ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 54 No. 4, 2013

1. Human adipose-derived stem cells: definition, isolation, tissue-engineering applications

S. Nae, I. Bordeianu, A. T. Stancioiu, N. Antohi

Recent researches have demonstrated that the most effective repair system of the body is represented by stem cells - unspecialized cells, capable of self-renewal through successive mitoses, which have also the ability to transform into different cell types through differentiation. The discovery of adult stem cells represented an important step in regenerative medicine because they no longer raises ethical or legal issues and are more accessible. Only in 2002, stem cells isolated from adipose tissue were described as multipotent stem cells. Adipose tissue stem cells benefits in tissue engineering and regenerative medicine are numerous. Development of adipose tissue engineering techniques offers a great potential in surpassing the existing limits faced by the classical approaches used in plastic and reconstructive surgery. Adipose tissue engineering clinical applications are wide and varied, including reconstructive, corrective and cosmetic procedures. Nowadays, adipose tissue engineering is a fast developing field, both in terms of fundamental researches and medical applications, addressing issues related to current clinical pathology or trauma management of soft tissue injuries in different body locations.

Corresponding author: Sorin Nae, MD; e-mail: naedaniel@gmail.com, naesorin@yahoo.com

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2. Lymphatic microvessels density, VEGF-C, and VEGFR-3 expression in 25 cases of breast invasive lobular carcinoma

Maria Ciobanu, Irina Anca Eremia, Stefania Craitoiu, Cl. Margaritescu, A. Stepan, V. Patrascu, C. C. Georgescu, Daniela Cernea, Daniela Dumitrescu

Invasive lobular carcinoma (ILC) is the second most common type of invasive breast cancer, having distinct morphologically but also prognostic and therapeutic features. This type of breast cancer shows a higher rate of multiple metastases with a more frequent axillary-lymph-node involvement. Related to these dissemination and metastatic features, we aimed to study the immunohistochemical expression of D2-40, VEGF-C and VEGFR-3 in 25 cases of ILCs stratified according to the histopathological and molecular classification. Regardless of histopathological or molecular subtype, the statistical tests proved that for ILC, the highest D2-40 lymphatic microvessels density (LMVD) was in the peritumoral areas. In classical subtype, the LMVD values were positively correlated with the degree of tumor differentiation and pTNM clinical stages and when these cases were classified based on the molecular criteria the highest recorded values were found in the luminal B subtype. In addition, regardless of the histopathological and molecular subtypes, the D2-40 LMVD varied in the same direction for both VEGF-C and VEGFR-3 categories, with the highest LMVD values recorded in those cases with the highest VEGF-C and VEGFR-3 reactivity, especially in the peritumoral areas. Considering only the molecular luminal A and B subtypes, we have noted that VEGF-C and VEGFR-3 expression was significantly higher in luminal A subtype compared to luminal B. This immunoprofile suggests the existence of a tumor type-specific lymphangiogenesis that may have certain prognostic and therapeutic implications.

Corresponding author: Claudiu Margaritescu, Associate Professor, MD, PhD; e-mail: c_margaritescu2000@yahoo.com

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3. The assessment between IL-6 and IL-8 and anthropometric status in malnourished children

Cristina Oana Marginean, Lidia Man, Ana Maria Pitea, A. Man, Cl. Marginean, O. S. Cotoi

Purpose: To evaluate the correlations between the pro-inflammatory interleukins IL-6 and IL-8 and the anthropometric measurements in malnourished vs. non-malnourished children. Patients and Methods: We have examined 219 children from Pediatric Clinic I, University of Medicine and Pharmacy of Tirgu Mures, Romania, during January 1, 2012-March 1, 2013 and divided according to Body Mass Index (BMI kg/m2) in the following two groups: 164 with normal nutritional status - control group (BMI between -2SD and +2SD), and 55 children with malnutrition (BMI <-2SD). All the children were evaluated anthropometric: BMI, weight for age (W/A), height for age (H/A), mid-upper-arm circumference (MUAC), tricipital skinfold (TSF) and paraclinical: IL-6 and IL-8 levels. Results: From 219 children, 25.1% were malnourished. The mean age was 5.16 years in malnourished. IL-6 and IL-8 mean levels were 2.54 pg/mL, respectively 6.83 pg/mL in malnourished and 6.02 pg/mL, respectively 9.06 pg/mL in non-malnourished. By statistically comparing IL-6 in malnourished group vs. control group, we observed decreased values (p<0.0001) and also significantly lower values for IL-8. We also obtained statistical differences between the two groups in BMI, W/A, MUAC and TSF. The BMI SD have an increasing trend line, from -4SD in newborn malnourished to -2SD in near 18-year-old malnourished; the trend line had only a slight ascension in non-malnourished children. Conclusions: The interleukin levels and BMI, W/A, MUAC and TSF are significantly lower in malnourished children than in non-malnourished. This functional impairment may be involved in the malnutrition to develop a specific immune response in these children.

Corresponding author: Adrian Man, Lecturer, MD, PhD; e-mail: adrian.man@umftgm.ro

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4. Expression of CXCR4, MMP-13 and beta-catenin in different histological subtypes of facial basal cell carcinoma

M. E. Ciurea, Daniela Cernea, C. C. Georgescu, O. S. Cotoi, V. Patrascu, H. Parvanescu, D. Popa, V. Parvanescu, Raluca Niculina Ciurea, R. Mercut

Basal cell carcinoma (BCC) is one of the most common skin neoplasms in humans, accounting for almost 80% of all non-melanoma skin cancers worldwide. The nodular and infiltrative-morpheaform are the most common BCC types in the head and neck region and together with the micronodular subtypes are the most aggressive tumors, because of their tendency to infiltrate the deep subcutis, muscles and even bones. To explain the local aggressive behavior and their metastatic potential, many studies have been performed to identify the molecular determinants implicated in BCC tumor progression. For this reason, we investigated the immunohistochemical expression of CXCR4, MMP-13 and beta-catenin expression in six metatypical, eight infiltrative-morpheaform, six micronodular and five superficial facial BCCs. For all three markers, the tumor reactivity varied with the histological type. The highest reactivity was observed in metatypical subtype, especially at the level of areas with squamous cells differentiation. The lowest reactivity was recorded in micronodular and superficial BCC subtypes. Regardless histological subtype, the tumor reactivity was higher at the advancing edge and additional a strong stromal reaction was noticed for all investigated markers peculiar in fibroblasts, inflammatory cells and endothelial cells. All these data proved the utility of CXCR4, MMP-13 and beta-catenin immunohistochemical investigation in BCCs both for identification of high-aggressive tumors and to develop novel more efficient therapeutic strategy for these patients by targeting these biomarkers.

