ROMANIAN JOURNAL of MORPHOLOGY and EMBRYOLOGY

Vol. 52 No. 1 Suppl., 2011
This supplement was not sponsored by Outside Organizations.

1. Foreword. Research Center for Microscopic Morphology and Immunology Craiova: 10 years of history

Fl. Bogdan

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2. Pathology on the edge of interdisciplinarity. A historical epitome

E. Vollmer, T. Goldmann

Pathology is a bridging discipline that involves both basic and clinical biomedical sciences. In this context, it includes both descriptive and mechanistic approaches, with the final goals of further understanding the anatomic and functional changes and underlying molecular events involved in disease-related processes. Pathology studies mainly comprise macroscopic and microscopic examinations, and involve the visual recognition of different patterns in cells and tissues. In time elapsed to this end, it has adopted a dazzling array of methods and techniques from the most varied fields of natural and engineering sciences in order to optimize tissue analysis, which will be summarized in this article.

Corresponding author: Ekkehard Vollmer, Professor, e-mail: evollmer@fz-borstel.de

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3. Genetic patterns of metalloproteinases and their tissular inhibitors - clinicopathologic and prognostic significance in colorectal cancer

V. Surlin, M. Ioana, I. E. Plesea

Colorectal cancer (CRC) is the third most common cancer in men and second in women. Progression and invasion of colorectal cancer is a multistep process involving multiple interactions between the tumor and the surrounding stroma mediated by many proteins, among them metalloproteinases (MMPs) and tissular inhibitors of metalloproteinases (TIMPs). We aimed to review the correlations between the expression of the MMPs and TIMPs genes and the clinicopathologic variables of the CRC. Levels of MMPs genes expression in colorectal cancer correlate with the depth of invasion, hematogenous and lymphatic metastasis, poor differentiation, Duke's stage and prognosis. Levels of TIMP's genes expression correlate with better prognosis and longer survival. There are also some controversial data explained by the fact that most of the studies addressed one or few MMPs and/or TIMPs. The methods to assess the variance in gene expression were not always the same. The promoter regions of metalloproteinases present many polymorphisms and all have allele-specific effects on regulation of MMP gene transcription. Numerous studies on the association of these polymorphisms with cancer susceptibility have been carried out. Most of the studies addressed one or two polymorphisms and their implications. A meta-analysis is necessary to confirm significant correlations. The heterogenicity of the MMPs and TIMPs genetic patterns generated by different studies on colorectal cancer does not allow us to have an overall correlation with clinicopathologic variables and the prognosis of the disease. Studies that involve many MMPs, TIMPs polymorphisms and their tissular expression would be more valuable to better assess the role of those enzymes in the progression of the disease and may serve as a starting point for selective therapeutic approaches.

Corresponding author: Valeriu Surlin, MD, PhD, e-mail: vsurlin@gmail.com

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4. Extra-genitally located endometriosis

I. Dimitrakoupoulos, Petra Surlin, Garofita Mateescu, Mihaela Hincu, Malina Coman, Emma Gheorghe

Endometriosis is defined by the presence of ectopic and endometrial islands outside the uterus. The significance of its finding is also outlined by its less frequent localizations (intestine, gall bladder, lungs) and by its differentiated symptoms complicating the differential diagnosis. The definitive diagnosis can only be reached through a histopathological examination as the detection of this affliction may lead to severe therapeutic errors (radical resection). The importance of an early diagnosis may help in preventing and delaying the effects of this affliction.

Corresponding author: Mihaela Hincu, Associate Professor, MD, PhD, e-mail: mihaela_hincu10@yahoo.com

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5. Histological and ultrastructural analysis of the seminiferous tubule wall in ageing testis

O. T. Pop, Corina Gabriela Cotoi, I. E. Plesea, Mihaela Gherghiceanu, S. D. Enache, E. Mandache, Gratiela Hortopan, R. M. Plesea

The authors evaluated possible morphological changes of basement membrane (BM) and lamina propria (LP) of seminferous tubule wall (ST) related to ageing. Surgical samples of testicular tissue from 28 cases with orchiectomy for prostate adenocarcinoma were processed for light microscopy and transmission electron microscopy (TEM) examination. Seven age groups (AgGr) between 50 and 80 years were designed. Tissue samples were immunomarked for collagen IV and smooth muscle actin. Images were acquired and measured with a specialized software. Thirty ST were randomly selected, with x40-objective, for each case. Five random determinations for each ST and each parameter were performed. Mean values/tubule, case and AgGr were calculated for each parameter. Regression line (RL), slope and significance test for slope were determined for each parameter correlation with ageing. BM mean value was around 0.5 micro-m, with narrow limits of ranging in AgGr but more extended individual limits. RL showed discrete decreasing trend with ageing but without an obvious statistical correlation. LP mean value was around 6 micro-m, also with narrow limits of ranging in AgGr and more extended individual limits. RL decreased discretely with ageing but without an obvious statistical correlation. TEM showed more prominent BM material and more collagen fibers and less fibroblasts in LP of older AgGr and higher fibroblasts density in LP of younger AgGr. Our results showed that BM thickness is apparently decreasing with ageing whereas LP presents extremely variable degenerative changes, with a "mosaic", focal distribution and no tendency to advance with ageing.

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

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6. The role of endoscopic examination accompanied by histological examination on biopsy samples in the diagnosis of gastric carcinoma

