular neutrophil networks and macrophage polarization were assessed in the area outside and inside the Carotid artery stenosis.
Methods: The sample of Carotid plaque of the patient was divided into two halves with a transverse incision; the terms inner part and outer part were used for the plaque's inner part and the adjacent area. Samples were sorted in 10% formalin for CD163, CD11c, MPO, and histone H3 immunohistochemistry assessment, while part of the sample was stored at -80°C for western blotting assay with PDA4 marker.
Results: Results of this study showed that the extracellular neutrophil in the inner part of the Carotid plaque was significantly increased (P<0.0001), while the number of M1 and M2 macrophages was higher in the inner part compared with the outer part of the Carotid plaque (P<0.0001).

Abstract Background & Objective: Female breast cancer is one of the most prevalent malignancies among women. The critical step in managing breast cancer is to diagnose it accurately. Hence, peripheral blood-based tests are one of the most favorable and less invasive methods to study. Recent studies investigated and evaluated the inflammation parameters such as neutrophil: lymphocyte ratio (NLR), the platelet: lymphocyte ratio (PLR), and the C-reactive protein (CRP) levels. The elevation in mentioned parameters was proposed as a key factor in cancer progression. The main goal of this study was to investigate the association of NLR, PLR, and CRP levels in patients with breast lesions.
Methods: The NLR, PLR, and CRP levels were calculated from 200 female patients with either benign or malignant lesions.
Results: The cut-off values of NLR, PLR, and CRP were 1.24, 96, and 10.36 mg/L, respectively. A significant difference in NLR (P<0.001), PLR (P<0.001), and CRP levels (P<0.001) were observed between the two major studied cohorts.
Conclusion: Elevated NLR, PLR, and CRP levels could predict the presence of malignancy. In addition to the low cost and properties of the mentioned methods, utilization of this data could facilitate and improve clinical decision-making for treatment. 

Abstract Background & Objective: Acute myeloid leukemia (AML) is a hematopoietic malignancy caused by genetic abnormalities. These days, molecular and genetic factors are usually used as diagnostic and prognostic markers. FLT-3 is one of the most known diagnostic factors in AML. MDR1 gene belongs to the ATP binding cassette family; it is known as one of the chemotherapy-resistant causes of AML. We aimed to study FLT-3ITD mutations and their association with MDR1 gene expression in AML individuals.
Methods: For investigation, 80 AML individuals and 20 healthy controls were selected. This study was done in the cancer molecular pathology research center of Mashhad University of Medical Sciences (MUMS), Iran during 2017-2019. FLT3-ITD mutation was assessed by polymerase chain reaction (PCR); Real-time quantitative PCR was performed to measure the amount of MDR1 gene expression. Bone marrow and blood smears of patients were evaluated in terms of morphology. SPSS 16.0 was used for data analysis.
Results: FLT3-ITD mutation and MDR1 overexpression were found in 18.8% and 23.8% of AML patients, respectively. Statistical analysis did not show any relations or association between these two markers. Cuplike morphology was observed in blast cells in 21.25% of AML cases, which was associated with FLT3-ITD mutation presence.
Conclusion: FLT-3 and MDR1 do not affect each other. It is suggested to perform survival studies to determine the exact role of MDR1 overexpression in drug resistance issues.

Abstract Background & Objective: The TSH reference range's validity affects the thyroid dysfunction diagnosis. The primary objective of this study is to determine the reference range, which is established according to age and region.
Methods: The data were collected retrospectively from people over the age of one who visited Motahari Clinic for routine health checkups between August 2017 and October 2019. TSH, T4, T3, personal drug usage, and thyroid history were collected. After excluding subjects with thyroid diseases and outliers, a list of 1392 participants was analyzed. Hormone intervals of men and women ≥1 year old have been determined using the non-parametric method.
Results: The non-disease subjects' TSH, T3, and T4 reference ranges were 0.64 to 5.94 lU/mL, 0.91 to 2.47 ng/dL, and 5.53 to 12.48 g/dL, respectively. According to this range, total thyroid dysfunction prevalence in our study in children was 8.94%. There was no significant difference between TSH, T4 level, and sex in the non-disease population (P=0.46 and 0.13, respectively), but there was a statistical difference between sex and T3 (P =0.03). Our study also illustrates that for subjects under 18 years old and above it, hormones (TSH, T3, T4) concentration is statistically different (P≤0.001).
Conclusion: We found a statistical difference between hormone values after and before age 18 (P=≤0.01); therefore, it is not appropriate to use the same reference range for children under age 18 and adults. There was male dominance in the population 1-18 years old. 

