Journal of Cytology & Histology

Aim: Bone marrow stromal cells (BMSCs) are a promising candidate for cell therapy in ischemic stroke. However, the majority of BMSCs are readily lost after transplantation because of apoptosis due to ischemia-reperfusion (I/R) injury. In the present study, we aimed to evaluate the effects of hypoxia preconditioning (HP) on simulated I/R injury in cultured BMSCs.
Methods: Four generations of BMSCs were used as subjects. BMSCs were divided into 6 groups, including normal group, I/R group(I/R followed 0 h HP), 2 h-HP group(I/R followed 2 h HP ), 4 h-HP group(I/R followed 4 h HP), 6 h-HP group (I/R followed 6 h HP) and 8 h-HP group(I/R followed 8 h HP) 2 h-HP group(I/R followed 2 h HP ), 4 h-HP group(I/R followed 4 h HP), 6 h-HP group (I/R followed 6 h HP) and 8 h-HP group(I/R followed 8 h HP). BMSCs were subjected to HP by exposing the cells to hypoxia (2%O2). After 12 h of reoxygenation, BMSCs were suffered from 3 h ischemia (<0.5% O2 and serum deprivation) and subsequent reperfusion (I/R). Cell viability, hypoxia-inducible factor (HIF) 1-α, caspase-3 and apoptosis were tested using MTT, ELISA and immunofluorescence staining respectively.
Results: There were no changes in cell viability after 2 h to 8 h HP in cultured BMSCs as compared to the normal group. But HP upregulated the content of HIF-1α in BMSCs and protected the BMSCs from later I/R injury, as evidenced by increased cell viability and decreased Caspase-3 release and apoptosis, especially in 8 h-HP group.
Conclusion: HP attenuated the apoptosis induced by I/R injury via co-regulating the expression of HIF-1α and Caspase-3 in BMSCs, and thereby might play a beneficial effect on cell therapy

Background and objective: 24 patients, all of them vaccinated in postpubertal period, on their own descision, all of them had abnormal PAP tests. In all of them an expert colposcopy was performed, in all of them a low and high grade cervical lesion was detected. HPV genotypization, punch biopsy of the lesion folowed by histology and immunohistochemical staining of p16 INK4a were performed.
Study design: Our study concerns 24 patients suffering of cervical lessions. The age of these patients was 20- 34 years. 20 of them were vaccinated by Silgard and 4 of them by Cervarix.
Results: Two patients were hrHPV negative, in 21 patients the cervical lesions were infected with different HPV combinations, only in one woman single hrHPV type 52 was present. The frequent hrHPV types occurence was: 52/11x/, 39/11x/, 31/9x/, 45/6x/, 51/6x/.
Conclusions: Different high risk HPV combinations were present in 21 of 24 examined patients. The p16INK 4a positivity was detected in all lesions in all women examined.

The present study includes ultrastructural aspects of molluscicidal effects of wild botanic aqueous extract, namely Za’ater plant (Origanum syriacum) on the gametogenesis of two terrestrial slugs, giant grey slug Limax maximus and tree slug L. marginatus. L. marginatus was more susceptible to this plant than L. maximus (LC90 are 445.1 and 460.2 for L. marginatus and L. maximus respectively). The present results revealed that Za’ater’s extract had adverse effects on gametogenesis of the treated slugs, including marked inhibition of the various stages of gametogenesis, besides intracellular accumulation of the toxic agents of Za’ater-plant. Therefore, the treatment of the tested slugs with LC90 of Za’ater extract can be recommended for controlling such slugs in the field, instead of using chemical pesticides that continuously pollute the environment.

We describe a 64-year-old caucasian man with squamous cell carcinoma of the base of the tongue that metastasized to the choroid. The primary tumor was known to be high risk HPV positive and initially diagnosed as T3N3M0 stage IVB two years prior to presentation. Funduscopy revealed an 18 x 17 x 8.9 millimeter mushroom shaped amelanotic tumor with intrinsic vascularity. Ultrasonography revealed medium to high internal reflectivity with echolucent pockets and a positive angle kappa. Marked intrinsic vascularity and associated retinal detachment was also observed. Fine needle aspiration biopsy was performed and 40cGy where delivered via plaque brachytherapy. Histopathologic and cytologic findings of the primary tumor and of the choroidal tumor, respectively are discussed in detail.

Background: Artificial neural network (ANN) has been used in medicine to predict either the treatment or the investigative outcomes. The aim of this study was to validate the use of ANN models for predicting recurrence in non muscle invasive bladder cancer (NMIBC) treated by Bacillus Calmette Guerin (BCG) immunotherapy.
Materials and Methods: In this study, we developed a Multilayer Percepteron (MLP) based ANN to detect recurrence in NMIBC through the analysis of histopathologic data. The study includes 308 patients (mean age, 63.92 years; range, 31-92 years) who were treated with transurethral resection followed by BCG-immunotherapy. Time follow-up was 30 months.
Results: In the test group, 39 out of 40 cases were correctly classified by the MLP base neural network with an optimum Mse error (0.02634). Only one case was classified as false positive, with no false negative results. The sensitivity, specificity, positive predictive and negative predictive values calculated from the output data were 96.66%, 100%, 100% and 90.9% respectively. Network can predict the outcome of 79% (34*100/35) of patients in the testing data set correctly.
Conclusion: The proposed algorithm produced high sensitivity and specificity in predicting the recurrence in NMIBC after BCG immunotherapy compared to conventional statistical analysis. Therefore the use of ANNs will increasingly become the method of choice to calibrate complex medical models.

