فهرست مطالب

Pathology - Volume:13 Issue:4, 2018
  • Volume:13 Issue:4, 2018
  • تاریخ انتشار: 1397/09/03
  • تعداد عناوین: 15
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  • Farid Kosari *, Fatemeh Ghaffari Pages 390-396
    Background and Objective
    Hodgkin's lymphoma is a potentially curable hematologic malignancy with difficulty in its diagnosis especially in atypical cases even in expert hands. Today, immunohistochemistry plays a significant role in the diagnosis of it especially applying the anti-CD15 and anti-CD30 antibodies. The negativity of CD15 can be reduced by antigen retrieval for methods. In this study, the effect of autoclave was compared with microwave as heating sources of antigen retrieval in immunohistochemical staining.
    Methods
    Sections prepared from 50 formalin-fixed paraffin-embedded tissue blocks of Classic Hodgkin's lymphomas stained for CD15 and CD30 using autoclave and microwave, were randomly and blindly reviewed by an expert hematopathologist, mostly focusing on Reed-Sternberg cells; the intensities were scored from 0 to +4 and analyzed by SPSS software.
    Results
    Fifty eight percent of patients were male. The mean age was 32 years (range: 7 to 77). Nodular sclerosis was the most prevalent subtype. CD15 positivity in microwave treatment was 92% compared to 50% in autoclave. Negative CD30 decreased from 20% in autoclave to 2% in microwave. Intensity of staining in both markers was at least +1 greater in microwave treatment. No background staining was seen in microwave method.
    Conclusion
    There was bimodal age distribution in Hodgkin's lymphoma patients with the predominant male and Nodular Sclerosis as the most common type. Comparing autoclave and microwave, higher rate of the positivity was detected using microwave treatment, especially in CD15 staining. Improvement in staining intensity was also noticeable in both markers. There was no background or non-specific staining in microwave method. No disturbance of cells or nuclear morphology was also reported.
    Keywords: Hodgkin Disease, Immunohistochemistry, Microwaves, CD15 antigen, CD30 antigen
  • Seyyede Fatemeh Shams, Hossein Ayatollahi, Mohammad hadi Sadeghian, Monavar Afzal Aghaee, Sepideh Shakeri, Ehsan Yazdandoust, Maryam Sheikhi *, Nafiseh Amini, Samane Bakhshi, Afsane Bahrami Pages 397-402
    Background and Objective
    Janus kinase 2 (JAK2) and Myeloproliferative Leukemia (MPL) mutations are confirmatory indicators for Myeloproliferative Neoplasm (MPN). The current study was performed to determine the frequency of MPL mutation in MPN patients without JAK2 mutation, in order to assign MPL mutation frequency in North-East of Iran.
    Methods
    Total of 105 negative JAK2 cases including 5 Myeloproliferative Disorders (MPD), 15 Polycytemia Vera (PV) and 15 Essential Thrombocytosis (ET) who referred to Qaem Medical Center were assigned to this study. ARMS-PCR was carried out for measuring MPL mutations.
    Results
    A significant difference was observed between MPL mutant and non-mutant groups from overview of MPL mutation (P=0.00001). From the total studied population, 14.28% were ET cases and 4.71% of them had splenomegaly. About 66.66% had thrombocytosis and 33.33% of all the individuals had leukocytosis according to WHO criteria, and 4.76% of non-MPL mutant individuals had splenomegaly (P=1).
    This mutation was reported in 4-6% of ET and PMF individuals. In this research, 4.76 % of studied individuals had MPL (W515L/K) mutation, which were diagnosed with ET.
    Conclusion
    Generally, the presence of JAK2 and MPL mutations are the most important criteria for MPN diagnosis. The obtained frequency of MPL mutation was similar to previous studies. Despite the high frequency of JAK2 and Philadelphia abnormality, MPL mutation was rare in myeloprolifrative disorders. Further studies are suggested to investigate its prognostic effects for these diseases.
    Keywords: MPL (W515K-L), JAK2 (V617F), Myeloproliferative neoplasm
  • Faria Hasanzadeh Haghighi, Ehsan Aryan, Mohammad Derakhshan, Aida Gholoobi, Zahra Meshkat* Pages 403-407
    Background & objective
    Tuberculosis (TB) remains a major cause of death around the world. Bacillus Calmette Guérin (BCG) is the only vaccine used in TB prevention that has a protective effect in children, but its effectiveness declines in adults. Design and development of new vaccines is the most effective way against TB.
