Immunohistochemical Evaluation of C4d and C3d Markers in Bullous Pemphigoid as a Substitute for Direct Immunofluorescence Technique.

Background: There are several methods for the diagnosis of autoimmune bullous disease. Direct immunofluorescent (DIF) testing is an important diagnostic method in the diagnosis of immunobullous disease but requires skilled pathologist, fresh tissue and well-equipped laboratory to perform the procedure. The immunohistochemistry analysis of C4d and C3d is easily compared with other methods. This study was conducted to assess the value of immunohistochemistry (IHC) analysis for expressions of C3d and C4d in the diagnosis of bullous pemphigoid (BP). Aims and Objectives: This study was conducted to assess the value of immunohistochemistry (IHC) analysis for expressions of C3d and C4d in the diagnosis of bullous pemphigoid (BP). Materials and. Method: We applied C4d and C3d immunohistochemistry on formalin-fixed, paraffin-embedded tissue on 30 cases of bullous pemphigoid that was confirmed by direct immunofluorescence (DIF) evaluation as well as 16 cases in control group (12 cases of herpetiform dermatitis, 3 cases of linear IgA dermatosis and 3 cases of bullous lichen planus). Results: Mean and SD of age were 68.13 ± 14.00, female to male ratio was 1:3. In cases where both C3d and C4d staining were positive, the intensity of C3d staining was higher than C4d. Twenty-two cases showed C4d-positive staining in IHC study, such that in seven cases focal staining and in 15 cases diffuse staining were observed. Also 26 cases showed C3d-positive staining in IHC study such that in four cases focal staining and in 22 cases diffuse staining were observed. In cases with C3d-positive staining, there were 21 cases of deposition only on the bullous floor, one case on the bullous roof and four cases on the bullous roof and floor. In cases with C4d-positive staining, there were 17 cases of deposition on the bullous floor, two cases only on the bullous roof and three cases on the roof and floor. All control cases were negative for C3d and C4d staining in the dermoepidermal junction. For C3d immunohistochemical staining, sensitivity, specificity, positive predictive value and negative predictive value were 86.66%, 100%, 100% and 80%, respectively, and for C4d immunohistochemical staining, respectively, were 73.3%, 100%, 100% and 66.66%. Conclusion: The immunohistochemical specificity of C4d and C3d on tissue blocks is the same as that of direct immunofluorescence test on fresh tissue, but it is less sensitive, so positive results for C3d and C4d immunohistochemical staining on paraffin blocks can be used to confirm the diagnosis of bullous pemphigoid.

In vivo evaluation of Sono-chemo therapy via hollow gold nanoshells conjugated to mitoxantrone on breast cancer.

Objectives: Conventional methods of cancer treatment include surgery, chemotherapy, radiation therapy, and immunotherapy. Chemotherapy, as one of the main methods of cancer treatment, due to the lack of targeted distribution of the drug in tumor tissues, is not able to destroy cancer cells and also affects healthy tissues and causes serious side effects in patients. Sonodynamic therapy (SDT) is a promising strategy for non-invasive treatment of deep solid cancer tumors. In this study, for the first time, the sono-sensitive activity of mitoxantrone was investigated and then mitoxantrone (MTX) was conjugated to hollow gold nanostructure (HGN) to improve the efficiency of in vivo SDT. Materials and Methods: Firstly, after the synthesis of hollow gold nanoshells and the PEGylation process, conjugation of MTX was performed. Then, after evaluating the toxicity of the treatment groups in vitro, in order to perform an in vivo study, 56 male Balb/c mice that had been tumorized by subcutaneous injection of 4T1 cells were divided into eight groups of breast tumor model. Ultrasonic irradiation (US) conditions including intensity of 1.5 W/cm2 (with a frequency of 800 kHz, 5 min), MTX concentration of 2 µM, and HGN dose of 2.5 mg/kg (unit of animal weight) were used. Results: The results show that administration of PEG-HGN-MTX caused a slight reduction in tumor size and growth compared with free MTX. Ultrasound also improved the therapeutic effect of the gold nanoshell in treated groups, and the HGN-PEG-MTX-US treated groups were able to significantly reduce and control tumor size and growth. Conclusion: The findings also show that MTX and HGN can be used as sonosensitizers in SDT. Also, HGN-PEG-MTX can act as a sono-chemotherapy agent for the combination of sonodynamic therapy and chemotherapy for in vivo breast tumors.

Potential protective effects of chrysin against immunotoxicity induced by diazinon.