Corresponding author: Ovidiu Simion Cotoi, Lecturer, MD, PhD; e-mail: ovidiu.cotoi@umftgm.ro

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5. Electrical conduction system apoptosis in type II diabetes mellitus

S. Hostiuc, Alina Popescu, Elena Daniela Gutu, M. C. Rusu, F. Pop

Even though apoptosis is known to be associated with various cardiovascular pathologies, its presence in cardiac nodal tissue in adults was only scarcely researched. Cardiomyocyte apoptosis was associated with diabetic cardiovascular pathology. Our main objective was to test whether programmed cell death is present in nodal tissue in type II diabetes mellitus and, if present to characterize it. The study was designed as a qualitative one. We used autopsy samples of hearts from 10 patients (56 to 73-year-old, 6:4 male to female ratio), positive for type II diabetes mellitus. Samples from sinoatrial and atrioventricular nodes were stained with Hematoxylin-Eosin. For immunohistochemistry, we used primary antibodies for caspases 3 and 9, cathepsin B, and TRADD. Nodal tissue in all samples was characterized by diffuse interstitial fibrosis and chronic ischemic lesions; nuclear damage and foci of irreversible ischemic necrosis intermingled with isles of relatively morphologically normal myocytes. Sinoatrial and atrioventricular nodes were caspase-3 and -9 positive, and also cathepsin-B-positive, suggesting an overlap between apoptotic and necrotic mechanisms. Central area of the sinus node seemed to have the most severe lesions. As a conclusion, nodal apoptosis is present in nodal tissue in type II diabetes mellitus; it involves the intrinsic pathway and associated concomitant and/or post-apoptotic necrosis.

Corresponding author: Mugurel Constantin Rusu, Associate Professor, MD, PhD, Dr.Hab.; e-mail: anatomon@gmail.com

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6. Estrogen and progesterone receptor expression in the mammary gland tumors

M. Moise, M. M. Buruian, C. Ilie, Carmen Lacramioara Zamfir, Roxana Folescu, A. G. M. Motoc

In the primary mammary malignant tumors, including in situ carcinoma, it is recommended the carrying out of immunohistochemical diagnosis for the estrogen (ER) and the progesterone receptors (PR). We have studied the ER and the PR expression in malignant tumors, trying to identify the corresponding phenotypes according to the presence of these tumors. We have carried out a study on a total number of 80 carcinomas, divided into two groups: the first one constituted of 54 cases of carcinomas on which we had clinical data, and another group, constituted of 26 cases of mammary carcinoma, where no clinical data was available. We have observed that the values and the distribution of the ER and PR taken from the biopsies made in the patients with mammary carcinoma are influenced by the age and menopausal status. The combination of the ER/PR results lead to the definition of many tumoral phenotypes.

Corresponding author: Roxana Folescu, University Assistant, MD, PhD; e-mail: roxanafolescu@yahoo.com

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7. The involvement of growth differentiation factor 5 (GDF5) and aggrecan in the epithelial-mesenchymal transition of salivary gland pleomorphic adenoma

Anca Stefania Enescu, Cl. Margaritescu, Monica Mihaela Craitoiu, Aurelia Enescu, Stefania Craitoiu

Pleomorphic adenoma is the most common salivary gland tumor with annual incidence of 2-3.5/100 000 in population. The histogenesis of salivary gland pleomorphic adenoma is still unclear. One concept sustains the existence of an epithelial-mesenchymal transitions (EMT) process in pleomorphic adenomas by which neoplastic epithelial cells transdifferentiate into mesenchymal cells and leading to tissue heterogeneity from this salivary gland neoplasia. Here we investigate by immunohistochemistry the expression of growth differentiation factor 5 (GDF5) and aggrecan in 15 cases of salivary gland pleomorphic adenomas. We found that both markers were present in normal salivary gland, mainly in the cells that line striated and intercalated ducts suggesting their involvement in the morphogenesis of this duct system. A constant positive reactivity for both markers was recorded in transition regions from tubular proliferative units to myxoid areas suggesting the involvement of an EMT process during the tumorigenesis of such salivary gland neoplasia. Also, both markers may be implicated in the transdifferentiation of neoplastic myoepithelial cells from the myxoid zones to lacuna cells of adjacent chondroid areas completing the morphology of this salivary gland tumor.

Corresponding author: Claudiu Margaritescu, Associate Professor, MD, PhD; e-mail: c_margaritescu2000@yahoo.com

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8. Morphometric and ultrasonographic study of the human fetal hip joint during intrauterine development

Beata Baroti, Zsuzsanna Pap, Z. Panti, M. M. Buruian, Z. Pavai

The main method for the early screening of the developmental dysplasia of the hip (DDH) is the ultrasound imaging. There are several studies about the ultrasound imaging of newborns' hips, but only a few studies include the prenatal period of life. Our aim was to examine the prenatal development of the hip joint through the evolution of the alpha angle seen on the ultrasound, described in the Graf R method, combined with anatomical dissection. Materials and Methods: Thirty-one post-mortem fetal hips were analyzed trough anatomical dissection, in 25 cases trough ultrasound imaging, in which the alpha angle was measured. Based on the morphometric examination, we applied the sine rule and we calculated the alpha1 angle, which also represents the coverage of the femoral head. Results: Based on the morphometric examination, not only the diameters of the femoral head and of the acetabulum, but also the joint cavity (X) showed an increase during development. Both of the alpha angles (measured alpha, calculated alpha1) showed a decrease as the fetus developed. Conclusions: The decrease of the angles (alpha, alpha1) and the increase of the joint cavity during development correspond to the findings of the main research papers: the hip joint is less stable in the perinatal life. The alpha angle can be accurately determined only after the ossification of the acetabulum had started, in our case after the fetus is older than 18 weeks.

Corresponding author: Zsombor Panti, Student, General Medicine 6th year; e-mail: panti.zsombor@gmail.com

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9. Histopathological and immunohistochemical study of hepatic stellate cells in patients with viral C chronic liver disease

A. G. Ionescu, Letitia Adela Maria Streba, C. C. Vere, M. E. Ciurea, C. T. Streba, Mihaela Ionescu, Maria Comanescu, Eugenia Irimia, Otilia Rogoveanu

Objective: Our aim was to establish a correlation between hepatic stellate cells (HSCs) activity within different areas of hepatic tissue and the degree of liver fibrosis, necroinflammation, and steatosis in patients with viral C chronic liver disease. Patients and Methods: We prospectively included 41 liver biopsies from patients with chronic hepatitis C or liver cirrhosis, prior to antiviral treatment. Our control group consisted of seven tissue samples, obtained from 10 to 38 weeks old embryos. We assessed the alpha-smooth muscle actin (alpha-SMA) expression using immunohistochemistry and a semi-quantitative scoring method. Results: We found a strong correlation (p<0.0001) between the number of activated HSCs and the stage of fibrosis in all examined areas. Necroinflammatory activity was significantly correlated (p<0.05) with the number of activated HSCs in perivenular area (p=0.014) and intermediate area (p=0.018), and strongly correlated (p<0.0001) in periportal and portal tracts and fibrous septa areas. We found no correlation between the degree of steatosis and the number of activated HSCs in the perivenular area (p=0.25), intermediate area (p=0.166) and in the periportal area (p=0.154); however, in the portal tracts and fibrous septa area we observed a significant correlation (p=0.022). Conclusions: The analysis of HSCs activity within specified areas of liver tissue may lead to new perspectives in early diagnosis of liver fibrosis and in the development of future antifibrinogenic therapies.

