Background: Human epidermal growth factor 2 (HER2) is known to be an important prognostic factor in breast cancer. Numerous studies have shown HER2+ breast cancers have reduced overall survival and recovery time, as well as the efficacy of anti-HER2 therapies along with chemotherapy in improving disease outcomes. For this reason, it is recommended that all patients with breast cancer should be evaluated for HER2 status. This study aimed to assess the HER2 gene amplification by the CISH method in evaluating the HER2 status in patients with immunohistochemistry (IHC) 2+ (equivocal) results. Methods: This retrospective cross-sectional study examined HER2 status based on the Chromogenic in situ hybridization (CISH) method in 280 breast carcinoma samples with an initial 2+ (equivocal) score in IHC. The relationship between HER2 amplification and hormone receptors (estrogen and progesterone) and Ki67 level was also investigated. Results: In sixty samples (21. 4%), the HER2 gene was amplified based on the CISH method. The majority (215, 76. 8%) of the samples were negative and 5 (1. 8%) samples were indeterminate. No significant relationship was observed between HER2 amplification, estrogen receptor (p=0. 143), and Ki-67 protein level (p=0. 977). There was a significant inverse relationship between HER2 amplification and progesterone receptor positivity (p=0. 007). Conclusion: These results demonstrate that CISH is a helpful method to assess HER2 status in equivocal breast cancer and is positive (amplified) in about 21. 4% of them.

Background: The incidence rate of gastric cancer in Western countries has shown a remarkable decline in recent years although it is still the almost common cancer between men in Iran. The proto-oncogene MYC, located at 8q24.1, regulates almost 15% of human genes and is activated in 20% of all tumors. MYC amplification and overexpression of its protein product are observed in 15-30% of gastric neoplasia. The objective of this study was to find the preference of CISH or IHC in the diagnosis and prognosis of gastric cancer.Methods: In this cross-sectional investigation, 102 paraffin blocks samples of Iranian patients with gastric cancers were studied. All the patients had undergone primary surgical resection at the Cancer Institute Hospital, Tehran University of Medical Sciences from 1987 to 1993. CISH and IHC techniques were applied to the samples. CISH was carried out on 3-mm-thick tissue sections and with a ZytoDot CISH Implementation Kit (ZytoVision GmbH, Germany). IHC was down using the HRP method with the monoclonal antibody. A universal peroxidase-conjugated secondary antibody kit was used for the detection system. All samples were gastric adenocarcinoma and were selected randomly.Results: Our data revealed that both diffuse and intestinal types of gastric cancer occurred significantly in men more than women. Our results showed an indication of some correlation between grades and CISH results, although the difference was not significant.Our data also showed that CISH+patients (43.1%) were more frequent in comparison with IHC+patients (14.7%). There was a correlation between CISH and IHC. This result revealed that there was a significant difference between grades and IHC. There was also no statistically significant difference between CISH amplification in diffuse and intestinal types.Conclusion: Our conclusion is that for the treatment, management of stomach cancer, and monitoring of progress and prognosis of the tumor that is almost important for patients and clinicians, CISH test is a better and feasible to IHC test, with regards to sensitivity and specificity.

Background: Gastric cancer has a high mortality rate and often has a poor treatment outcome. The HER2/neu gene target therapy has been known as a potential way for treatment.Objectives: The goal of our study was assessment the relation between chromogenic in situ hybridization (CISH) and immunohistochemistry (IHC) methods in determining the best diagnostic method for gastric cancer.Methods: In this historical cohort study, 50 gastric cancer samples were analyzed by CISH and IHC. The relation between clinicalpathological parameters of HER2/neu was also analyzed. Alive patients were followed from 2009 through 2012 for the main outcomes (mortality). The results of these two methods, in terms of sex, age, tumor size, grading, staging, tumor location, metaplasia, presence of necrosis and ulceration, vascular invasion, the TNM system, mucin or signet producing adenocarcinoma cells and patient survival rates were compared.Results: There was no significant difference between IHC and CISH regarding the sex, age, tumor size, grading, staging, tumor location, metaplasia, presence of necrosis and ulceration, vascular invasion, the TNM system, mucin or signet producing adenocarcinoma cells and patient survival rates. Comparison of TNMscores by these two methods showed no significant relationship between IHC and staging, but a statistically significant difference between CISH and different N staging, (P<0.05) was assessed.Conclusions: Comparison between IHC and CISH showed the only significant relationship between CISH and different N staging.Therefore, low amplified CISH was a better diagnostic method for gastric cancer, compared to low expression in IHC.