    The aim of this study was to design and construct a DNA vaccine encoding mtb32C and mpt51 fusion genes of Mycobacterium tuberculosis.
    Methods
    First, mpt51 fragment was amplified by PCR method. The pcDNA3.1+/mtb32C plasmid was transformed into E. coli JM109 and then extracted. The mpt51 gene and pcDNA3.1+/mtb32C plasmid were both digested with EcoRI and BamHI restriction enzymes followed by ligation of mpt51 fragment into the digested vector. The recombinant plasmid containing mtb32C and mpt51 was subsequently transformed into competent E. coli TOP10 strain. The clones were confirmed by colony-PCR, restriction enzyme digestion and sequencing.
    Results
    Using agarose gel electrophoresis, a 926 bp fragment corresponded to mpt51 was observed. Digestion of the vector pcDNa3.1+/mtb32C and mpt51 gene was confirmed by electrophoresis. Then, the pcDNA3.1+/mtb32C plasmid was extracted. Sequencing results confirmed the accuracy of the desired plasmid.
    Conclusion
    In this study, we constructed a cloning vector encoding Mtb32C/Mpt51 gene of Mycobacterium tuberculosis. The eukaryotic expression of this vector can be confirmed in future studies. It can be considered as a DNA vaccine in animal models later. Successful cloning provides a basis for the development of new DNA vaccines against tuberculosis.
    Keywords: Mycobacterium tuberculosis, antigens, genetic vectors, Cloning
  • Narjes Seid Alian, Parvin khodarahmi *, Vahid Naseh Pages 408-414
    Background
    Cadmium is a potent toxicant and carcinogenic in human and experimental animals. The evidence indicates that cadmium induces aberrant gene expression, inhibition of DNA damage repair, and apoptosis. In this study, we investigated the effects of IP (intraperitoneal) injection of cadmium on mRNA levels expression of Bcl-2 and Bax genes in rat small intestine.
    Methods
    28 male Wistar rats weighing 200 to 250 grams were randomly distributed into 4 groups. Group 1 received saline while the animals in groups 2-4 were injected cadmium at doses of 1, 2 and 4 mg/kg of cadmium for 15 successive days. One day after the last injection, the small intestine was dissected and the mRNA levels expression of Bax and Bcl-2 genes was evaluated using Real Time PCR technique.
    Results
    Cadmium increased the mRNA levels of Bax gene compared to the control group at 2 and 4 mg/kg (p < 0.01) in small intestine of rats. The mRNA levels of Bcl-2 gene decreased significantly compared to the control group at 1, 2 and 4 mg/kg (p < 0.001) in small intestine of rats.
    Conclusion
    These results showed Cadmium exposure induced cell apoptosis by increasing Bax/Bcl-2 ratio expression.
    Keywords: Cadmium, Bax, Bcl-2, Small Intestine
  • Amir Hossein Jafarian, Khatoone Mirshekar, Sare Etemad, Masoumeh Jafaripour, Mansoore Darijani, Maryam Sheikhi, Hossein Ayatollahi *, Sepideh Shakeri, Seyyede Fatemeh Shams, Saeed Davari Pages 415-421
    Background and Objective
    BRAF mutations were studied in various populations for prostate carcinoma (PC); however, mutations in BRAF gene are unusual compared to KRAS. Oncogenic activating of BRAF mutations were studied lately in almost 0%-10% of prostate cancer cases.
    Methods
    In this retrospective study, we gathered 100 formalin-fixed paraffin-embedded samples of prostate adenocarcinoma. A hundred archived samples of adjacent benign prostatic hyperplasia were chosen as normal control. This study was done in pathology laboratory of Qaem Hospital during 2013-2015.
    Results
    Total number of 200 PC and normal cases was investigated for BRAF V600E mutation. The BRAF V600E mutation was found in only 4 patients but it was not detected in normal cases. There were no significant differences between patient and control groups for this mutation (P>0.99). The frequency of BRAF V600E mutation was not significant in different age groups (P>0.285); the most frequency was related to the age range of 71-80. No significant difference was observed between tumor grade and BRAF mutation (P=0.21).
    Conclusion
    According to our findings, BRAF gene mutations did not play essential role in PC. Therefore, anti-BRAF (V600E) could not be considered as a proper target for therapy.