Corresponding author: Cristin Constantin Vere, Associate Professor, MD, PhD; e-mail: cc.vere.umf@gmail.com

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10. Valproic acid substantially downregulated genes folr1, IGF2R, RGS2, COL6A3, EDNRB, KLF6, and pax-3, N-acetylcysteine alleviated most of the induced gene alterations in chicken embryo model

Chiu-Lan Hsieh, Kuan-Chou Chen, Chi-Yang Ding, Wan-Jane Tsai, Jia-Fong Wu, Chiung-Chi Peng

Valproic acid induced teratogenicity at genetic and somatic levels, the action mechanism is still unclear. We hypothesized that folate receptor gene (folr1) and others may be interacting to elicit neural tube defect (NTD), while N-acetylcysteine (NAC) may be beneficial for protection. In chicken embryo model, the experiment was conducted in two parts. The first part was carried out to test the optimum dose of VPA. The second part was conducted to test the protective effect of NAC at doses 10 and 20 mM. VPA induced dysvascularization, incomplete somite enclosure, histone deacetylase (HDAC) inhibition, folate deficiency, homocysteine accumulation, SOD inhibition, glutathione depletion, elevated MDA and hydrogen peroxide. NAC alleviated most of these adverse effects. The microarray analysis revealed 17 genes downregulated and four upregulated. The relevancy covered translation (23%), signal transduction (23%), transcription (16%), cell adhesion (16%), neural cell migration (8%), transport (7%), and organismal development (7%). The genes insulin-like growth factor 2 receptor gene (IGF2R), regulator of G-protein signaling 4 gene (RGS4), alpha 3 (VI) collagen gene (COL6A3), endothelin receptor type b gene (EDNRB), and Krueppel-like factor 6 gene (KLF6) substantially downregulated in reality were directly intermodulating and associated with NTD. VPA downregulated folr1 gene in a dose responsive manner without affecting pax-3 gene, which was ascribed to the metahypoxic state. Conclusively, VPA affects 21 genes: 17 downregulated and four upregulated. VPA dose responsively downregulates gene folr1 without affecting pax-3 gene. These adverse effects can be partially alleviated by N-acetylcysteine.

Corresponding author: Chiung-Chi Peng; e-mail: misspeng@ms2.hinet.net

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11. A profile of lung carcinomas: study on 364 cases

Camelia Demetrian, A. Demetrian, J.-P. Meunier, N. Naffaa, M. Olaru, R. M. Plesea, I. E. Plesea

The authors have proposed to analyze retrospectively the clinical-morphological profile of a consistent group of lung carcinomas treated surgically. The studied material consisted of clinical and pathological medical records from 364 patients confirmed histopathologically with lung carcinoma after surgical intervention. Five main groups were defined based on the histopathological criteria and then compared. The assessment of clinical data, in spite of a wide range of clinical expressions, revealed some particular features for each of the defined groups. The morphological data outlined also different behavioral profiles for each of the histopathological types of lung carcinoma. These results showed that malignant epithelial tumors of the lung are still a major challenge from the detection until the therapeutic intervention and, therefore, the preoperative clinical-morphological investigation is crucial for a better adjustment of the therapeutic act according to the individual profile of each type of tumor.

Corresponding author: Iancu Emil Plesea, Professor, MD, PhD; e-mail: pie1956@yahoo.com

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12. Testicular histopathological changes following sodium fluoride administration in mice

Nicoleta Dimcevici Poesina, C. Balalau, Maria Barca, I. Ion, Daniela Baconi, C. Baston, Violeta Baran Poesina

It has been revealed that excessive fluoride intake on long-term is associated with toxic effects and can damage a variety of organs and tissues in the human body, including the male reproductive system. However, the molecular mechanisms of fluoride-induced male reproductive toxicity are not well understood. The study wants to get news concerning the effects of natrium fluoride on testicular tissues when this substance is administrated to a population of mice. The study was conducted on NMRI mice descending from the pregnant females treated with 0.25 mg and 0.5 mg natrium fluoride by daily gavage, from the beginning of pregnancy until the lactation is ceased. Then, the mice, males and females, were divided in six groups, three groups descending from the pregnant females treated with 0.5 mg natrium fluoride (Groups A, B and C) and three groups from the pregnant females with 0.25 mg natrium fluoride (Groups D, E and F). From the moment when the lactation is finished until the adulthood, the animals received the following treatments: homeopathic (a CH7 solution of natrium fluoride - Groups A and D), allopathic-homeopathic (0.25 mg/1000 natrium fluoride administered like drinking water ad libitum and CH7 solution of natrium fluoride - Group E; 0.5 mg/1000 natrium fluoride administered like drinking water ad libitum and CH7 solution of natrium fluoride - Group B), and allopathic administration of natrium fluoride (0.25 mg/1000 natrium fluoride like drinking water ad libitum - Group F; 0.5 mg/1000 natrium fluoride like drinking water ad libitum - Group C). When the males reached the adulthood, the administration of natrium fluoride was stopped and, by randomization, they where selected for euthanasia. The euthanasia was realized by cervical dislocation. The testes for the histopathological examination were preserved in a 10% formalin solution. The preparation of samples for optical microscopy was realized with Hematoxylin-Eosin staining. The results indicate that natrium fluoride administered in different doses, even at homeopathic dose or at allopathic-homeopathic dose, determined vacuolar dystrophy of epididymal epithelial cells, vacuolar dystrophy of linear seminal cells and necrosis.

Corresponding author: Cristian Balalau, Senior Lecturer, MD, PhD; e-mail: dr.balalau@gmail.com

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13. Infrastructure of the telocytes from tumor stroma in the skin basal and squamous cell carcinomas

N. Mirancea, Ana-Maria Morosanu, G.-V. Mirancea, F. D. Juravle, V. S. Manoiu

In this paper, we focus our interest on the ultrastructure of telocytes (TCs) present inside of tumor-stroma in basal cell carcinoma (BCC) and squamous cell carcinoma (SCC). Tumor-stroma cooperation is necessary for tumor growth, invasive behavior and ectopic development of microtumors. There is a plethora of reports about the role of different stromal cell types in tumor evolution in the human body. In this line, almost nothing is known about the recently identified interstitial cell type called telocyte (TC). To our best knowledge, this is the first study to publish TCs in malignant tumors, namely BCC and SCC. Here, we described the infrastructural aspects of TCs as well as their relationships with other tumor stroma components. TC from the tumor stroma has cell body where the nucleus is located and exhibits two (rarely more) very long cell extensions of tens (over 60-100 micro-m) termed telopodes. A telopode appears as an alternation of very thin segments called podomers and dilated segments called podomes, which accommodate mitochondria, rough endoplasmic reticulum, cytoskeleton, caveolae, as well as coated vesicles. TCs establish homocellular junctions leading to a 3-D network inside of peritumoral stroma. TCs may play an important role in intercellular signaling via stromal synapses and shed microvesicle transfer. Comparative evaluation with normal dermal skin showed that telocytes from tumor stroma have a very restraint number of heterocellular junctions. The limitation of TCs heterocellular junctions suggests a possible involvement in induction of cell-cell communication alterations into the peritumoral stroma and, consequently, into the whole tumor mass.