Background: Diffuse large B-cell lymphoma (DLBCL) is the most common type of lymphoma. There are various types of DLBCL including immunoblastic and centroblastic. Epstein-Barr virus (EBV) is a member of Herpes virus family found in all human populations inducing different lymphoproliferative disorders. The role of EBV in the development of DLBCL is known. Multiple laboratory methods are available for detecting EBV. This study was conducted to determine the correlation of EBV with DLBCL in samples referred to pathology ward in Shariati and Sina Hospitals by chromogenic in situ hybridization (CISH) method.Methods: In this case/control study, pathological specimens of 50 patients with DLBCL as well as 50 reactive lymph nodes and tonsils (control group) were collected from archives of Shariati and Sina Hospitals and were evaluated for EBV encoded RNA (EBER) expression based on CISH method. A peptide nucleic acid (PNA) EBV probe (Dakocytomatin) was used while all the processes were done in RNAase-free conditions using RNAase-free water, sterile gloves and samplers.Results: Out of fifty specimens in the case group, eight were positive for EBER in comparison with two in the control group (P=0.046). No statistically significant difference was observed between intranodal or extranodal samples (P=0.736) or between males and females (P=0.0746).Conclusion: Our study showed that EBV positivity for EBER in patient with DLBCL could be determined more effectively by CISH method than immunohistochemistry (IHC). Comparative analysis between CISH, PCR and IHC methods is recommended.

Background & Objective: The her2 amplification plays an important role in breast cancer management. Therefore, there is a need for using supplementary molecular methods in IHC equivocal cases. Present study has been conducted to determine the effects of clinicopathological variables on her2 gene amplification by chromogenic in situ hybridization (CISH) in IHC Her2 (2+) breast cancer individuals. Methods: A cross-sectional study was conducted in Zaferanyeh Laboratory collaborated with Shahid Beheshti University of Medical Sciences (Tehran-Iran; 2015-2018). All pathological data related invasive breast cancer patients with equivocal IHC results were included. CISH method was performed as a supplementary technique. The associations between histopathologic variables, status of Ki-67 index, progesterone and estrogen receptors (PR & ER) with her2 amplification by CISH were investigated and analyzed. The level of significance was considered as P-value < 0. 05. Results: Totally, 239 patients with mean age of 53. 2 years were studied. CISH identified her2 gene amplification in 51 subjects (21. 3%). The type of tumor (invasive ductal carcinoma), the tumor grade, and the value of Ki-67 index were directly correlated with her2 amplification. Significant negative associations were also observed between CISH results and ER and PR expression. Conclusion: As her2 gene amplification was identified in 21. 3% of invasive breast cancer patients with equivocal IHC results, it is supposed that applying CISH method may consider as a potentially valuable supplementary method. Results have also shown that higher grades of tumor, invasive ductal carcinoma, absences of hormone receptors and high Ki-67 index significantly correlated with the her2 amplification.

Background: The incidence rate of gastric cancer in western countries has shown a remarkable decline in the recent years while it is still the most common cancer among males in Iran. The proto-oncogene MYC, located at 8q24. 1, regulates almost 15% of human genes and is activated in 20% of all tumors. The amplification of MYC and overexpression of its protein product are observed in 15-30% of gastric neoplasias. Objectives: The objective of this study was to find the preferences of Chromogenic In Situ Hybridization (CISH) and Immunohistochemistry (IHC) in diagnosis and prognosis of gastric cancer. Patients and Methods: We studied 102 samples of gastric cancer in Iran and all the patients had undergone primary surgical resection at the Cancer Institute Hospital, Tehran University of Medical Sciences. The CISH and IHC techniques were applied for all our samples. All of the samples had adenocarcinoma gastric cancer and were selected randomly. Also, the type of study was cross sectional. The sample size was 100 patients. Results: Our data revealed that both diffuse and intestinal types of gastric cancer occurred significantly more in males than females. Our results showed that there was an indication of some correlation between grades and CISH, although the difference was not significant. Our data also showed that CISH positive patients (43%) were more frequent compared to IHC positive patients (14. 7%). There was a correlation between CISH and IHC. These results revealed that there was a significant difference between grades and IHC. There was also no statistical difference between CISH amplification in diffuse and intestinal types. Conclusions: From the results, it could be concluded that for administration of the treatment of stomach cancer, and progress and prognosis of tumor, which is important for patients and clinicians, the CISH is a better and more feasible test than IHC, in regards to sensitivity and specificity.