    Keywords: BRAF mutation, Prostate adenocarcinoma, Gleason score
  • Navid Bazzaz, Nazila Nouraee Nouraee, Ali Zare, Mirzaie *, Seyed Javad Mowla Mowla, Maryam Shahali, Mohammad Vasei Pages 422-428
    Background and Objective
    Wilms’ tumor (WT) is the most common genitourinary tract tumor in children. MicroRNAs (miRNAs) are small non-coding RNAs; their role in the pathogenesis of many types of human cancers has been identified. We aimed to evaluate the expression of miR-21, a well-known oncomir, in WT tissue samples which is a very common urinary tract malignancy in children.
    Methods
    We performed chromogenic in situ hybridization (CISH) to detect the sub-cellular localization of miR-21 in 25 formalin-fixed, paraffin-embedded (FFPE) samples of WT. We also evaluated miR-21 expression in 24 of these blocks and 6 normal kidneys as controls using quantitative real-time PCR technique.
    Results
    While our real-time PCR analysis showed miR-21 significant overexpression in 4 tumors compared to the normal kidney samples, we could not detect significant ISH signal in any of these samples.
    Conclusion
    Low expression of miR-21 in WT might pinpoint the weak involvement of this miRNA in the pathogenesis of this cancer.
    Keywords: Wilms’ tumor, miR-21, CISH, Real-time PCR
  • Amir Tajbakhsh, Faezeh Ghasemi, Seyedeh Zohre Mirbagheri, Mastoureh Momen Heravi, Mehdi Rezaee, Zahra Meshkat* Pages 429-437
    Background and Objectives
    The incidence of rifampin-resistant strains of Mycobacterium tuberculosis has attracted more attention than the tuberculosis infection due to laborious treatment and control. Recognizing the Mycobacterium tuberculosis genotypes involving in drug resistance via multiplex PCR, a simple and rapid genotyping method, is an emergency for better treatment and control of tuberculosis. This study was designed to specify the frequency of rifampin-resistant strains of Mycobacterium tuberculosis isolated from patients by multiplex allele-specific Polymerase Chain Reaction assay (MAS-PCR).
    Methods
    In this study, 88 Mycobacterium tuberculosis positive samples were included from Qaem Hospital, Mashhad. MAS-PCR was used to detect the rifampin resistance associated mutations in rpoB gene.
    Results
    Mutations in three codons of rpoB gene causing rifampin resistance were detected in 51 isolates (58.96%). The detected mutations in codons 531, 526, and 516 were 55.68%, 38.63%, and 13.63%, respectively. The simultaneous mutations were detected in 11 isolates (12.50%) in codons 531, 526 and 516, in 21 isolates (23.86%) in codons 531 and 526, and in one isolate (1.13%) in codons 526 and 516.
    Conclusion
    According to the results of this study, the frequency of rifampin-resistant strains of Mycobacterium tuberculosis isolated from Khorasan province patients (North-East of Iran) was high. The developed MAS-PCR assay can be used for rapid detection in clinical diagnostic laboratories in areas with high prevalence of multidrug-resistant Mycobacterium tuberculosis strains. In this respect, MAS-PCR is simple, rapid, and highly sensitive method for drug susceptibility tests for detecting multidrug-resistant Mycobacterium tuberculosis.
    Keywords: Multiplex PCR, rpoB gene, rifampin resistance, Mycobacterium tuberculosis
  • Freidoon Solhjoo *, Akbar Safaie, Ahmad Monabati, Maral Mokhtari, Moeinadin Safavi Pages 438-446
    Background and Objective
    Identification of cytogenetic and molecular changes plays an important role in acute myeloid leukemia (AML) patients. Thus, they are used in classification, prognosis and treatment of the disease. The CD123 expression and FLT3 gene mutations are also the variations that may assist in prognosis and treatment of patients with AML.
    Methods
    This study was performed on 76 patients as new cases of AML. The correlation between CD123 immunohistochemical (IHC) expression and FLT3 gene mutations with each other as well as morphological, immunophenotypical and cytogenetic factors was studied.
    Results
    The results represented the CD123 IHC expression in 55.3% and FLT3 gene mutations in 28.9% of cases. We found that 81.3% of patients who had FLT3/ITD gene mutations revealed IHC of CD123 expression (P=0.019). The CD123 expression against FLT3 was also correlated with monocytic differentiation in bone marrow blasts (P=0.031). There were significant correlations between IHC expression of CD123 and FLT3/ITD mutations with a high percentage of aspirated bone marrow blasts (P=0.01 and P=0.006, respectively) as well as the lack of CD34 expression in bone marrow blasts (P=0.007 and P=0.021, respectively).