Corresponding author: Nicolae Mirancea, PhD, Senior Scientist Grade I; e-mail: nick_mirancea@yahoo.com

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14. The synergy between lasers and adipose tissues surgery in cervicofacial rejuvenation: histopathological aspects

Andreea Cristina Nita, Dana Mihaela Jianu, I. P. Florescu, Maria Filipescu, O. Cobani, S. A. Jianu, D. A. Chirita, Adriana Bold

Background and Aims: Nowadays patients want a long-lasting youthful appearance but through a less invasive approach. Our unique approach, "AdipoLASER reJuvenation (ALJ)", involves a variety of less invasive techniques based on simultaneous laser resurfacing, and lipolysis together with adipose tissue graft and redistribution. Recently, we added to this platelet-rich plasma therapy. We conducted a study aiming to emphasize the histopathological changes occurred following these procedures. Patients and Methods: Between 2011-2012, we included 50 patients that were preparing for abdominoplasty (tummy tuck) in which we applied ALJ procedures, in two comparative zones of inferior abdomen. Histological examination varied from 10 days to four months, according to the time of the abdominoplasty. Results: There was a notable histological difference between stimulated and non-stimulated fat graft regarding adipose cells structure and number, neocollagenesis, and dermal matrix remodeling. Conclusions: The low level laser therapy effect (LLLT) of the fractional CO2 laser combined with the effect of the growth factors derived from activated platelets (PRP) prolonged the life and improved the take of the facial fat graft, increase collagen formation and lead to a better remodeling of the dermal matrix. This unique surgical combination of all four approaches in our ALJ with additional PRP offers a real less invasive but strongly visible - yet natural result - as an alternative to the classic facelift.

Corresponding author: Dana Mihaela Jianu, Associate Professor, MD, PhD; e-mail: djianu02@gmail.com

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15. Effect of intraperitoneal administration of sterile human cerebrospinal fluid in rats

V. T. Grigorean, Aurelia Mihaela Sandu, Ruxandra Diana Sinescu, M. L. Riza, I. E. Plesea, C. D. Lupascu, M. Popescu

Introduction: Distal ventriculoperitoneal shunt failure can be attributed to unabsorbed cerebrospinal fluid (CSF) from peritoneum. The objective of the experiment was to determine peritoneal reaction in rats after intraperitoneal administration of human CSF and evolution of local inflammatory response. Materials and Methods: Wistar rats were used divided into four groups: three groups in which intraperitoneal injection of 3 mL, 2 mL and 0.5 mL of sterile human CSF was done and a control group. After sacrificing the animals, at 24, 48 or 72 hours, macro- and microscopic examination of the peritoneal cavity and peritoneal fluid analysis were performed. The experiment was done in compliance with legislation regarding animal research. Results: Administration of high dose CSF (3 mL) led to death of all specimens. The dose of 0.5 mL of sterile CSF intraperitoneally administered was compatible with survival. Peritoneal response varied from necrotic-purulent reaction, with maximum intensity in group 1, and milder in group 2, to minimum inflammation in small foci and polymorphic cells in group 3. Inflammation only partially resolved in some specimens from group 3 after 72 hours, which incriminates the role of unabsorbed peritoneal CSF in pathogenesis of abdominal complications of ventriculoperitoneal shunts. Conclusions: Intraperitoneal administration of sterile human CSF caused inflammatory response of varying degrees and with multiple locations. High doses of CSF led to death of specimens. At 24 hours, the peritoneal response ranging from congestion to purulent reaction was acute, intense and diffuse but after 72 hours, the inflammatory response was mild, focal and limited.

Corresponding author: Aurelia Mihaela Sandu, MD; e-mail: aurasandu@gmail.com, aurassandu@yahoo.com

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16. Evaluation of the wound-healing effect of a novel Hypericum perforatum ointment in skin injury

Anca Irina Prisacaru, C. V. Andritoiu, Corina Andriescu, E. C. Havarneanu, M. Popa, A. G. M. Motoc, Anca Sava

The present experiment aims to formulate and characterize a new phytotherapy ointment based on a total extract of Hypericum perforatum included in a novel ointment base. In order to investigate the healing properties of the ointment, in vivo experimental wound models of linear incision, circular excision and thermal burn were performed on Wistar rats. Topical treatment was achieved daily, for 21 days. Clinical and macroscopic evaluation, determination of wound contraction rate, period of re-epithelialization, and histopathological examination were achieved, along with the determination of the particle diameter and particle size distribution of the ointment. The results demonstrate that the tested novel ointment has significant wound healing effect in skin injuries and reveals to be safe for use.

Corresponding author: Anca Sava, MD; e-mail: dr_anca_sava@yahoo.com

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17. The triage of low-grade cytological abnormalities by the immunocytological expression of cyclin-dependent kinase inhibitor p16INK4a versus Human Papillomavirus test: a real possibility to predict cervical intraepithelial neoplasia CIN2 or CIN2+

Ruxandra Stanculescu, Elvira Bratila, Vasilica Bausic, Teodora Vladescu

Objective: Assessing the hypothesis that p16INK4a immunocytochemistry (ICC) has better relevance than Human Papillomavirus (HPV) testing at detecting high-grade cervical intraepithelial neoplasia (HGCIN) upon histopathological diagnosis in women with abnormal cytologies such atypical squamous cells of undetermined significance (ASC-US) or low-grade squamous intraepithelial lesions (LSIL). Materials and Methods: A retrospective study of 63 selected cases (22 with ASC-US and 41 with LSIL) was performed at "St. Pantelimon" Clinical Hospital, Bucharest, Romania, using p16INK4a ICC and Linear Array HPV Genotyping Test. All cases have been followed-up by colposcopy and biopsies. The sensitivity and specificity of p16INK4a and HPV were analyzed by chi-squared test. Results: LSIL cytologies were more likely to be p16INK4a positive than those with ASC-US: OR=3.1, 95% CI (1.06-9.11). The processed data show that in women with LSIL the sensitivity of p16INK4a is 37.5% higher than that of high-risk(hr)-HPV (p=0.0050), whereas in ASC-US it is 44.5% higher (p=0.0577). In ASC-US, p16INK4a has a higher specificity (84.62%) than hr-HPV (53.85%); for LSIL cytologies, this difference is less steep: 58.82% for p16INK4a as compared to 47.06% for HPV. Conclusions: The p16INK4a is significantly more sensitive than hr-HPV in both low-grade abnormal cytologies and has higher specificity than HPV testing to detect HGCIN, mainly in women with ASC-US cytologies. Only women with ASC-US and LSIL cytologies who test positive for p16INK4a should be directed to colposcopy and/or biopsy. p16INK4a is a suitable immunocytochemical marker which increases the accuracy of diagnosis at women with low-grade cytologic abnormality.

Corresponding author: Teodora Vladescu, MD, PhD, e-mail: teodoravladescu@yahoo.com; Ruxandra Stanculescu, MD, PhD, e-mail: ruxandra_v_stanculescu@yahoo.com

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18. A combined imagistic and morphological approach of lung tumors: study on 64 cases

M. Olaru, D. Malaescu, A. Demetrian, L. Ioncica, B. Stanoiu, Daniela Dragnei

Lung cancer is currently one of the major health problems, being considered one of the most common causes of death by cancer worldwide. That is why establishing as early as possible a diagnosis in order to start an appropriate therapy still a challenge. The studied group consisted of 64 patients who were investigated following the next algorithm: chest X-ray, bronchoscopy, guided bronchial brushing and/or bronchoalveolar lavage followed by cytological examination and if possible, endobronchial biopsy followed by histopathological examination. Patients were usually men, aged over 60 years coming from an urban area, smokers and with symptoms evoking the presence of neoplasia. Tumors presented as large tumoral masses, placed centrally and with obvious local spread, protruding or pushing intraluminally with subsequent bronchial stenosis, usually complicated with ulceration and bleeding, which proved to be, when histopathological examination was possible, firstly squamous carcinomas and then small cell carcinomas. The imagistic investigation in conjunction with the cytological evaluation can establish the diagnosis of malignancy of centrally located lung tumors in almost 80% of cases. The diagnosis can be improved by increasing the number of biopsy sampling and/or by the immunohistochemical marking of the cytological samples.