D. Ghita, Aurelia Glavici, A. Saftoiu, I. E. Plesea, S. Cazacu, Claudia Georgescu, T. Ciurea

Introduction. Endoscopic evaluation is one of the most important explorations in the diagnosis of gastric cancer, increasing its value by adding biopsy sampling and histopathologic examination, especially in early forms of gastric malignant proliferations. The aim of this study was to evaluate some descriptive parameters of macroscopic and microscopic aspects of gastric carcinomas defined with the help of endoscopic investigation and gastric biopsies sampled during endoscopic examination, and their correlation with patient survival. Materials and Methods. The study was performed on a group of 119 patients diagnosed with gastric carcinoma. The study material was represented by: tissue fragments obtained by endoscopic biopsy, clinical observation charts, histopathologic diagnosis records, ledgers for records of endoscopic investigation, and endoscopic images recorded for each patient. Biopsies were taken preferentially from areas with the highest risk of malignancy. Tumor fragments were subjected to conventional histological processing techniques (fixation and inclusion in paraffin) and then were stained with HIM. The study included two chapters: the imagistic study and the pathologic study. The parameters assessed were: the site of the lesion within the stomach, the endoscope macroscopic appearance, the microscopic appearance of the endoscope biopsy sample, and the survival, followed up until 48 months. Results. The most frequently observed macroscopic aspect was the fungating one, in more than two thirds of all cases, followed by the infiltrating one, but all with a poor survival - about 25-30% at 24 months. The most frequent location was the antro-pyloric region, with the highest survival rate of 25% at 24 months, followed by the gastric corpus and the lesser curvature. Most of the tumors were restricted to a single segment of the stomach. The most frequent microscopic aspect was the tubular one (75% of all cases) with its poorly differentiated variant (39 of the 91 cases). The 24 months survival was under 50% for all morphological types of carcinoma, with the lowest ones in the tubular and mucinous types (around 25%). The secretory pattern was identified on biopsy samples stained with HE in only 15% of the cases, and did not influence the survival of the patients. Conclusions. The combined histologic and endoscopic morphologic investigation allowed the shaping of an accurate morphologic and prognostic preoperative profile in gastric carcinomas.

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

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7. Morphoclinical study of intracerebral hemorrhage with subarachnoid effusion

Carmen Corina Stancu, I. E. Plesea, S. D. Enache, R. Diaconescu, A. Camenita, Mihaela Tenovici

Aim. The study is an integrated assessment of clinical, imagistic and morphological parameters in severe intracerebral hemorrhages (ICH) complicated with subarachnoid effusion (SAE). Materials and Methods. The studied group had 37 cases of patients with ICH and SAE who were hospitalized in the Emergency County Hospital of Craiova and died during hospitalization. The parameters evaluated were clinical (relation with the seasons, age, sex, arterial blood pressure, the motor deficit, degree of coma and Glasgow score at admission) and morphological (the sites of the intraparenchymal hematoma and SAE, the size of the intraparenchymal hematoma, the presence of the mass effect, and the association of intraventricular effusion). The latter were assessed on CT films and during autopsy. Results. The presence of SAE as a complication of ICH showed a predilection for cold seasons, especially winter. From the 37 studied cases, 18 were men and 19 women. 51.3% of the patients were in the fourth and fifth life decade. Almost 73% of the patients had third stage arterial hypertension at admission, over 56% motor deficits and almost 49% Glasgow scores lower than 6. The hematomas had huge dimensions as compared to hosting encephalic structures, in lobar sites involving more than one lobe. Other risk factors as mass effect, perilesional edema and intraventricular extension (IVE) were constantly present. Conclusions. The association of SAE with other independent risk factors such as hypertension, low Glasgow scores, dimensions of hemorrhagic foci, presence of mass effect, perilesional edema and intraventricular extension (IVE) results in the death of patient despite any sustained therapeutical intervention.

Corresponding author: Carmen Corina Stancu, PhD, e-mail: doctorstancu@yahoo.com

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8. Preliminary study of bipolar hip prosthesis - influence of acetabular bone interactions on bone morphology

I. E. Plesea, D. Anusca, Simona Bondari, O. T. Pop, F. Poenaru, V. Dascalu, Mirela Ghilusi

Periprosthetic bone changes following hip arthroplasty are yet to be completely described. The material consisted of imagistic records (X-ray films, CT and MRI scans) and of acetabular bone tissue sampled from 14 cases with femoral head prosthesis and revision of the prosthesis fixed and decalcified in Duboscq-Brazil solution and stained with Hematoxylin and Eosin, trichrome van Gieson and trichrome Masson. Acetabular bone is home of a great variety of morphological changes that can be divided in degenerative and regenerative changes seen in both compact and trabecular components but only inside the maximal pressure area of the acetabular roof. Our preliminary morphological study revealed the existence of an adaptation effort to the mechanical stress materialized through a dynamic process of bone remodeling in the maximal pressure area.

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

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9. Pleurisies - the experience of "Tudor Vladimirescu" Hospital of Pneumology II: morphological study

M. Olaru, I. E. Plesea, Iulia Capitanescu, Daniela Dragnei, B. Stanoiu, Fl. Bogdan

Pleural effusions are still representing a challenge in daily practice. Materials and Methods. This retrospective study on 221 patients with pleurisies hospitalized in our unit is focused on the contribution of different types of pleural fluid morphological evaluation in setting a correct etiological diagnosis. The algorithm of investigation included: gross aspects assessment on X-ray records and by direct observation of pleural liquid obtained by thoracentesis and microscopic assessment on cytology slides of pleural fluid and on histopathological samples obtained by pleural needle biopsies. Results. Mycobacterial etiology was the most frequent, with 72% of all cases, followed by tumoral etiology. Cytologic examination of pleural fluid was useful in establishing the final diagnosis in 66.1% of cases, histopathological assessment being imposed for the rest of cases. Discussion. Imagistic investigation offered appropriate information concerning the site and extention of pleural effusions and guided, in certain cases, the needle biopsy. Gross aspect of pleural fluid oriented quite well the suspicion diagnosis. The use of a set of cytological "formulas" was useful in filtering subsequently the suspicion diagnosis. Histopatholgical examination of pleural tissue samples established the final diagnosis in cases where etiology was still uncertain after laboratory and cytological examination and subtyped further the pathologic processes within each main category of etiology. Conclusions. A correct diagnosis of pleural effusions could be achieved only by going through a precise algorithm of investigation where, besides thorough clinical examination and laboratory tests especially of pleural liquid, morphological assessment and in particular cytologic examination of pleural liquid and histopatological examination of pleural tissue samples are essential.

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

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10. The importance of monitoring protocols in cervical carcinoma screening

Madalina Bosoteanu, C. Bosoteanu, Mariana Deacu, Mariana Aschie

Romania has supremacy in terms of European statistical indicators of cervical cancer, a fact attested by the studies made by international organizations. The present study is based on cytological evaluation of a group of 9269 cervico-vaginal smears, segregated in various groups that were monitored by standard diagnostic and therapeutic protocols, attitude based on an excellent collaboration with the gynecologist. This cooperation led to the elaboration of a set of protocols for follow-up of patients assessed by Babes-Papanicolaou test, in order to assure an adequate management for all cervical lesions. An important feature of this study is that histopathological examination of cases cytologically designated as HSIL showed, along with changes of HSIL-CIN2 and HSIL-CIN3, also carcinoma in situ and invasive squamous cell carcinoma lesions, emphasizing the importance of the pathologic diagnosis of certainty. This idea is also supported in cases of glandular cell atypia, whose microscopic evaluation identified premalignant and malignant lesions, both in endometrial and endocervical site. A particular aspect of the analyzed batch consists in the description of a subgroup of false-negative cytodiagnostic results associated with cervical carcinoma, highlighting the causes and the possibilities to avoid further errors. Overall analysis of results reveals major involvement of the pathologist in providing the sequence from cytology to histopathological diagnosis and to establish diagnostic continuity.