    Conclusion
    The CD123 IHC positive AMLs were correlated with certain pathologic features, some of which can be similar with correlations of background mutation of FLT3/ITD; According to the negative predictive value (NPV), 88.2% of CD123 IHC showed FLT3 gene mutation. In addition to its use in targeted therapy, it could be a marker to decide what molecular tests to use in the next steps.
    Keywords: Acute myeloid leukemia, FMS-like tyrosine kinase 3, CD123, Karyotype
  • Shadi Hosseini, Farkhondeh Behjati, Maryam Rahimi, Nazanin Taheri, Hamid Reza Khorram Khorshid, Fatemehte Aghakhani Moghaddam, Saghar Ghasemi Frouzabadi, Masoud Karimlou, Fereidoon Sirati, Elahe Keyhani* Pages 447-453
    Background and Objective
    The PI3K/AKT/mTOR pathway is known to play an important role in regulating angiogenesis both in normal and breast cancer (BC) tissues. PIK3CA amplification was reported in various malignancies, including approximately 10% of BC cases. The aim of this study was to identify the frequency of PIK3CA amplification in Iranian female patients suffering from BC. Additionally, possible association between PIK3CA amplification and P110α expression with microvascular density (MVD) was examined.
    Methods
    DNA samples were extracted from paraffin embedded tumor tissue blocks and copy number changes were evaluated by MLPA Technique. The results were analyzed by coffalyzer software. The tissue expression of P110α and CD34 was assessed using immunohistochemistry.
    Results
    Ten out of 40 samples (17.5%) showed amplification in PIK3CA gene and 22 out of 40 samples (55%) showed overexpression in P110α. For CD34, from 40 samples, 20 (50%), 15 (37.5%) and 5 (12.5%) had scores 1+, 2+ and 3+, respectively.
    Conclusion
    No significant association was detected between gain of PIK3CA copy number and P110α or CD34 tissue expression.
    Keywords: PIK3CA, Amplification, P110?, MVD, Breast cancer
  • Seyedeh Mehrnaz Kouhbanani nejad, Farzaneh Armin, Shahriar dabiri, Ali Derakhshani, Maryam Iranpour, Alireza Farsinejad* Pages 454-460
    Background and Objective
    In recent years, due to increasing number of patients with non-healing skin ulcers, skin substitutes have been used. Skin substitutes contain living cells causing faster and more effective wound healing. Therefore, research on the use of autologous and allogeneic cells such as fibroblasts in skin substitutes has attracted attentions. However, there are discrepancies in the immune responses to allogeneic fibroblasts. Therefore, we aimed to review the immune responses to allogeneic fibroblasts.
    Methods
    Donor fibroblasts were isolated from the skin of three rats. Nine recipient rats which were subcutaneously injected with three different regimens, were divided into three groups: Group 1; phosphate buffered saline (PBS) without cells (control), group 2: allogeneic fibroblasts of one animal source suspended in phosphate buffered saline, and group 3; phosphate buffered saline containing mixed allogeneic fibroblasts of three animal sources. The skin samples were biopsied at 1, 3 and 7 days after injection and studied histopathologically. Results and Conclusion: No signs of redness and edema were observed in the injection sites. In pathology examination, changes such as vasculitis, eosinophils and lymphocytes accumulation around fibroblasts, fibroblast apoptosis and transplant rejection at the injection site were not observed in either group.
    Subcutaneous injection of allogeneic fibroblasts in rats can be introduced as a promising approach for wound healing as they do not stimulate the immune system.
    Keywords: Fibroblast, Allogeneic, Immune system, Wound healing
  • Noushin Pouryazdanpanah, Shahriar Dabiri, Ali Derakhshani, Reza Vahidi, Alireza Farsinejad * Pages 461-466
    Background and Objectives
    The mesenchymal stem cells derived from peripheral blood (PB) have been recognized as a promising source for allogeneic cell therapy. The aim of this study was to investigate the isolation, growth and differentiation ability of peripheral blood-isolated mesenchymal stem cells.