Corresponding author: Marian Olaru, Associated Professor; e-mail: olarumarian1952@yahoo.com

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19. Clinical aggression of prolactinomas: correlations with invasion and recurrence

Mihaela N. Popescu, Elena Ionescu, L. C. Iovanescu, B. V. Cotoi, A. I. Popescu, Anca Elena Ganescu, Adina Glodeanu, Cristiana Geormaneanu, Andreiana Moraru, Anca Patrascu

Some hypophyseal adenomas are discrete, well-marked lesions, which can be curatively removed by surgical resection, while others seem aggressive, invasive, recidive-prone, most often progressing in spite of any pharmacological, surgical or radiotherapeutical interventions. There is also a great variability within the incidence of aggression and invasion among the various types of hypophyseal immunotypes. Despite their well-differentiated nature and their "benign" constitution, an important number of hypophyseal tumors will be surely invasive within the sclerous, osseous and neural structures. Because the aggressive behavior of invasive adenomas is generally resolved by surgical treatment, this aspect of their biology does not perfectly reflect within their histopathological aspect. In fact, the invasive tumors with local extreme aggression are most often deceiving due to their relatively harmless histopathological aspect. The usual morphological signs of tumor aggression, namely pleomorphism, nuclear atypia, hemorrhage, high cellular and mitotic activity, poorly correlate with the invasive potential of pituitary tumors, with their proliferation capacity, their tendency of post-surgical recurrence or with their global biological behavior.

Corresponding author: Mihaela N. Popescu, Lecturer, MD, PhD; e-mail: mihaelavoica123@yahoo.co.uk, elena.ionesc@yahoo.com

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20. Indirect pulp capping in young patients: immunohistological study of pulp-dentin complex

Mihaela Jana Tuculina, Mihaela Raescu, Ionela Teodora Dascalu, Mihaela Popescu, Claudia Florina Andreescu, C. Daguci, C. N. Cumpata, Vanda Roxana Nimigean, Ileana Monica Banita

Indirect capping is a complex therapy exclusively needed in deep cavities that provides, using biomaterials, a disinfection of the dentinary sore and seals the dentinary tubules, protects the pulp of physical mechanisms and chemical agents and stimulates the mechanisms that produce new dentin. Following this idea, we studied the histological changes in the dental pulp tissue and also the specific immunohistochemical response in various structures when an indirect capping technique was used. We used special histological techniques followed by classical staining or by immunohistochemical reaction in order to assess the odontoblastic, and the vascular reaction. The immunohistochemical study allows us to evaluate the changes in the pulp-dentin complex, as the result of the changes in the dentinal tubules permeability and the biological reactions at this level.

Corresponding author: Ionela Teodora Dascalu, Assistant Professor, DDS, PhD; e-mail: marceldascalu@yahoo.com

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21. Histopathological prognostic and risk factors in patients with laryngeal neoplasms

Gh. Iovanescu, Marioara Poenaru, C. Doros, O. Boruga

Head and neck cancer is a public health problem, accounting for the fifth place of all human cancers. Each year in the US, 3% of new cases of cancer develop in this sphere. Histopathology of the primary tumor and/or laterocervical adenopathy is essential to establish the therapeutic strategy. Histopathological findings reveal a diagnostic certainty and also provide prognostic information. Patients and Methods: We have prospectively studied a number of 95 patients with laryngeal neoplasms, which were treated in the ENT Clinic of Timisoara, Romania. The group of patients with neoplasic disease was subdivided into two groups: group 1 - patients with palpable laterocervical adenopathy and detected primary tumor (T+N+), and group 2 - patients without palpable laterocervical adenopathy, but with detected primary tumor (T+N0). Only patients with squamous cell carcinomas were included in this study. We assessed the following microscope parameters: neoplasm confirmation, histological type of neoplasm, the degree of tumor differentiation, intratumoral inflammatory reaction, perineural infiltration and the cell invasion in the small vessels. All patients underwent surgery. Results: N2 and N3 stage adenopathies were more common in patients with medium or low degree of cell differentiation; they were associated with weak inflammatory response of the primary tumor, within vascular invasion alone or associated with the perineural infiltration. N0 and N1 adenopathies were associated with average and good cell differentiation degree, with rich inflammatory infiltrate. Conclusions: Histopathological examination of the primary tumor and cervical adenopathy is essential in establishing the diagnosis and the therapeutic approach. Histopathological examination offers information on the prognosis. Cell differentiation degree, inflammatory reaction and the cell invasion in the small vessels are the main prognostic factors of laryngeal neoplasms.

Corresponding author: Gheorghe Iovanescu, University Assistant, MD, PhD; e-mail: giovanescu@gmail.com

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22. Histological changes of gingival epithelium in diabetic patients with periodontal disease

M. R. Popescu, Mimi Nitu, L. Dragomir, M. Olteanu, Madalina Olteanu

The pathogenesis of the periodontal disease is complex, because it derives from the initiation and maintenance of the chronic inflammatory process by the plenty microbial flora and its numerous bacterial products. The purpose of our study was to make evidence of the histopathological changes occurring in the gingival epithelium in patients with both diabetes mellitus and periodontal disease. Materials and Methods: Our study was carried out on gingival tissue from 68 diabetic patients with specific symptoms of periodontitis. The preparations were processed by Hematoxylin-Eosin staining technique and trichromic staining after the Goldner-Szekely method. Results: Although non-specific, since they have been generally found in chronic gingival inflammation, the noticed changes were significant and some of them had specific aspects for the diabetic patients: gingival epithelium changes like acanthosis type; the presence of inflammatory cells such as polymorphonuclear leukocytes and/or lymphocytes among the cells of the stratified squamous epithelium; surface ulceration areas covered by hematic and fibrino-leukocytary detritus; the epitheliocytes keratinization in the superficial layers. Conclusions: The diabetic patients with periodontitis frequently associate hypertrophy changes of the gingival epithelium with the epithelial permeability alteration and the presence of the intraepithelial inflammatory cells, subsequently leading to degenerative lesions and epithelial ulceration.