Corresponding author: Madalina Bosoteanu, Lecturer, MD, PhD, e-mail: mbosoteanu@yahoo.com

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11. Nuclear comparative morphometric study between DCIS and normal resting mammary gland tissue

Mariana Deacu, Mariana Aschie, Madalina Bosoteanu, L. Petcu

Ductal carcinoma in situ (DCIS) is the malignant epithelial cell proliferation that affect only ducts, including lobular, without basement membrane interruption. Benign or malignant cell phenotype is defined by nuclear appearance. Morphometric analysis could provide quantitative information about nuclear profile in several lesions. In this study, we assess nuclear morphometric features of mammary epithelial cells in DCIS compared to normal resting mammary gland tissue. For morphological evaluation, we included two groups of mammary gland tissue. The first group comprised breast tissue from 20 women surgically treated and histopathologically confirmed with DCIS. The second control group was represented by normal resting mammary tissue obtained from another 20 women surgically treated for fibroadenoma. Evaluated morphometric parameters were: nuclear area (NA), nuclear perimeter (P), maximum diameter (Dmax), minimum diameter (Dmin), elongation factor (E). Morphometric assessment of DCIS nuclei showed significant higher values than normal resting breast tissue. Morphometric analysis gave information about tumor aggressiveness, invasion tendency and disease prognostic.

Corresponding author: Mariana Deacu, Lecturer, MD, PhD, e-mail: deacu_mariana@yahoo.com

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12. Vascular endothelial growth factor (VEGF) expression in the lung in toxic septic shock

Irina Manoilescu, S. Teleman, Elena Cojocaru, Doina Mihaila, P. Plamadeala

Introduction: The need for reasoning with medical evidence the different types of shock, especially when there are medical and legal implications, has determined the search of biological markers of the shock. In the case of toxic septic shock, the most important markers to be used are: the cytokines, the tumor necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6), procalcitonin, lactoferin and the vascular endothelial growth factor (VEGF). VEGF has an essential role in angiogenesis and vascular permeability. Materials and Methods: In our study group, we included 30 cases of different types of shock in which we studied the VEGF expression in the lungs. We added also 10 fragments of lung as control group. According to the etiology, the 30 cases of shock were: 15 with a toxic septic shock and 15 with a hemorrhagic shock. In all these cases we used the classical Hematoxylin and Eosin staining method and the immunohistochemical reactions for VEGF-A. Statistical analysis was performed using SPSS 13.0. Results: The VEGF expression was decreased in all the cases of toxic septic shock, in the endothelium and also in the alveolar epithelium, compared to a high level of expression in other cases of shock and in the control lung. Conclusions: These data allow us to appreciate that VEGF has a different expression in different types of shock and in the normal lung. We observed a statistically significant difference between VEGF expression in toxic septic shock and hemorrhagic shock (p=0.000001). There is a similarity of VEGF expression between hemorrhagic shock and the control lungs (p=0.00001). An obviously low VEGF expression in the toxic septic shock represents a useful biological marker in the forensic medical cases.

Corresponding author: Petru Plamadeala, MD, PhD, e-mail: p_petru@yahoo.com

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13. Immunohistochemical nuclear staining for p53, PCNA, Ki-67 and bcl-2 in different histologic variants of basal cell carcinoma

Claudia Mateoiu, A. Pirici, Fl. Bogdan

Background: Basal cell carcinoma is the most common form of human cancer. Increased expression of p53 has been found in the majority of basal cell carcinomas (BCCs); however, UV-light-induced signature mutations are present in only about 50% of cases. Increased nuclear staining with an immunohistochemical marker of proliferation and apoptosis has been correlated with aggressive behavior in BCC. Objective: Our purpose was to correlate markers expression of apoptosis (p53 and bcl-2) and cell proliferation (Ki-67 and PCNA) with histological indicators of tumor severity. Methods: We used immunohistochemical stains for p53, PCNA, and Ki-67, in superficial, nodular and sclerosing BCC, to determine whether the staining patterns differ in these different histologic variants of BCC. Results: Bcl-2 expression was significant in basal cell carcinomas said to be aggressive (morpheaform and nodular types). Of the studied tumors, 66.7% (n=14) strongly expressed p53. Our results show a greater expression of Ki-67 in nodular and superficial basal cell carcinoma. PCNA showed a strong expression in all types of tumors. Conclusion: Studies employing molecular and genetic biology techniques, associated with histomorphology, lead to the identification of risk factors in the development of more recurring and aggressive lesions.

Corresponding author: Claudia Mateoiu, MD, PhD student, e-mail: claudiamateoiu@gmail.com

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14. The histopathological study of osteoporosis

Fl. Marcu, Fl. Bogdan, Gabriela Mutiu, L. Lazar

The osteoporosis is characterized by the imbalance between the activity of the osteoblasts, the bone forming cells, and the osteoclasts, the cells that resorb the bone tissue, imbalance that favors the osteoclasts. As a conclusion, in the case of osteoporosis, for the same volume, the bone is less compact and more fragile. The objective of our study is to make a histological evaluation of the different elements of the bone tissue in many 47 bone samples: 27 bone fragments were collected from the head and the femoral head of patients who required hip arthroplasty and 20 bone fragments were collected from the vertebral body of dead patients. The results of our study emphasized the thinned trabeculae of the bone that lost continuity, the preferential resorption of the horizontal trabeculae, the consecutive trabecular anisotropy and the reduction of the trabecular connectivity with enlarged areolae and the adipose degeneration of the marrow. One notices in the osteoporosis a reduction of the trabecular network connectivity directly proportional with the stage of the illness; thus, we determined a strong reduction of the trabecular connectivity in advanced osteoporosis stages. The growth aspects of the medular adiposity, associated with the intratrabecular connectivity concurs to highlight the functional connection between bone and marrow. The diminution of the medullar cellularity together with its enrichment in fat cells has negative outcomes on the bone.