Methotrexate (MTX) had been used for many years and complications usually encountered during treatment especially in cancer patients. The aim of the present study was to determine the preventive and early-stage anti cytotoxic effects of green tea on the histology of the rats cheek mucosa treated by high single dose of methotrexate. The study included 36 Albino rats. Twelve animals were used in the pilot study to find the maximum toxic dose, and the other twenty four were divided into four groups: vehicle treated control group, green tea extract treated control group (40 mg/kg/day), Methotrexate treated group (80 mg/kg), and methotrexate and green tea extract treated group. Cheek mucosa excision was then performed. Histopathological examination was performed with hematoxylin-eosin, PAS and Masson’s trichrome stains. Cell proliferation was examined using the Ki-67 antibody and anti apoptotic effect was determined based on Bcl-2 staining. The result showed that in the methotrexate and green tea extract treated group, a significant increase in the thickness of the epithelium (p<0.01), a significant decrease in the number of congested blood vessels in the connective tissue of rat cheek mucosa (p<0.01), a non significant different in the Ki-67 expression (p>0.01), and a significant increase in Bcl- 2 expression (p<0.01) were seen in comparison with the methotrexate treated group. In conclusion green tea extract at a concentration of (40 mg/kg/day) produced protection against methotrexate induced oral mucositis by its antioxidant, antiangiogenic and anti apoptotic activity effects, and therefore can be used as a protective natural product to oral mucosa against methotrexate induced cytotoxicity.

Toll-like receptors (TLR) signaling plays an important role in the B cell biology. It induces the maturation, proliferation and antibody production in response to pathogen recognition. Interleukin-1 Receptor-associated Kinase-4 (IRAK-4) is TLR downstream molecule which is of strategic importance in starting a cascade of NF-kappaB (Nuclear factor-kappaB) and MAPK (Mitogen-activated protein kinase) intracellular signaling pathways. The aim of the study was to identify the differences in IRAK-4 expression in normal and chronic lymphocytic leukemia (CLL) lymphocytes. We have also studied the IRAK-4 expression alteration upon TLR2/4 ligand stimulation in leukemic and normal B cells. We carried out Real-Time PCR and Western blot analysis of IRAK4 mRNA (RQ) and protein expression. The study were performed on the isolated CD19+ normal B lymphocytes and CLL cells (CD5+CD19+) before and 30 min after (Lipopolysaccharide) LPS and (Staphylococcus aureus strain Cowan I) SAC stimulation. IRAK-4 mRNA expression was significantly higher in normal CD19+ lymphocytes as compared to leukemic CD5+CD19+ subpopulation (RQ median 0.90 vs 0.54; p=0.012). In normal lymphocytes, LPS and SAC treatment led to significant IRAK-4 down-regulation both at mRNA (RQ median 0.90 vs 0.53 vs 0.40; p=0.03, p=0.01) and protein level, whereas generally there were no significant changes in IRAK-4 expression neither on mRNA nor protein level. No trend or significant differences in IRAK-4 expression in leukemic lymphocytes after LPS and SAC stimulation may imply the presence of alleged defect of immune tolerance in CLL.

Introduction: Differentiation between adenocarcinoma of the lung (LADC) from the other non-small cell lung carcinoma (NSCLC) is possible by immunohistochemistry. When diagnosis of LADC is established, the rest of lung cancer tissue could be used for molecular testings, particularly EGFR analysis
Aim: To evaluate 4 antibodies TTF-1, Napsin-A, Cytokeratin7 and Surfactant B in diagnosis of LADC.
Materialsand Methods: Fifty small-sized samples, obtained upon bronchoscopy or transthoracic fine-needle lung biopsy, were included in this study. Diagnosis of LADC was performed after routine hematoxylin-eosin examination and TTF-1, Napsin-A, Surfactant B and Cytokeratin7 immunohistochemistry. Descriptive statistical method (%) was used in the study.
Results: Cytokeratin7 was expressed in 90% (45/50), than TTF-1-86% (43/50) and Napsin-A - 82% (41/50) and Surfactant B in 6% (28/50) in LADCs. Conclusion: TTF-1 is the most useful for diagnosis of LADC and Napsin-A in cases where TTF-1 is not expressed. Cytokeratin7 is useful in differentiation from digestive adenocarcinoma and Surfactant B in cases where TTF-1 and Napsin-A, individually or both are not expressed.