Corresponding author: Mihai Raul Popescu, Associate Professor, DMD, PhD; e-mail: popescumihairaul@yahoo.com

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23. Neuroprotective effect of melatonin in experimentally induced hypobaric hypoxia

Corina Vornicescu, Bianca Bosca, Doinita Crisan, Sumaya Yacoob, Nora Stan, Adriana Filip, Alina Sovrea

Melatonin (MEL) is an endogenous neurohormone with many biological functions, including a powerful antioxidant effect. The aim of the present study was to determine whether MEL protects the brain tissue from the oxidative stress induced by hypobaric hypoxia (HH) in vivo. This study was performed on Wistar rats randomly assigned in four groups, according to the pressure conditions and treatment: Group 1: normoxia and placebo; Group 2: HH and placebo; Group 3: normoxia and MEL; and Group 4: HH and MEL. The following aspects were evaluated: cognitive function (space reference and memory), oxidative stress parameters - serum and brain malondialdehyde (MDA) and reduced glutathione (GSH) levels -, and brain tissue macroscopic and microscopic morphological changes. Exposure to oxidative stress results in cognitive dysfunctions and biochemical alterations: significant increase of MDA and reduction of GSH in both serum and brain tissue. The most important morphological changes were observed in Group 2: increased cellularity, loss of pericellular haloes, shrunken neurons with scanty cytoplasm and hyperchromatic, pyknotic or absent nuclei; reactive gliosis, edema and blood-brain barrier alterations could also be observed in some areas. MEL treatment significantly diminished all these effects. Our results suggest that melatonin is a neuroprotective antioxidant both in normoxia and hypobaric hypoxia that can prevent and counteract the deleterious effects of oxidative stress (neuronal death, reactive astrogliosis, memory impairment and cognitive dysfunctions). Dietary supplements containing melatonin might be useful neuroprotective agents for the therapy of hypoxia-induced consequences.

Corresponding author: Alina Sovrea, Associate Professor, MD, PhD; e-mail: a_sovrea@yahoo.com

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24. Assessment of p53 and HER-2/neu genes status and protein products in oral squamous cell carcinomas

Dorina Stoicanescu, Nicoleta Andreescu, Alina Belengeanu, Noemi Meszaros, Marioara Cornianu

Identification of the genes involved in tumor initiation and progression has led to development of new markers and generated targets for new drugs. This study aimed to evaluate p53 and HER-2/neu genes status of and their protein products in oral cancer patients. Tumor specimens from 116 cases diagnosed with oral squamous cell carcinoma were analyzed. P53 and HER-2/neu immunoreactivity were studied. FISH analysis was performed to elucidate p53 and HER-2/neu gene status. Male cases represented 84% of the group. The majority of cases were between 51-60 years and moderately differentiated oral carcinoma had an incidence of 58.6%. Thirty-four cases showed p53 overexpression, negative immunoreaction was observed in 16.37% of cases. 66.38% of cases had p53 deletion, with an increased rate observed in neoplasms of the tongue. Immunohistochemical analysis of HER-2/neu receptor protein revealed that 76.72% were negative, 5.17% had weak immunostaining, 14.65% had +2 score, the others had +3 score. 24.1% of cases were analyzed using FISH technique, of which 25% were without amplification, but with polysomy for chromosome 17. 18.1% of total cases were amplified, with the rate HER-2/neu:CEP17 higher than 2. Of the 77 cases with a single p53 allele, 20 associated HER-2/neu amplification, 31 had positive anti-HER-2/neu immunoreaction, but did not have HER-2/neu:CEP17 rate >2. There was a significant association between HER-2/neu amplification and deletion of a p53 allele. These results could justify more extensive research to assess p53 and HER-2/neu gene status as significant prognostic factors in oral cancers.

Corresponding author: Nicoleta Andreescu, University Assistant, MD, PhD; e-mail: nicollandreescu@yahoo.com

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25. The effect of ultraviolet radiation on the cornea - experimental study

Andreea Golu, Irina Gheorghisor, A. T. Balasoiu, Fl. Balta, E. Osiac, L. Mogoanta, Adriana Bold

Ultraviolet (UV) radiation in high doses may have harmful effects on the eye. The sources of UV radiation are the sun, as well as some artificial sources such as UV lamps or voltaic arcs. Chronic exposure to UV can cause damage to the anterior pole of the eye, ranging from minor (pterygium) to serious photokeratitis. In our study, we applied a UV dose of 6.5 J/cm(2) in the wavelength range of 290-400 nm, for five consecutive days per rat anterior pole of the eye. Seven days after the last dose of radiation, the animals were sacrificed, harvesting both the irradiated and the non-irradiated eye. Histological and immunohistochemical examination of the lesions revealed that the greatest damage to the epithelium was recorded prior to and 2/3 of the remaining corneal stroma. The epithelial lesions we found varied from pseudokeratosis and detachment of the Bowman epithelium membrane to deep epithelial necrosis. Within the corneal stroma, we observed the formation of interstitial edema with disruption of the collagen structure. We also noticed the presence of an inflammatory infiltrate composed mainly of lymphocytes and CD68+ and CD163+ macrophages, as well as the occurrence of vascular devices. These consisted of angiogenesis capillaries with structured wall composed mainly of endothelial CD34+ precursor cells and a basal membrane rich in collagen IV fibers.

Corresponding author: Laurentiu Mogoanta, Professor, MD, PhD; e-mail: laurentiu_mogoanta@yahoo.com

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26. Fibro-hyaline involution of a papillary thyroid carcinoma metastasis in a lymph node, consecutive to radioiodine therapy, mimicking a parathyroid adenoma. A case presentation

Edith Dee, Andrada Loghin, Adela Nechifor-Boila, R. Neagoe, Ionela Pascanu, Angela Borda

Objective: The aim of the study is to present the unusual changes that a lymph node metastasis of papillary thyroid carcinoma (PTC) underwent after radioiodine therapy, leading to the confusion with a parathyroid adenoma (PA). Patient and Methods: Eight years after a total thyroidectomy and radioiodine ablation with 73.35 mCurie 131-I for PTC, a 67-year-old female presented with an enlarged, painless, nodular mass in the left lateral neck region. Clinical examination revealed a firm nodule located on the site of the left inferior parathyroid gland. Elevated serum parathyroid hormone level (120 pg/mL) and parathyroid scintigraphy led to a suspicion of PA. A minimally invasive surgical procedure was performed to remove the mass, which was sent to the Department of Pathology, Emergency County Hospital, Tirgu Mures, Romania, as left PA. It was fixed and processed for microscopic evaluation. Results: On macroscopic examination, the surgical specimen was oval; it had 13 mm at the largest diameter and weighted 2 g. On microscopy, the lesion appeared as a fibro-hyaline, intensely acidophilic, acellular mass, with calcifications. It was limited by a delicate capsule in which one typical psammoma body was present. At the periphery, on one single level, a small mass of cells of indefinite origin was noticed. Immunohistochemistry (IHC) was done to ascertain the origin of these cells: they were negative for Pan-Cytokeratin AE1/AE2, Parathormone and Thyroglobulin antibodies, but positive for Leukocyte Common Antigen (LCA) antibody, proving that they were lymphocytes, most likely residual from a lymph node. Conclusions: These IHC data, together with the microscopic feature, the presence of the psammoma body and the patient's history, excluded a PA and led to a diagnosis of fibro-hyaline involution of a PTC metastasis in a lymph node, consecutive to radioiodine therapy. Without careful microscopic examination and accurate clinical information, this lesion could represent a real diagnostic challenge.