Corresponding author: Florin Marcu, Assistant Professor, e-mail: mfmihai27@yahoo.com

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15. Imagistic and histopathologic concordances in degenerative lesions of intervertebral disks

Mariana Cevei, Elena Rosca, L. Liviu, Gabriela Mutiu, Dorina Stoicanescu, Liliana Vasile

Lumbar disk lesions in 47 cases were initially diagnosed using MRI investigation, then, after surgery, biological and histopathological aspects of intervertebral disks were revealed. Pieces from intervertebral disks were used for electron microscopy studies in order to determine collagen in the components of the intervertebral disk. The aim of the present study was to highlight the correspondence between the MRI aspect in cases with clinically manifest lumbar hernia, staged according to MRI Modic classification, and the histopathological aspect in patients with surgical interventions on the intervertebral disks. 4/5 of the analyzed disks had advanced forms of degenerescence of the intervertebral disks: hyalinized disk cartilage +/- intradiskal calcification or ossification zones, chronic inflammatory infiltrate at the disk cartilage level. Electron microscopy studies made on disk fragments obtained by discectomy revealed quantitative and qualitative changes of all types of collagen at the level of the three anatomical structures of the intervertebral disks, which correspond to the MRI changes.

Corresponding author: Mariana Cevei, Associate Professor, MD, PhD, e-mail: cevei_mariana@yahoo.com

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16. The study of synovities with articular inflammatory liquid, through clinical-statistical, histological and immunohistochemical methods

Oana Pobirci, Fl. Bogdan, D. D. Pobirci, Elena Rosca, Carmen Adina Petcu

Introduction: The examination of the synovial is very useful in the positive and differential diagnostic of many articular diseases and especially in the conditions of acute monoarthritis. Materials and Methods: The study focused on the establishment of clinical-statistical, histopathological and immunohistochemical correlations on a group of cases anatomo-pathologic diagnosed with synovity with articular inflammatory liquid. The group was divided in five subgroups: rheumatoid polyarthritis, uric arthropathy (gout), TBC arthritis, sarcoidosis and villo-nodular synovity. Results and Discussion: During the clinical-statistical study the number of arthritis with articular inflammatory infiltration was pursued, the specific location of them and the correlation of the clinical dates with paraclinical ones. In the histopatological and immunohistochemical analysis was pursued the presence of the inflammatory infiltration through the implication of both types of B- and T-lymphocytes in different proportions taking into consideration the cause of the synovity. Conclusions: The synovial biopsy is indicated at patients at whom the diagnostic is not established after the clinical evaluation. The examination of the synovial tissue can be the only way of establishing a definitive diagnostic in inflammatory arthropathies.

Corresponding author: Florin Bogdan, Professor, MD, PhD, e-mail: rjmorphembryol@yahoo.com

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17. Correlations between intralobular interstitial morphological changes and epithelial changes in ageing testis

O. T. Pop, Corina Gabriela Cotoi, I. E. Plesea, S. D. Enache, Florina Carmen Popescu, M. A. Enache, R. M. Plesea

The study focuses on the possible influences of intra (I) lobular (L) stromal compounds [intertubular spaces and seminiferous (S) tubule (T) wall (W)] morphologic changes on S epithelium (E) during ageing process. The material consisted of surgical samples of testicular tissue from 192 patients with orchidectomy for prostate carcinoma. Seven age groups were designed, from 50 to 80 years. Tissue samples were fixed in neutral buffered formalin, embedded in paraffin stained with HE, Goldner and Gomori and immunomarked (in a subgroup of 28 cases) for smooth muscle actin, collagen IV, and CD34. SE had an uneven involution, both individually and inter-individually, but with normal spermatogenesis in many of ST. E degenerative changes were seen mainly in L periphery. Different stages of maturation arresting were more frequent in older patients. IL septae had changes with extremely variable intensity, dispersed mainly in L periphery, without significant spread and without extensive trend with ageing. Leydig cells showed focal hyperplasia without extensive trend related with ageing. STW presented strictly in the internal layer of lamina propria (apposed to basement membrane of ES) a focal sclerosis, with variable extension concerning its presence, thickness and T circumference (T without sclerosis, with focal sclerosis and with fibro-hyaline "collar" - FHyC) but not related with ageing. IL arteriolae showed focal areas of degeneration with a wide individual and inter-individual range of intensity and extension, but not related with age. Capillary network (CN), with both its peri-T and intramural segments, was present in all age groups, with no quantitative endothelial changes and decreasing only in very old cases. FHyC was often associated with E atrophy. STW focal sclerosis could explain focal degeneration of SE in senescence, although CN undergoes no significant changes.

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

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18. Invasion assessment in gastric carcinoma - imagistic and histopathologic combined study

D. Ghita, Aurelia Glavici, I. E. Plesea, A. Saftoiu, Daniela Dumitrescu, T. Ciurea

Introduction. Imagistic examinations have an essential role in the assessment of gastric cancer both in earlier and advanced stages. Among these, endoscopic ultrasound (EUS) especially contributes to the diagnosis by direct visualization of the tumor and to the staging and prognosis assessment through the evaluation of the gastric wall and regional lymph nodes. Computer tomography (CT) examination also has an important role, especially in advanced stages, in the assessment of the prognosis, contributing to TNM staging. The aim of this study is a comparative assessment of tumor invasion degree by these two imagistic methods and the correlation of the imagistic diagnosis with histopathologic assessment results on surgical specimens. Materials and Methods. The basis of the study was represented by a group of 38 patients with gastric carcinoma investigated both by endoscopic ultrasound and computer tomography from which 15 cases with surgical excision of the tumor followed by microscopic examination were selected. Studied material was represented by: patientsâ?? medical charts, registers for ultrasound endoscopy and CT investigation, endoscopic and CT images, surgical excision samples and pathology reports. Tumor fragments were processed by classical histological techniques (fixation and paraffin wax embedding) and staining (HE). Results. Comparing the CT examination with ultrasound endoscopy results showed that CT examination overestimated the invasion grade of the gastric wall (T) but accurately defined the grade of lymph node invasion (N) and metastases (M). Comparing the results of ultrasound endoscopic examination with those from histopathological examination showed that the first method underestimated the grade of invasion of the gastric wall (T) while the latter correctly defined the grade of lymph node invasion (N) and metastases (M). Comparing CT examination results with histopathological ones showed that CT overestimated both the grade of invasion of the gastric wall (T), lymph node invasion (N) and metastases (M). Conclusions. In the preoperative assessment of the invasion stage of gastric carcinoma (TNM), ultrasound endoscopy is the elective imagistic investigation for predicting the grade of invasion of the gastric wall while CT examination is a more accurate assessment of lymph node extension and metastases, the precise definition of invasion stage being only the result of histopathological examination on surgical specimens.