Abstract Background & Objective: Breast cancer is the most common cancer in developed and developing countries. This study mainly addresses the issue of an equivocal result in IHC, which then needs further assessment if the patient has to receive targeted therapy. The study aimed to detect the expression of Her2/neu protein in breast cancer by immunohistochemistry (IHC) and Fluorescence in situ Hybridization (FISH) and evaluate concordance and discordance between the two methods. Also, the clinicopathological parameters in these patients were studied in association with ER, PR, HER-2, and Ki-67.
Methods: This study was conducted on 34 female carcinoma breast specimens, including core biopsies and mastectomies. Each case underwent histopathological and immunohistochemical studies for (Estrogen Receptor) ER, (Progesterone Receptor) PR, (Human Epidermal growth factor Receptor 2) HER-2, and Ki-67. In addition, FISH was done on all the samples to detect Her2 gene amplification.
Results: The overall concordance between the two tests was 79.41% while the concordance between the two tests in equivocal cases, was 14.3%. ER/PR expression and HER-2 amplification were inversely associated. Also, Ki-67 expression was not associated with the side size of the lesion, lymphovascular invasion, and lymph node metastasis. Age less than 50 at presentation and infiltrating ductal carcinoma histological type showed increased proliferation index.
Conclusion: The highest concordance between FISH and IHC was noted in IHC positive and negative cases, whereas IHC equivocal cases showed low concordance. FISH accurately determines the assessment of HER2 expressions in equivocal cases. 

Abstract Background & Objective: Papillary thyroid cancer (PTC) is the most common primary cancer originating from thyroid follicular cells. The aim of this study was to evaluate the positive predictors of micrometastasis in central lymph nodes in patients with papillary thyroid cancer. Methods: This was a cross-sectional study. The study population was all known PTC patients who underwent total thyroidectomy and lymph node dissection of the central neck nodes based on the current indications. Confirmation of central lymph node involvement was performed by permanent smear after surgery. Data were analyzed using SPSS software version 22. A P-value below 0.05 was considered statistically significant. Results: There was no significant relationship between age, gender, family history of PTC, family history of thyroid disease, multinodularity, history of other thyroid diseases, involvement of two thyroid lobes, and tumor grade with central lymph node involvement (P>0.05). There was a significant relationship between the tumor pathology and size with central lymph node involvement (P<0.05). Moreover, logistic multivariate regression analysis showed that female gender, multinodularity, and tumor size had a significant relationship with the incidence of central lymph node involvement (P<0.05).   Conclusion: Female gender, multinodularity, and larger tumor size may be predictors of micrometastasis in central lymph nodes in patients with papillary thyroid cancer.

Abstract Background & Objective: The vaccine available to prevent Hepatitis B virus disease is ineffective in 5% of people due to the use of HBsAg as a weak immunogen. In the present study, PreS2/S fused to C18-27 peptide fragment as an effective antigen and is proposed as a promising vaccine candidate compared with the conventional vaccine prescribed in the vaccination program.
Methods: After the synthesis of PreS2/S genes and C18-27 peptide fragment in pET28a, the recombinant protein was confirmed by Western blotting. The efficacy of the PreS2/S-C18-27 protein was compared with the conventional vaccine injected into five groups of rats. Finally, the cytokine level of IF-r, IL-2, IL-4, IL-10, TNF-a, IgG1, and IgG2a were measured using the ELISA method.
Results: This study showed no significant difference between the recombinant vaccine group and PBS control group in the IF-r test, but there was a significant difference between groups testing IL-2 and IL-10. In addition, the group receiving the recombinant vaccine with CPG adjuvant at a dilution of 1/10 in the IgG total test on days 14 and 45 after the first injection showed a significant difference in comparison with other groups.
Conclusion: This study showed no statistically significant difference between the recombinant protein vaccine group and the conventional vaccine group. The Th1- mediated immune responses obtained from recombinant proteins with and without CPG performed better than conventional vaccines, possibly due to the functional deficiency of the available vaccines.

Abstract Background & Objective: A burn wound is sterile immediately after injury, but opportunistic bacteria colonize the wound within 48 to 72 hours after the burn, causing delayed or failed burn wound healing. In addition, the presence of multidrug-resistant (MDR) pathogens doubles the treatment problems. Lactobacillus plantarum (L. plantarum) is a well-known antibacterial and healing agent that could be used topically to treat burn wounds.  
Case Series Presentation: This clinical trial study (Case Series) was performed on 20 patients with deep second-degree burns. Patients had bilateral wounds; the wound on one side of the body was considered as control (treated with silver sulfadiazine) and the other side of the body as treatment (treated with bacteria-free supernatants (BFS) of L. plantarum). The wounds were evaluated by microbial assessments and assessments related to healing. Pseudomonas aeruginosa, Klebsiella pneumonia, and Staphylococcus aureus were isolated from 4 (22.2%), 0%, and 2 (11.1%) of wounds treated with L. plantarum on the fifth day of the treatment, respectively. Furthermore, 12 (66.7%) of wounds treated with L. plantarum were free from bacteria. The need for skin grafting was the same in both treatment and control groups, but graft rejection in the group treated with L. plantarum was (0%) (P=0.02).
Conclusion: Regarding eliminating or reducing infection and wound healing, bacteria-free supernatants of L. plantarum can be considered a possible topical treatment option in the case of second-degree burn wounds.