    Methods
    The mononuclear cells were purified from fresh peripheral blood using density gradient centrifugation then cultured in a suitable medium, expanded and characterized. In the following, these cells were cultured in specific adipogenic and osteogenic differentiation media. Results and Conclusion: In spite of the absence of any stimulating factor, the cells adhered to the flasks and developed a rather homogeneous, spindle-shaped morphology after consecutive passages. The cells were confirmed to have mesenchymal phenotype by expression of specific markers (CD90, CD105, and CD73) and absence of CD45 marker, which is specific for hematopoietic stem cells. They could differentiate into lineage-specific committed cells (osteoblasts and adipocytes).
    According to the findings, the conventional, labour-intensive and time-consuming approaches are not necessary to obtain an optimal number of cells from peripheral blood. This relatively accessible and minimally invasive source of stem cells may open a new era for practical exploitation in regenerative medicine.
    Keywords: Peripheral blood, Mesenchymal stem cells, differentiation, Regenerative medicine
  • Fatemeh Nili, Azadeh Sedighi moghadam pour, Hedieh Moradi Tabriz, Parisa Sedighi Moghadam Pour, Hana Saffar* Pages 467-470
    Peripheral primitive neuroectodermal tumor (pPNET) is a highly aggressive small round cell tumor belonging to PNET/Ewing sarcoma family. Ovarian tumors composed of primitive neuroectodermal elements are extremely rare.
    Herein we reported two cases of peripheral primitive neuroectodermal tumors of ovary in two patients with different clinical presentations. Definite diagnoses were made based on the histomorphology and immunohistochemistry
    results.
    With respect to different clinical behaviors, treatment modalities and prognosis of peripheral primitive neuroectodermal tumors compared to other known ovarian neoplasms, it is essential to consider this entity as a differential diagnosis in ovarian tumors especially in young patients.
    Keywords: Neuroectodermal Tumor pPNET, Ovary
  • Shilpa Bairwa *, Bhawna Sethi, Pawan Singh, Ashok Sangwaiya, Shivani Kalhan Pages 471-473
    A choriostoma is an aggregate of microscopically normal cells or tissues which occurs in an aberrant location. It follows a benign course, rarely seen in head and neck region. A choriostoma of the palatine tonsil is very rare; less than 10 cases were reported till date. A 11-year-old male referred to ENT OPD with chronic tonsillitis and underwent tonsillectomy. The histopathological examination revealed the unexpected presence of cartilage and bone in both tonsils.
    Keywords: bone, cartilage, choriostoma, tonsil
  • Gireesha Rawal, Sufian Zaheer *, Amit Yadav, Indrani Dhawan Pages 474-478
    Malignant fibrous histiocytoma (MFH) is one of the most common types of soft tissue sarcomas in adults. Distant metastases are developed in 30–40% of patients with MFH, with the most common site being the lung. However, metachronous MFH has not been reported previously in literature. This report describes a case of a 30-year-old man, who had two metachronous thigh tumors, both of which were confirmed to be MFH on histopathology and immunohistochemistry evaluations. A contemporary multidisciplinary approach to therapy including surgery, radiation and chemotherapy was advocated. Two primary sites of MFH raised the possibility of a genetic abnormality that could predispose such a patient to develop multiple primary sites of the same tumor.
    Keywords: Metachronous, Malignant fibrous histiocytoma
  • sora yaari *, Viroj Wiwanitkit Page 479
    Dear Editor, erythrocyte sedimentation rate (ESR) is a useful basic clinical pathology laboratory investigation. It can be helpful in diagnosis and follow-up of several diseases. At present, a new automated method with proven reliability is available for ESR test (1). Here, the authors report on observation on a laboratory experiment to test the effect of zinc nanoparticles on ESR results. The total of 100 blood samples was used in the experiment. Each sample was divided into two parts. One part was directly measured for ESR and the other part was added by 1 droplet of zinc nanoparticles solution then exposed to ESR measurement. All ESR measurements were done using the same automated ESR analyzer; MicroSed SR-system, in the same ISO15189 accredited clinical laboratory at the same time, place and condition. The ESR results showed a difference in ESR values between two groups. The ESR values for the groups with and without zinc nanoparticles were equal to 25.6 + 7.2 and 10.7 + 3.4 mm/hr, respectively. Therefore, zinc nanoparticles can interrupt the ESR test conducted by automated analyzer. This observation was similar to the recent report which showed nanoparticles can alter the result of lipid profile test (2). Therefore, due to the widely use of nanoparticle substances, practitioners must consider the effect of nanoparticles interference in the interpretation of ESR results.
    Keywords: Zinc, Nanoparticle, erythrocyte sedimentation rate