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

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19. Predictive preoperatory variables of the prostate tumor volume

Andrada Loghin, O. Preda, V. Bacarea, C. Moldovan, D. Porav-Hodade, Alis Dema, Nicole Berger, Angela Borda

Prostate cancer (PCa) is the second most frequent malignant tumor in men worldwide and the most common form of cancer in men over 50-year-old. The adequate preoperative estimation of tumor volume in order to identify small tumors that lack a short-term aggressive behavior and do not necessitate a forthwith-radical prostatectomy (RP) is the subject of various recent studies and numerous debates. In this study, that included 128 cases, we attempted to evaluate some of the common preoperative variables (patient's age, total prostate volume determined on ultrasound examination, serum PSA, the number of positive biopsies and tumor size, the percentage of tumor length and the Gleason score) that could predict the tumor volume on the final RP. Based on these correlations, we develop a scoring system that combines only the Gleason score, the number of positive biopsies and the percentage of tumor length and that has been statistically proved to be correlated and predictive for the tumor volume. Our study brings additional and practical information about a true and effective prospective evaluation of the volume of the PCa.

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

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20. The significance of angiogenesis and tumoral proliferation in renal cell carcinoma

Daniela Puscasiu, Carmen Tatu, R. F. Tatu, Elena Potencz, Roxana Popescu, Ioana Muntean, Doina Verdes

Angiogenesis represents one of the most important factors of the tumor proliferation. Renal carcinoma with clear cells is highly vascularized. Knowing numerous quantification systems of tumor angiogenesis, we used a simple one, the evaluation of the relative vascular density. We studied 61 cases with partial or total nephrectomy performed in the Urology Department of Timisoara County Hospital. We correlated the intensity of angiogenesis with a tumor proliferation factor PCNA (proliferating cell nuclear antigen) and with the monoclonal antibody PC10. Correlation of the two immunohistochemical methods with the degree of the tumor differentiation suggested an inverse ratio between vascular density and tumor proliferation degree.

Corresponding author: Daniela Puscasiu, Assistant Professor, MD, PhD, e-mail: danielapuscsiu@yahoo.com

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21. Prognostic factors in colorectal cancer evolution

V. Strambu, C. Iorga, P. Radu, S. Stoian, C. Puscu, M. Bratucu, D. Garofil, Fl. Popa

Background. This paper tries to evaluate prognostic value of various pre and post-operative colorectal cancer markers. Materials and Methods. In the study conducted in our clinic over a period of five years we tried to emphasize the biological factors of prognostic value in colorectal cancer, and to demonstrate the important role of these factors in predicting survival, but also of early relapse or, in some cases, resistance to chemotherapy. Most important component of these factors remains molecular tumor markers. Results. Of the markers of tumor load increased preoperative serum levels of carcinoembryonic antigen (CEA) means increased risk of neoplastic recurrence and reducing survival expectancy. Aneuploidy tumor cells would have the same importance. Conclusions. Although for their study modern and expensive techniques are necessary, molecular tumor markers have an increasingly role appreciated by researchers both in estimating the risk of relapse and neoplastic dissemination and the response rate to adjuvant treatment. It is estimated that the study of molecular/genetic profile of colorectal tumors in the future will dictate therapeutic decisions ahead.

Corresponding author: Victor Dan Eugen Strambu, MD, PhD, e-mail: drstrambu@gmail.com

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22. Sentinel lymph node study in colorectal cancer using serial sectioning and Hematoxylin-Eosin staining: importance and limitations

I. D. Vilcea, I. Vasile, C. S. Mirea, C. Mesina, S. D. Enache, Mihaela Tenovici, S. Mogoanta, C. Ghita

The lymph node involvement represents an important predictor for survival in colorectal cancer; consequently, the best pathologic evaluation is necessary in order to adequately assess the lymph node status. This study aims to evaluate the impact of sentinel lymph node technique in colorectal cancer in lymphatic basin staging. The study included 43 consecutive operated cases, in which the identification of sentinel lymph node was performed during surgery (in vivo procedure - colon cancer) or immediately after the removal of the resection specimen (ex vivo procedure - rectal cancer). These cases were matched with 45 control cases. The identified sentinel lymph node was separately examined using multiple sections and Hematoxylin-Eosin staining method. The detection rate, accuracy, sensitivity and false negative rate were better for colon cancer (86.36%; 84.21%; 66.66%; 23.07%) vs. rectal cancer (61.9%; 84.61%; 50%; 18.18%), but there are no arguments for the feasibility of the technique in every day practice. Further studies and methods are mandatory in order to improve the staging of the pN status in colon and rectal cancer.

Corresponding author: Ionica Daniel Vilcea, Assistant Professor, MD, PhD, e-mail: dany_vilcea@yahoo.com

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23. Study of malignant fibrous histiocytoma: clinical, statistic and histopatological interrelation