Abstract Background & Objective: Emerging evidence suggests that KRAS could play an important role in squamous cell carcinoma; however, its role in oral squamous cell carcinoma (OSCC) is largely unknown. The aim of the current study was to investigate the expression of KRAS, Ki-67, Cyclin D1, and Bcl2 in OSCC and their association with clinicopathological features. Methods: Forty paraffin blocks of retrospective histologically diagnosed cases of OSCC and 20 blocks of oral leukoplakia with epithelial dysplasia were obtained from two hospitals between 2018 and 2021. The paraffin-embedded tissue was analyzed for the expression of KRAS for oral epithelial dysplasia and OSCC, and ki-67, Cyclin D1, and bcl2 were analyzed only for OSCC. The results were correlated with each other and with different clinicopathological features and were statistically analyzed. Results: KRAS expression was significantly associated with histological tumor grade, tumor extent, presence of nodal and distant metastasis, pathological stage, and the presence of lymphovascular invasion (P=<0.001, 0.001, 0.001, 0.009, <0.001, and <0.001, respectively). The KRAS expression was positively correlated with the histological grade, tumor extent, nodal status, and the pathological stage (r=0.712, 0.649, 0.646, and 0.865, respectively). A positive correlation was also found with the expression of Bcl2, Cyclin D1, and Ki-67 (r=0.81, 0.723, and 0.698, respectively). The KRAS expression in oral epithelial dysplasia was significantly lower than that in OSCC (P=0.003).

Abstract Background & Objective: Invasive breast carcinoma of no special type (IBC-NST) is the most common type of breast cancer, which mainly causes axillary lymph-node metastasis (ALNM). Building on our previous research, we wanted to explore the optimal combination of AKT2, CD44v6, and MT1-MMP for the ALNM prediction.
Methods: The presence or absence of ALNM was used to separate 46 paraffin blocks containing IBC-NST primary tumors into two groups. Age, tumor grade, tumor size, receptor status (ER, PR, HER2, Ki-67, TOP2A), and test biomarker expression were evaluated. Biomarker expressions were assessed by IHC staining and categorized according to their respective cut-offs from our previous study, while other data were collected from archives. Data was gathered and analyzed using univariate, multivariate, and AUROC models.
Results: The expression of CD44v6 (OR: 12.77, 95% CI: 2.18-87.12, P=0.005) was identified as the independent variable for ALNM. Meanwhile, AKT2 expression (OR: 3.22, 95% CI: 0.36-22.41, P=0.237) and MT1-MMP expression (OR: 5.35, 95% CI: 0.83-34.54, P=0.078) did not demonstrate a statistically significant independent association in respect to ALNM. Combining AKT2 and MT1-MMP on CD44v6 increased overall accuracy by 4% compared to CD44v6 alone (AUROC 0.89 vs. 0.85).
Conclusion: The combined usage of AKT2, CD44v6, and MT1-MMP revealed no significant change compared to CD44v6 alone. Due to cost and practicality, we propose using CD44v6 as a biomarker predictor of ALNM in IBC-NST.

Abstract It is very rare for colorectal neoplasms to metastasize to the heart in the worldwide medical literature; only a single case of well-documented colorectal cancer metastasis to the left atrium was found. The case of a 66-year-old man is explained in this paper, who was suffering from metastatic adenocarcinoma of the colon that included the left atrium. In transthoracic and transesophageal echocardiography, a large multilobulated mass was present in the left atrium. An accidental pulmonary mass was also seen in a lung spiral CT scan. The cardiac mass was taken out, and a biopsy was obtained from the pulmonary mass. Adenocarcinoma was seen in histological assessment. Immunohistochemical staining was carried out to examine the expression of cytokeratin 7, cytokeratin 20, and caudal-related homeobox transcription factor 2 (CDX2) to determine the origin of the adenocarcinoma. In addition, the expression of these proteins was linked to the attributes of the patient and tumor. Post-surgery transesophageal echocardiography showed normal left ventricle and right ventricle function with no evidence of left atrium mass. Therefore, we suggest that asymptomatic cancer patients with a history of colorectal cancer and who have developed cardiac symptoms should be immediately examined for potential cardiac metastasis.

Abstract Crescentic glomerulonephritis (GN) is a feature of severe glomerular injury. Anti-GBM disease, immune-complex mediated glomerulonephritis, and ANCA-associated vasculitis are the main causes of crescentic GN. Alport syndrome is a progressive form of hereditary nephritis presenting with hematuria and progression to proteinuria and renal failure. Herein we present a 16-year-old male with rapidly progressive glomerulonephritis syndrome, sensory-neural hearing loss, and a family history of hematuria and proteinuria in his mother and aunt. Light microscopic examination shows cellular crescent in glomeruli. In an electron microscopy study, GBM changes compatible with Alport syndrome were identified. Alport syndrome rarely can be presented as crescentic GN. Electron microscopy is necessary for the diagnosis of this type of pauci-immune crescentic glomerulonephritis.