Background: The heat shock protein 90 (HSP90) is overexpressed and highly associated with poor prognosis in many malignancies. However, the role of HSP90 in Non muscle invasive bladder cancer (NMIBC) has not been thoroughly elucidated. The aim of this study is to investigate the relationship of HSP90 expression with the response to BCG immunotherapy of NMIBC.
Methods: HSP90 expression was evaluated by immunohistochemistry in 43NMIBC samples. The relationships between HSP90 and the response to BCG immunotherapy were analyzed. The results were correlated with clinicopathological characteristics using univariate and multivariate analyses.
Results: Univariate Cox regression analysis of baseline characteristics and HSP90 expression showed that no significant association was found with BCG immunotherapy response (p=0.894). In the other hand, multivariate Cox regression analysis showed that HSP90 protein is not an independent factor of tumor response after BCG immunotherapy (p=0.884).
Conclusion: This study demonstrates that HSP90 could not be a useful prognostic marker for BCG treatment in NMIBC. We emphasize that this was a preliminary study and therefore further confirmation on a larger set of tissues is necessary.

Gastrointestinal involvement in disseminated histoplasmosis can occur at any site along the GI tract without specific manifestations, although causes clinical symptoms in only 3–12% of patients. Intestinal histoplasmosis is the most common one, particularly at the terminal ileum as it’s abundant in lymphoid tissue. Endoscopic findings in the intestines were from erythema, edema, ulcers to perforation. As the lack of specificity of these manifestations in symptoms and colonoscopy, intestinal histoplasmosis may be misdiagnosed, leading to inappropriate therapies and unnecessary surgical interventions. We report a case of immunocompetent disseminated histoplasmosis involving the colon and terminal ileum which presented fever and weight loss without gastrointestinal symptoms. So taking disseminated histoplasmosis as differential diagnosis even in immunocompetent asymptomatic patients is necessary for gastroenterologists and endoscopists.

The present work investigated histological and cytological studies, using SEM, included sensory organs of flatworm Bipalium kewense. The shovel-headplate of B. kewense is limited all around its margin by distinct sensory organs, consisting of a row of sensory papillae, pits and eyespots. Sensory papillae and sensory pits are situated at the peripheral edge of worm-head whereas numerous eyespots are located within the margin of the dorsal shovelheadplate and along the body margin. Each eye is composed of pigment cells surrounding half moon-like lens and retinal cells. The present results suggested that the sensory papillae and pits are chemo-receptors.

Myeloid sarcoma is a rare malignant disease defined as extramedullary infiltration of immature myeloid cells and may precede or occur synchronously with acute myeloid leukemia. We reported a 35-year-old male of isolated myeloid sarcoma who presented with symptoms similar to chronic rhinosinusitis initially. Isolated myeloid sarcoma in the paranasal sinuses is exceedingly rare after reviewing the previous literatures. Through proper histological, adequate panels of immunohistochemical stain (positive markers including CD68/KP1, myeloperoxidase, CD 117, CD 99, CD68/PG-M1, lysozyme, CD34, terminal deoxynucleotidyl transferase and negative markers including CD3, CD20, CD45RO and CD79a) and cytogenic study lead to accurate diagnosis. Early intervention with systemic chemotherapy with cytarabine-based regimens is the treatment of choice. The role of chromosomal aberrations and genetic abnormality related to prognosis remain uncertain.

Background: 2 - hydroxyethyl methacrylate (HEMA) is one of the common components of most resin-based dental materials. Various studies have shown that HEMA can diffuse through dentin due to its low molecular weight and its hydrophilicity, and can affect the underlying odontoblast, cell division and activity. In this work, we have studied the influence of HEMA in regulating the expression of pro-collagen α1 type I and tenascin - C proteins in human fibroblasts after long term and low concentrations of HEMA.
Methods: Human dental pulp cells were exposed to 0.1 mM and 0.5 mM for 1, 3, 5, 7, and 15 days. MTT assay, immunofluorescence and western blot analysis were carried out to investigate cell viability and modification in collagen type I and tenascin – C protein expression.
Results: MTT assay showed an high cell viability, western blot and immunofluorescence demonstrated a down-regulation of collagen type I protein and un up-regulation of tenascin - C protein, the latter involved in cellular stress.
Conclusion: low concentrations and long-term HEMA exposition, greatly influences the expression of collagen type I protein and tenascin – C protein in human dental pulp cells, modifying the extracellular matrix toward a stressful microenvironment.

Introduction: Large mastoid cholesterol granulomas, particularly aggressive ones, are rare. We present the first case of a mastoid cholesterol granuloma with a nonspecific clinical presentation, and aggressive behavior, destroying the petrous internal cortical plate, dura mater, and sigmoid sinus wall. Radiographic and histological examinations can help to distinguish such granulomas from others.
Case presentation: A 50-year-old male presented with left otalgia, left blood-stained otorrhea, and a left temporal headache radiating to the mastoid. The radiographic examination revealed a left-sided, soft-tissue mass extending into the posterior fossa and compressing the sigmoid sinus inward. The histopathological features were consistent with a cholesterol granuloma. He underwent a cortical mastoidectomy to explore the lesion and macroscopic complete resection was achieved.
Conclusion: This is the first case of a mastoid cholesterol granuloma eroding the dura and sigmoid sinus. This case illustrates the need to consider cholesterol granuloma in daily clinical practice. Such lesions can exhibit noncharacteristic features and have an aggressive nature.