D. D. Pobirci, Fl. Bogdan, Oana Pobirci, Carmen Adina Petcu, Elena Rosca

Introduction: Malignant fibrous histiocytoma (MFH) is a malignant tumor of soft parts observed at approximately 70% of cases situated at the level of the outsides. Patients and Methods: Our study is formed of 14 cases of patients in a period of three years (2007-2010), diagnosed anatomo-pathological with malignant fibrous histiocytoma at the County Hospital of Oradea. The histopathological examination together with the immunohistochemical one is of vital importance in the establishment of the positive diagnostic and furthermore in the establishment of the differential diagnostic with carcinomas, plasmacytomas, osteosarcomas, fibrosarcomas and lymphosarcomas, fact that arouse the doctors to administrate the correct treatment to a specific patient. Discussion: MFH is a tumor of late adulthood that occurs in men more commonly than women. Computer tomography and MRI have been widely used in the diagnosis and staging of MFH. MFH is secondary to another process such as radiation, surgery, fracture, osteonecrosis, Paget's disease, non-ossifying fibroma or fibrous dysplasia 20% of the time. MFH arising from a previous abnormality is usually more aggressive and has a poorer prognosis than primary MFH. Primary osseous MFH is a central lesion found in the diaphysis or metaphysis of the bone that causes aggressive bone destruction and a soft tissue mass. The most common sites in order are the distal femur, proximal tibia, proximal femur and proximal humerus. Primary osseous MFH is less common. MFH is found in the extremities 70-75% of the time and 50% of all cases are in the lower extremity. Other less common sites include the retroperitoneum, and the head and neck. In our study, of 14 patients with malignant fibrous histiocytoma, the highest incidence is during the sixth decade of life and there is a male to female ratio of 8 to 6. In the specialty literature, malignant fibrous histiocytoma tend to occur in children and teenagers but can also occur in older adults as secondary lesions in bone infarcts and radiation fields. This tumor is clinically similar to osteosarcoma and fibrosarcoma, although malignant fibrous histiocytomas have been classified as different from the osteosarcoma group because of a different histology (no tumor bone production). Treatment is similar to that of osteosarcoma. Conclusions: During our study, the average age was of 61 years in comparison with the specialty literature where the average age was of 50 years. The same as in the specialty literature the cases of fibrous malignant histiocytoma studied are more frequently present at men. Regarding the situation, our study shows the affectation of the long bones especially the femur followed by the radius. The histopathological examination together with the immunohistochemical one is of vital importance in the establishment of the positive diagnostic and furthermore in the establishment of the differential diagnostic with carcinomas, plasmacytomas and lymphosarcomas.

Corresponding author: Florin Bogdan, Professor, MD, PhD, e-mail: rjmorphembryol@yahoo.com

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24. Study of clinical-morphological correlations in carcinoid tumors of the colon

Carmen Adina Petcu, V. Surlin, Fl. Bogdan

Introduction: The incidence of carcinoid tumors of the digestive tract increased in the last 30 years. The aim of the study is to find clinical and morphologic correlations in cases with digestive carcinoid tumors. Materials and Methods: A retrospective study on 76 cases with digestive carcinoid tumors divided in five groups according to their location on the segments of the digestive tract. Results: The subgroup of colon carcinoid is analyzed and clinical-morphological correlations are obtained between the demographic data of the patients (age, gender and environment), tumor location, clinical expression and morphologic features. Conclusions: Due to the newly discovered techniques of investigation, the incidence of digestive carcinoid tumors has risen in association with other pathologies for which we make these investigations

Corresponding author: Valeriu Surlin, MD, PhD, e-mail: vsurlin@gmail.com

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25. Cytologic and histopathologic diagnosis in bronchopulmonary squamous cell carcinoma

Ana-Maria Dragan, Elena Rosca, Gabriela Mutiu

The classification of squamous cell carcinomas, based on cellular differentiation features, includes the poorly differentiated epidermoid carcinomas and well-differentiated epidermoid carcinomas. The histogenetic cytologic data clarify conventional cytodiagnosis of poorly differentiated epidermoid carcinomas and poorly differentiated adenocarcinoma, and also eliminate other categories such as large cell carcinoma and small cell anaplastic carcinoma. We conducted a study for evaluating the degree of differentiation of pulmonary squamous cell carcinoma in 620 patients - 551 men (88.8%) and 69 women (11.1%) who had lung cancer confirmed by cytologic, histologic and bronchoscopic examination. The cytologic examination was performed on slides with samples obtained by bronchial brushing and prints of bronchial biopsy stained with the Giemsa method. Histopathologic examination was performed on samples obtained by bronchial biopsy and stained with Hematoxylin-Eosin. At cytologic examination we found: poorly differentiated epidermiod carcinomas in 66 cases (33.8%), moderately differentiated epidermoid carcinomas in 22 cases (11.2%) and well differentiated epidermoid carcinomas in 107 cases (54.8%). Histological examination revealed: poorly differentiated epidermoid carcinomas in 133 cases (45.7%), moderately differentiated epidermoid carcinomas in 32 cases (10.9%), and well differentiated epidermoid carcinomas in 126 cases (43.2%). Our results suggested the importance of the association between cytologic and histopatologic examinations in the diagnosis of lung cancer.

Corresponding author: Ana-Maria Dragan, Associate Professor, MD, PhD, e-mail: dragan_21_noiembrie@yahoo.com

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26. Connections between renovascular hypertension and nephrectomy

Daniela Rahota, Elena Rosca, Gabriela Mutiu

Renovascular hypertension (RVH) is very often an under-diagnosed condition, being discovered incidentally, at a routine medical check-up or during a medical examination for another disease. A number of 217 biological samples (kidney and renal pelvis) resulting from the nephrectomies performed during the period 2008-2009 in the Urology Clinic of Oradea County Hospital have been taken into study. These samples were processed in the Pathology Laboratory of the same hospital. The distribution of nephrectomies according to the urological diagnosis has shown that 38.2% of the patients underwent surgical procedure for renal parenchymal malignant tumor (Grawitz tumor). Out of the 83 nephrectomies caused by renal tumor, in eight cases, benign and malignant angiosclerosis histological changes were seen, which demonstrates a long evolution of hypertension. Its clinical symptoms were masked by those triggered by the tumor. Histopathological changes of benign nephroangiosclerosis with arteriolar vasospasm associated with edema of the wall and/or hypertrophy of the intima with the formation of hyaline deposits were found in three nephrectomies due to hydronephrosis. The pathological study carried on samples obtained from nephrectomies showed that the symptoms of RVH are hidden or accompanied by clinical manifestations of a surgical disease. The percentage of 8.8% of the patients that had histopathologic changes or malignant of benign nephroangio-sclerosis and suffered nephrectomies for a surgical disease is quite small compared to the large number of nephrectomies. It is imperative to identify in due time the secondary mechanism of hypertension in order to perform the surgical intervention which saves the kidney or prevents the extension to other organs. Although no special investigations were performed showing a possible preoperative RVH, it can be stated that the renal origin of the arterial hypertension can be established retrospectively, after nephrectomy. The monitoring of blood pressure values during several years is extremely important for establishing the etiology of renal hypertension.