Acute intoxication with diazinon (DZN) as a pesticide causes mortality and morbidity annually. This study shows the impact of sub-acute toxicity of DZN 20 mg/kg and the protective activities of chrysin (CH) as a flavone under the flavonoids family (12.5, 25 and 50 mg/kg) were assessed on BALB/c mouse immune system. The changes in morphological and functional properties of the immune system on thymus, spleen and liver histopathology, sub-populations of T lymphocytes, cytokines levels, transcription factors, complement function, phagocytosis, specific and total antibody productions were considered. The histopathological effects of DZN on the spleen and thymus were not significant, but the liver was damaged remarkably. In the presence of CH, the toxic effect of DZN is suppressed. DZN significantly decreased the number of whole blood TCD4+, TCD8+ and NK cells and suppressed the phagocytosis, delayed-type hypersensitivity (DTH) responses to sheep red blood cell (SRBC). Furthermore, it suppressed specific anti-SRBC-Ab, total IgG and IgM production, T-bet expression, and IFN-γ production. In contrast, DZN did not significantly affect complement function and the number of NK cells, TCD4+ and TCD8+ splenocytes. However, it potentiated the expression of GATA-3, ROR-γt and FOXP3 gene expression and consequently produced IL-4, IL-10, IL-17 and TGF-ß in whole blood. CH not only significantly increased the variables mentioned above at 12.5, 25 and 50 mg/kg but also could overcome the toxic effects of DZN on whole blood lymphocyte sub-populations and specific and total Ab production in 25 and 50 mg/kg concentrations, phagocytosis and DTH responses in 50 mg/kg, and modulation of the transcription factors and cytokine production, mainly in 25 and 50 mg/kg. In conclusion, DZN in sub-acute doses could remarkably deteriorate immune responses. However, CH can overcome the toxic effects of DZN on the immune components and functions of the immune system.

Non-collagenous extracellular matrix protein dermatopontin may play a role as another component of transforming growth factor-ß signaling pathway in colon carcinogenesis.

OBJECTIVES: Dermatopontin (DPT) is an extracellular matrix protein that plays roles in increasing the activity of transforming growth factor-ß (TGF-ß) and induction of cell quiescence. These roles suggest a tumor suppressor function for DPT. This study aimed to investigate changes in DPT gene expression in colorectal cancer providing a better understanding of its carcinogenesis. MATERIALS AND METHODS: We used Matched Tumor/Normal Expression Array and Cancer Profiling Arrays I containing 34 and 7 cases of colorectal cancer and their matched controls, respectively, to test DPT expression. In addition, 38 newly diagnosed cases of colorectal cancer were enrolled and their fresh colonic tumoral and normal specimens were obtained. DPT mRNA expression was analyzed using real-time PCR. In cases with DPT under expression, exonic regions of the DPT gene were sequenced using the Sanger method. RESULTS: In array samples, DPT expression was decreased in 82.9% (34/41), increased in 12.2% (5/41), and had no changes in 4.9% (2/41). DPT was decreased in 14 fresh samples (36.8%), while 12 cases (31.6%) showed overexpression and the others had no changes. DPT expression showed no significant difference among various tumor grades and stages. The frequencies of DPT overexpression were higher in tumors having lymph node involvement (47.7% vs 28%, P=0.59). In 2 cases mutations were detected that may be responsible for decreased expression of DPT. CONCLUSION: The similarities between changing patterns of DPT and TGF-ß expression in colorectal cancer demonstrate that DPT may act as a pre-receptor component of the TGF-ß signaling pathway in colon carcinogenesis.

The Prognostic Role of Tumor Marker CA-125 in B-Cell non-Hodgkin's Lymphoma.

BACKGROUND: B-cell non-Hodgkin's lymphoma (NHL) is a common malignancy of lymphoid tissues. Different types of NHL show various behaviors, prognoses, and responses to treatment. Evaluation of disease activity in NHL can be helpful in managing and even increasing the patient's survey. METHODS AND RESULTS: In total, 121 patients (76 males and 45 females), and their age range were 18-53 years, were evaluated in this study. The mean level of serum carbohydrate antigen 125 (CA-125) was 89.3±18.5 u/ml, ranging from 27 to 135 u/ml. There were significant differences in International Prognostic Index (IPI) score (p=0.002), stage of the disease (p=0.006), mortality rate (p=0.02), and relapse rate (p=0.04) between patients with serum CA-125 level <35 u/ml and patients with CA-125 level >35 u/ml. CONCLUSION: CA-125 seems to be a useful and reliable tumor marker for monitoring a patient with NHL. It might be the time to consider CA-125 in staging, prognostic scoring, or decision making about NHL treatment.