Corresponding author: Andrada Loghin, Lecturer, MD, PhD; e-mail: andradaloghin@yahoo.com

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27. Pseudorheumatoid disability man with chronic tophaceous gout: a case report

Rodica Traistaru, Viorela Enachescu, Diana Kamal, Camelia Foarfa, Otilia Rogoveanu

Gout is a type of inflammatory arthropathy that affects the peripheral joints and results from the accumulation of monosodium urate (MSU) crystals in the synovial fluid and other tissues. This disease is the most common form of inflammatory arthritis in men over 40 years of age. The fundamental biochemical abnormality in gout is an increase in serum urate (SU) concentration. These needle-like crystals induce not only acute episodes of inflammatory process into the surrounding area, but also, in the long-term history of the disease, chronic inflammation that is associated with changes in articular and periarticular structures. The next step caused by deposited MSU crystals is represented by the tophus formation and chronic gouty synovitis. The presence of tophi has been associated with greater physical functional disability in gout patients. We presented a case of severe chronic tophaceous gout in a 48-year-old man with chronic hand arthritis and urolithiasis, to point the significance of complex assessment (clinical, functional, imagistic and histological exams) in the diagnosis of a soft tissue lesion, especially in hands.

Corresponding author: Rodica Traistaru, Lecturer, MD, PhD; e-mail: rodicatraistru@hotmail.com

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28. Histopathological, immunophenotypic and clinical particularities and evolution of a case of hepatosplenic T-cell lymphoma in transformation to leukemia

Erzsebet Benedek Lazar, Judit Beata Kopeczi, Aliz Beata Tunyogi, Eniko Kakucs, Emoke Horvath, M. Turcu, I. Benedek

We present the possibilities of diagnosis correlating the pathological, immunophenotyping and clinical aspects of a rare case of T-cell lymphoma in a 23-year-old patient with leukemic transformation. In our consideration, it is very important to describe this case because in the literature there are very few cases presented and the treatment of this type of lymphoma does not present optimal results, the evolution of the patients being from three months to two years. The treatment modality that gives the possibility to prolong survival and cure is hematopoietic stem cell transplantation.

Corresponding author: Emoke Horvath, Lecturer, MD, PhD; e-mail: horvath_emoke@yahoo.com

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29. A case of non-Hodgkin lymphoma in a patient with chronic myeloid leukemia

Amelia Maria Gaman, Camelia Dobrea, Ionela Rotaru

Chronic myeloid leukemia is a clonal expansion of hematopoietic progenitor cells characterized by exaggerated proliferation of granulocytic lineage, with chronic phase, accelerated phase and blast crisis. Accelerated phase and blast crisis may be associated with extramedulary disease. Extramedullary transformation of CML can be determined both in nodal and extranodal sites. Non-Hodgkin lymphoma is rare in chronic myeloid leukemia and may be misdiagnosed as an extramedullary lymphoid blast transformation; the majorities are T-cell lymphomas with an immature thymic phenotype, while peripheral B-cell lymphomas are rarer. We report the case of a 79-year-old woman carrier Ph+ chronic myeloid leukemia who developed at eight months of diagnosis an accelerated phase of CML associated simultaneous with a tumor of soft palate, which was initial considering an extramedullary disease. The patient was treated with specific chemotherapy for accelerated phase of CML (Cytosinarabinoside) + Anagrelide, and reversed to secondary chronic phase of CML, but soft palate tumor persists. The immunohistochemical findings of bone marrow trephine biopsy examination showed chronic phase of CML (negativity for immature cells such as CD34, Tdt) and the biopsy of soft palate tumor and immunohistochemical findings revealed a primitive non-Hodgkin lymphoma (NHL) with medium B-cells (CD20, CD79a positive) and excluding an extramedullary blast crisis (CD34 negative, Tdt negative). Cytogenetic analysis in tumor revealed absence of Philadelphia chromosome. The patient was treated with local radiotherapy for NHL, with a favorable evolution and Hydroxyurea 1 g/day for CML with hematological remission. A localized lymphoid neoplasm may be an extramedullary localized blast crisis of CML or a distinct malignancy, with distinguished therapy and prognosis. A correct diagnosis based on a complex investigation: immunohistochemistry, conventional cytogenetic analysis and fluorescence in situ hybridization (FISH), molecular analysis (Southern blot and RT-PCR) is necessary. Further studies are required to clarify the pathogenetic relationship between chronic myeloid leukemia and non-Hodgkin lymphomas.

Corresponding author: Amelia Maria Gaman, Associate Professor, MD, PhD; e-mail: gamanamelia@yahoo.com

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30. Insulinoma diagnosed as drug-refractory epilepsy in an adolescent boy: a case report

Emoke Horvath, H. Gozar, Liliana Chira, Iulia Dunca, Eva Kiss, Z. Pavai

Solitary insulinoma is a rare pancreatic tumor in all age groups with an estimated incidence of 1 in 250 000 persons a year. It is even rarely in childhood and mostly shows benign behavior. Cases with uncertain or malignant biology are extremely rare with less than 30 cases described in the literature. Here we report a case of pediatric insulinoma, the first in our department files in the past 20 years, with rapid clinical course following a clinical misdiagnosis as juvenile myoclonic epilepsy, which was complicated with low glucose level (20 mg/dL) and neuroglycopenia. Our case underlines some unusual features of a pediatric insulinomas presented without past medical and family history, after surgery complicated with mental retardation and recurrent epileptiform episodes. Despite the small tumor size, low Ki67 index/mitotic rate and benign immunophenotype marked by positivity for pro-insulin but negativity for beta-HCG, the diagnosis was concluded as insulinoma of uncertain biological behavior due to vascular tumor invasion in agreement with the 2003 WHO Classification for Pancreatic Endocrine Neoplasms. Besides these features, perineural invasion can differentiate insulinomas of uncertain outcome from benign insulin producing tumors. Pediatric insulinomas may present misleading symptoms of epilepsy in neglected cases coming from poor socioeconomic background. Chronic insufficient blood glucose level might contribute to mental retardation and epilepsiform myoconvulsions to be prevented. Differentiation between insulinoma with benign and uncertain behavior is difficult where histological pattern and tumor immunophenotype are less important than the critical morphological parameters. Life long follow-up including regular control of blood glucose and abdominal status of patients are essential for proper assessment of clinical outcome of pediatric insulinoma.

Corresponding author: Emoke Horvath, Lecturer, MD, PhD; e-mail: horvath_emoke@yahoo.com

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31. Erupted composite odontoma associated with malformed teeth - unusual dental aberrations following maxillofacial trauma

M. Y. Padmanabhan, R. K. Pandey, R. Aparna

Background: Tooth development may be hampered following traumatic injuries to maxilla or mandible. The developmental deviations that occur in a tooth bud in the line of a jaw fracture range from agenesis to hamartomatous proliferation. Odontomas are benign odontogenic hard tissue tumors, which are hamartomas characterized by slow growth and non-aggressive behavior. 'Erupted odontoma' is a term used to specifically denote odontomas, which are exposed into the oral cavity. These are rare entities with only 25-30 cases being reported so far in the dental literature. Case Description: This clinical report is pertaining to a rare case of an erupted odontoma in an adolescent patient which was presenting as a hard tissue outgrowth along with an extra-oral sinus in the posterior mandibular region. The surgical excision of the odontoma along with the associated malformed teeth resulted in resolution of pain and sinus discharge associated with this lesion. Conclusions: Erupted odontomas need to be considered in the differential diagnosis of hard tissue projections or outgrowths in the site of past maxillofacial injuries. Surgical excision of these lesions enable resolution of patient symptoms associated with this condition as evidenced in the present case and in other cases cited in the literature. The erupted odontoma attached to malformed molars and presenting with signs and symptoms suggestive of osteomyelitis is a rare combination of developmental aberrations following maxillofacial injury.