Corresponding author: Daniela Rahota, Lecturer, MD, PhD, e-mail: dr.rahota@gmail.com

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27. Histoenzymatic and immunohistochemical study during antiangiogenetic treatment in breast cancer cells

Annamaria Pallag, A. Ardelean, Elena Rosca, O. Pop, Ramona Suciu

Breast cancer is the second most frequent cause of death by cancer. In Romania, its incidence is around 4200 new cases per year and the mortality is around 2500 cases per year. During the past few years a great number of anti-neoplastic therapies with higher specificity and efficiency were developed. Monitoring the effects of these polychemotherapies is of great importance together with the study of their influence on cellular metabolism. The purpose of the study was to establish the changes in the cellular energy metabolism and apoptotic potential in the anti-neoplastic therapy with bevacizumab. The results obtained show that the apoptotic potential of malignant cells increases significantly during the anti-angiogenic treatment, with reduction of tumor vasculature and that the mitochondrial apoptotic pathway is activated by releasing cytochrome c from the mitochondrial inter-membrane space.

Corresponding author: Annamaria Pallag, e-mail: annamariapallag@gmail.com

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28. Evidence for the involvement of TGF-beta1-CTGF axis in liver fibrogenesis secondary to hepatic viral infection

Daniela Tache, Fl. Bogdan, Catalina Pisoschi, Monica Banita, Camelia Stanciulescu, Ana Maria Fusaru, Violeta Comanescu

Liver fibrosis is a nonspecific response to injuries, which implies the synthesis of an abnormal extracellular matrix (ECM). TGF-beta (transforming growth factor beta) is a cytokine involved in regulation of several important processes: cell development and differentiation, apoptosis, synthesis and degradation of ECM. CTGF (connective tissue growth factor) is a cysteine rich peptide that belongs to the CCN family of proteins and plays an essential role in the formation of blood vessels, bone and connective tissue. The purpose of this study was to assess TGF-beta1 and CTGF immunohistochemical expression in different stages of liver fibrosis secondary to chronic viral hepatitis. Liver biopsies from patients diagnosed with chronic viral hepatitis B and C were embedded in paraffin and further used for histological staining and immunohistochemical reactions to detect TGF-beta1 and CTGF. Liver sections stained with trichromic Masson for collagen staining and Gomori's silver impregnation revealed various degrees of liver fibrosis, noted in the METAVIR scale from 1 to 4. Sections with discrete degrees of fibrosis revealed the positivity only in the endothelial cells of liver sinusoids and occasionally in proinflammatory cells from the portal tracts, the number of TGF-beta1-positive cells being directly proportional to the incidence of liver injury. Positive reaction for TGF-beta1 expanded to the cytoplasm of hepatocytes located nearby fibrosis bundles while increasing the parenchymal damage. The expression of CTGF was observed in the classical areas of the hepatic lobule, such as the perisinusoidal spaces around the portal tracts or central veins, but also in the hepatocytes surrounding the fibrotic areas. Regardless of the etiological factor of liver damage, activation of liver cells causes an increased synthesis of TGF-beta1 followed by a CTGF overproduction in various polymorphic hepatic structures, in accordance with the degree of fibrosis.

Corresponding author: Daniela Tache, Assistant Professor, MD, e-mail: danatache@rdslink.ro

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29. The influence of hexestrol diacetate on gametogene function in male rabbit

A. R. Pop, I. Groza, V. Miclaus, Simona Ciupe, Gabriela Mutiu, M. Borzan

The research was performed on 24 male sexually mature domestic rabbits, divided in two equal batches. The rabbits from the first batch were administered 0.5 mg/kg of body weight hexestrol diacetate intramuscularly twice a week for four consecutive weeks. Batch 2 was used for control. The testicular tissue samples obtained after orchidectomy were processed in order to obtain histological samples, stained using the Goldner's trichrome method. Examination of histological sections from the control batch showed that a natural aspect of seminal tubule epithelium, without reported injuries, even discrete. In the experimental batch were recorded a number of changes to all categories of the seminal cells. The severity and extent of damages varied greatly from one seminal tube to another and even from one part to another of the same seminal tube. These changes were the appearance of apoptotic cells and apoptotic bodies, vacuolar degenerescence of spermatocytes and spermatides, syncitialisation of spermatides, areas of necrosis accompanied by severe disruption of the seminiferous epithelium. In some areas, the lesions were so severe that the affected area of the cells forms "basal area". If lesions in the "adluminal area" affects only temporarily gametogenetic function the lesions in the "basal area" are irreversible as there are named "reserve cells" which is the starting point of spermatogenesis. Highlighted issue raised on the opportunity use of hexestrol diacetate in therapy or animal production stimulation as it gametogenetic function in males while their risk transfer to humans through consumption of foods of animal origin.

Corresponding author: Alexandru Raul Pop, Assistant, MD, PhD, e-mail: raulalexandrupop@yahoo.ca

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30. The role of increased plasmatic renin level in the pathogenesis of arterial hypertension in young adults

Manuela Stoicescu, C. Csepento, Gabriela Mutiu, S. Bungau

Introduction: Plasma renin level is an important marker of hypertension in the young adults. The purpose of this study was to determine the role of increased levels of plasmatic renin in the pathogenesis of hypertension in young adults and to highlight the main conditions underlying the pathogenesis of hypertension in young people in these circumstances. Patients and Methods: The group of patients taking part in the study was made of 121 young hypertensive adults (selected from a group of 321 young hypertensive adults), with ages between 18 and 35 years, with elevated blood pressure exceeding 140/90 mmHg in at least three repeated measurements at intervals of one week to exclude white coat phenomenon, or had a blood pressure value greater than 170/100 mmHg at the first measurement and increased plasma renin levels above 4.3 ng/mL. Results and Discussion: Of the 121 young hypertensive patients with increased plasma renin levels, 49 were cases of renal artery stenosis representing 40.50% (p<0.001), eight cases were represented by small unilateral kidneys representing 6.61% (p<0.001), renal cell carcinoma (previously known as "hypernephron" - Grawitz tumor) was responsible for the younger group of patients studied (four cases) representing 3.30% (p<0.001) of the cases of hypertension in the young adults, and 60 cases representing 49.59% were represented by pheochromocytoma. Conclusions: The results show the role of plasma renin dosing as being particularly important in the pathogenesis of secondary hypertension in young adults.