Background: Chromogenic insitu hybridization (CISH) and immunohistochemistry analysis for p57 are ancillary studies discriminating partial hydatidiform mole (PHM), complete hydatidiform mole (CHM), and non-molar hydropic abortion (HA). Objective: It aimed to study CISH with a probe to chromosome 17 (CISH17) and chromosome 2 (CISH2) discriminating chromosomal ploidy of PHM, CHM, and HA; in addition, their surrogacy value in the evaluation of triploid and diploid in product of conception specimens (POCs) was evaluated. Materials and Methods: 44 statistically significant POCs were selected retrospectively. The Kappa agreement coefficients, sensitivity, specificity, and accuracy with 95% confidence interval (95% CI) were reported. Results: PHM, CHM, and HA were diagnosed to be 23, 17, and 3 cases based on both CISH2 and CISH17 resulting in their complete discrimination between PHM and HA (23 vs. 3). The Kappa agreement coefficient was 95.4% (p < 0.001) when diagnosing the PHM (23), CHM (20), and HA (1). In addition, the accuracy, sensitivity, and specificity were 95.26% (95% CI: 84.25-99.38), 100% (95% CI: 85.18-100), and 95% (95% CI: 76.18-99.88), respectively. The power analysis on CISH2 and CISH17 tests discriminating between triploid and diploid in POCs was estimated to be 100%. Conclusion: Based on the current finding, CISH2 and CISH17 enjoyed perfect agreement in diagnosing chromosomal ploidy; in addition, their absolute power discriminating between triploid and diploid revealed that they could be used as surrogate markers for ploidy. Prospective studies on fresh specimens are suggested comparing the CISH method's accuracy with flow cytometry karyotyping and fluorescence in situ hybridization. This article has been extracted from M.D. Thesis. (Tara Taheri)

Background and objective: The assessment of human epidermal growth factor receptor 2 (HER2) status has become of great importance in the diagnosis of breast cancer. The aim of this study was to investigate the diagnostic value of quantitative Polymerase Chain Reaction (qPCR) and Chromogenic In Situ Hybridization (CISH) to assess HER2 status of biopsy specimens.Methods: To elucidate the status of HER2 gene amplification, biopsies of breast carcinoma from 120 patients with 2+IHC status were analyzed by qPCR and CISH.Results: The results of the two experiments were compared, and it was depicted that the concordance rate between CISH and qPCR assays was 88.1%.The quantification of HER2 gene with CISH and qPCR showed that there was a significant correlation (p value=0.0001 and r=0.808).Conclusion: The results of this research support the idea that qPCR is a precise and reproducible technique, which can be employed as a supplementary method to evaluate HER2 status.

Introduction: Cisplatin is a strong anticancer medicine, but its use is limited due to the potential nephrotoxicity induction. Objectives: The present study seeks to determine the impact of Hypericum hydroalcoholic extract on cisplatin-induced nephrotoxicity. Materials and Methods: Thirty-two male rats were assigned to groups 1 to 4. Group 1, control (Cont),treated by saline (IP). Group 2, Cis,cisplatin [intraperitoneal (IP), 7. 5 mg/kg]. Group 3, CISH,cisplatin + Hypericum (70 mg/kg, IP, for one week). Group 4, HCis,first treated with Hypericum for a week, followed by cisplatin. Renal tissue and blood samples were obtained a week after cisplatin injection for tissue assay and biochemical analysis. Kidney tissue damage score (KTDS), plasma creatinine (Cr), blood urea nitrogen (BUN), serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) were measured. Results: Kidney weight showed significant differences between the treated groups and the Cont group (P<0. 001). Serum BUN, Cr, SGOT, and SGPT increased significantly in Cont (P<0. 01). BUN decreased in CISH and HCis groups compared to Cis group, although there was no significant difference. Serum Cr, SGOT, and SGPT decreased significantly in CISH and HCis groups compared to the Cis group (P<0. 05). MDA and KTDS increased in the Cis group and decreased significantly in the CISH and HCis groups compared to the Cis group (P<0. 05). Serum SOD and CAT decreased significantly in Cis compared to Cont (P<0. 05) and increased in CISH and HCis groups compared to Cis. There was no significant difference between the CISH and HCis groups in any of the measured parameters. Conclusion: This study reveals that pretreatment with Hypericum extract or its concomitant administration with cisplatin can moderate the side-effects of cisplatin, improve renal function and decrease lipid peroxidation, renal toxicity and the KTDS.

Introduction & Objective: The epidermal growth factor receptor gene (HER-2) plays an important role in cell transformation and tumorgenesis. The presence of HER-2 gene generally predicts a good response to anthracycline-based agents and therapies that specifically target HER-2 (Herceptin).Materials & Methods: Comparison between IHC and CISH methods for assessment of HER-2 status was performed on 38 women suffering from invasive ductal carcinoma. Data analysis was performed by pair wise grouped statistical method.Results: There was a good correlation between IHC and CISH methods for assessment of HER-2 status in 92.1% of cases (Kappa=0.938, P<0.001).Conclusions: CISH method is recommended as a complementary method in the assessment of HER-2 status in invasive ductal carcinoma of breast of those having weakly positive HER-2.