Corresponding author: Makkada Yuvaraj Padmanabhan, MDS, PhD; e-mail: dentistpad@rediffmail.com

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32. Cranial sutures and diploae morphology

Claudia Corega, Ligia Vaida, Ioana Tiberia Ilias, D. Bertossi, Ionela Teodora Dascalu

The aim of the study was to assess the normal cranial suture and bone diploae ultrastructural morphology. Two types of sutures from different specimens were collected. The micro-CT scanning provided a three-dimensional view of the sutures at a microscopic level thus allowing the evaluation of the development stage and a rapid analysis evaluation of bone and diploae morphology. In the meantime, the micro-CT is able to generate more slices than the normal histology preserving the analyzed specimens and became one of the most powerful tools in the craniofacial area. The micro-CT analysis generated structure-orientated slices that in conjunction with the histological sections provide a high quality quantitative analysis of all cranial sutures and of the cranial bones diploae.

Corresponding author: Ligia Vaida, Associate Professor, DMD, PhD; e-mail: ligia_vaida@yahoo.com

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33. Orbital exenteration - a salvage procedure?

A. Frunza, D. Slavescu, D. Zamfirescu, Luminita Stanciulescu, Ioana Grintescu, V. Enache, I. Lascar

Orbital exenteration is a procedure performed usually for malignancies. The subsequent reconstructive efforts are directed towards rapid and stable healing but allowing detection of recurrent disease, obliteration of any communication between the orbit and surrounding cavities and above all a good quality of life. The surgical options must be tailored to each patient; we have to achieve first disease control and to compensate a 3D defect as best as we can. In the following, we present a series of three patients with aggressive tumors of the midface, admitted in our clinic in the last four years. For each case, orbital exenteration was performed in order to achieve tumor clearance. The results were good excepting one case (a relapse at three years interval) with a squamous cell carcinoma and perineural invasion.

Corresponding author: Dragos Zamfirescu MD, PhD; e-mail: dragoszamfirescu@gmail.com

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34. Well-differentiated neuroendocrine tumor and osteoporosis: incidental findings?

Catalina Poiana, Mara Carsote, M. C. Neamtu, Elena Taina Avramescu, Florina Vasilescu, Dana Terzea, Diana Paun, Raluca Trifanescu, Rucsandra Danciulescu Miulescu

The neuroendocrine tumors (NETs) have an increased incidence related to the age. Secondary osteoporosis might be found in patients with bone metastases and in those with NETs associated Cushing's disease or primary hyperparathyroidism. Primary osteoporosis might be found in postmenopausal women, but in case with non-metastatic NET as G1 NET it is difficult to establish the NET contribution to the bone loss. We present the case of a 53-year-old female accidentally diagnosed with G1 lung NET after surgery of the tumor. The immunohistochemistry pointed positive reaction for CHROMO, SYN and negative for CK7 and TTF1, and a Ki67 of 1-2% (well-differentiated neuroendocrine tumor). The central Dual X-Ray Absorptiometry (DXA) showed osteoporosis based on a T-score of -3. The patient had normal neuroendocrine markers and she was asymptomatic. She remained so for one year and the only therapy provided was weekly alendronate with adequate vitamin D and calcium supplements. Based on the pathological and immunohistochemistry profile, the low risk NET was diagnosed. We encourage the skeletal status assessment as central DXA in postmenopausal women with NETs, regardless clinical evidence of bone loss. The future will provide more epidemiological and pathogenic connections between the two dynamic fields of medicine as neuroendocrine tumors and osteoporosis.

Corresponding author: Mara Carsote, MD; e-mail: carsote_m@hotmail.com

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35. A rare morphological variant of the first maxillary premolar: a case report

V. Nimigean, Vanda Roxana Nimigean, D. I. Salavastru, Lavinia Butincu

Anatomical variations of the dental root morphology may lead to endodontic treatment failure. Knowledge of root morphology and possible anatomical variations may ensure proper endodontic treatment and long-term success. The present study describes the case of a three-rooted maxillary right first premolar with two buccal roots and one palatal root, each of them having one canal, diagnosed with symptomatic irreversible pulpitis. The treatment was applied according to traditional procedures, and the post-therapeutic evolution was favorable without any further complications.

Corresponding author: Vanda Roxana Nimigean, Assistant Professor, PhD; e-mail: vandanimigean@yahoo.com

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36. Refractory anemia with ringed sideroblasts and thrombocytosis without JAK2 V617F mutation: report of three cases

Aurelia Tatic, Mariana Vasilica, Adriana Colita, Didona Vasilache, Camelia Dobrea, Cerasela Jardan, Amelia Maria Gaman, Ana Manuela Crisan, D. Colita, D. Coriu

In the WHO classification, there is a provisional entity called Myelodysplastic/Myeloproliferative Neoplasm, Unclassifiable (MDS/MPN, U). Refractory anemia with ringed sideroblasts associated with marked thrombocytosis (RARS-T) was included in this category. Recently published studies report a small percentage of patients with RARS-T. Sixty percent of these have JAK2 V617F mutation, which can suggest the coexistence of two pathological conditions (MDS and MPN). In this paper, we analyzed three patients diagnosed with RARS-T in the Department of Hematology, "Fundeni" Clinical Institute, Bucharest, Romania, during the period 2005-2011. The patients were investigated with cytogenetic exam and molecular biology. In these three cases were identified morphological features of multilineage dysplasia (two-lineage dysplasia in two cases and three-lineage dysplasia in one case). In two cases, thrombocytosis was under 1000x10(3)/micro-L and clinical evolution was similar to the myelodysplastic syndrome (transfusion dependent anemia with response to administration of erythropoietin). In the third case, the platelets were over 1000x10(3)/micro-L and with response to the treatment with Hydrea, which improved anemia. JAK2 V617F mutation was not identified in any case. RARS-T remains a provisional entity and requires a complex investigation of patients for the correct diagnosis of these patients. Therapeutic options should be personalized to each case in part because there is not yet a standardized treatment of these patients.

Corresponding author: Aurelia Tatic, Teaching Assistant, MD, PhD; e-mail: auratatic@yahoo.com

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37. Bilateral primary fallopian tube carcinoma: a case report

Andreea Veliscu, B. Marinescu, Lucia Costoiu, Cristina Damian, Casiana Stanescu, Luminita Chiutu, Anca Patrascu, Andrada Loghin

Primary cancer of the fallopian tube is a very rare tumor nowadays, accounting for approximately 0.14-0.3% of all tumors of the female genital tract. From these, bilateral primary cancer is found in less than 25% of all cases. We report here a case of bilateral primary cancer of the fallopian tube in a 48-year-old woman, associating uterine fibromatosis.

Corresponding author: Anca Patrascu, Professor, MD, PhD; e-mail: patrascuanca@yahoo.com

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