Corresponding author: Manuela Stoicescu, MD, e-mail: manuela_stoicescu@yahoo.com

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31. Study on the histopathological modifications of the dental pulp in occlusal trauma

M. R. Popescu, V. Deva, L. P. Dragomir, Monica Searpe, Mihaela Vatu, A. Stefarta, Anne-Marie Rauten

The diagnosis of occlusion-generated disorders of the dento-maxillary apparatus represents a sensitive stage within the establishment of the therapeutic means for the functional rehabilitation of dental arches. The laborious effort carried out in order to specify the diagnosis resides in the fact that any trauma arising at the level of any component of the stomatognate system may lead to an occlusal dysfunction. The uncured carious processes, besides the pulp and periapical complications, may lead to an occlusal dysfunction through horizontal migrations of teeth resulting in the derangement of the occlusal curvatures as well as through vertical migrations of the teeth opposing a tooth diagnosed with occlusal caries or which largely reduced the coronary height. The dental iatrogenia, besides the eructation anomalies and neuromuscular dysfunction within the oromaxillofacial area also determines the appearance of occlusal dysfunction. The radiological examination through correlation with the clinical manifestations may provide significant data related to the magnitude of the prejudice caused to dento-paradontal units experiencing occlusal trauma. The histopathological study through correlation with the clinical manifestations provides significant data on the tolerance of dento-paradontal units within the occlusal dysfunction. Also, subsequent to the analysis of the possible actions of aggression generated by the occlusal trauma correlations could be determined between the type of the histological lesion of the pulp-dentine complex and the etiopathogenic factors, as well as correlations depending on the damage degree through occlusal trauma of the dental parts involved.

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

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32. Matrix metalloproteinase 9 levels in gingival crevicular fluid in patients after periodontal microsurgery for orthodontic induced gingival hypertrophy

Anne-Marie Rauten, Petra Surlin, B. Oprea, Izabela Silosi, Mihaela Moisa, D. Caramizaru, Mihaela Vatu

Introduction: In this study, we aim to compare the levels of matrix metalloproteinase 9 (MMP9) in the gingival crevicular fluid (GCF), as indicators for healing, in two groups of patients - operated with a classic periodontal surgical technique and the same technique but using a dental microscope. Materials: We included 14 patients with ages between 12 and 26 years, average 14+/-6.2 years. Eight patients were women and six men. All patients presented gingival hypertrophy because of the orthodontic treatment on the mandibular arch. We performed gingivectomy on one-half of the mandibular arch by classic periodontal surgery and on the other half of the mandibular arch by a microscope-assisted gingivectomy. Methods: In the hypertrophied gingiva, the expression of MMP9 was identified using immunohistochemical-staining techniques. For immunological determination of MMP9 in GCF we performed Elisa tests. Results: We found different levels in different moments of the healing process for the two hemiarcades. Conclusions: We consider that faster healing in case of microscope-assisted gingivectomy may be related to the expression of MMP-9 in the GCF.

Corresponding author: Petra Surlin, Associate Professor, PhD, e-mail: vsurlin@hotmail.com

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33. Serum changes induced by intramedullar experimental administration of bisphosphonates

H. Almasan, G. Baciut, Mihaela Baciut, Oana Almasan, S. Bran, L. Oana

Bisphosphonates, stable analogues of pyrophosphate, have the ability to bind to bone molecules, possessing anti-resorbtion properties influenced by the radicals linked to the carbon group of their structure. Bisphosphonates link to the hydroxyapatite of the mineral surfaces and are selectively internalized by osteoclasts, whose activity they inhibit, jeopardizing the osteoblastic activity. The purpose of this study is to determine the influence of intramedular administration (at the hip bone) of bisphosphonates on the serum values of alkaline phosphatase, total Ca, Ca2+, proteins and serum osteocalcin in a lot of experience Wistar rats. Fifteen Wistar rats of experience, five in the control group and 10 in the experimental group. All rats underwent surgery to create a bone defect with a 1.5 mm diameter bone-bur at the right femur transcortical through the medullar canal. Rats from experimental group were divided into two groups: group A, who received Zometa 1 mL single dose intramedular, intraoperative and group B, who received Zometa 1 mL in divided doses daily, 0.1 mL for 10 days. 3 mL of blood from the frontal sinus were collected from each subject at 24 hours, 14 days and 21 days postoperatively. From the blood samples were determined: alkaline phosphatase [U/L], seric proteins [g/dL], total Ca [mmol/L, mg/dL], osteocalcin [mmol/L]. The data were statistically analyzed using the ANOVA test. We found an increase in alkaline phosphatase [U/L] in all groups studied. In group B there was a significant decrease in total Ca levels [mg/dL] throughout the experiment compared with controls (11.82->10.36->9.25 mg/dL; 2.95->2.59->2.31 mmol/L; p=0.001). Ca2+ has changed significantly both in group A (1.18->1.25->1.25 mmol/L; p=0.01) and group B (1.2->1.24->1.13 mmol/L; p=0.02). Serum proteins were significantly reduced both in the control group (9.4->8.5->8.1 g/dL; p=0.03) and the experimental groups A (9.3->8.5->8.3 g/dL; p=0.01) and B (9.9->7.6->7.3 g/dL; p=0.0008). At each stage of bone development, multiple factors act in a coordinated manner that leads to increased local metabolic processes, acting both on the process of bone resorption and bone repair. Healing processes are initiated within 24 hours in both studied groups and the control group; at 14 and 21 days the bone healing processes are compromised directly proportional to the administration manner and dose of bisphosphonates.

Corresponding author: Horea Artimoniu Almasan, PhD student, e-mail: almasanhorea@yahoo.com

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34. Computer-assisted photomicrographic evaluation of root canal morphology after removal of the filling material during retreatment

Luminita Nica, Mihaela Grigorie, D. Rusu, Mirona Mesaros Anghel, Andreea Didilescu, S. I. Stratul

Aim: The purpose of this study was to evaluate the morphology of the apical third of the root canal and the effectiveness of rotary instrumentation for the removal of lateral condensed gutta-percha during endodontic retreatment. Materials and Methods: Thirty roots with canals with regular morphology were prepared to apical size #30 and were filled with gutta-percha/AHPlus using lateral condensation. Digital radiographs of teeth were captured. After two weeks, canals were retreated to size 40 using ProTaper rotary instruments, without solvent. Roots were embedded in resin blocks and reduced incrementally at four different apical levels (at 0.5, 1.0, 1.5 and 2.0 mm from the apical foramen). The sectioned surfaces were observed under a metallographic optical microscope and digital micrographic images were captured and processed. Perimeter covered with root-filling residue was expressed as percentage of total canal perimeter for all specimens. Results: Residue percentage was greater at 2.00 mm from apical level than at other levels. Most residue of filling material in all specimens was observed at 0.5 and at 2.0 mm from apex. The use of ProTaper instruments allowed the removal of gutta-percha and AHPlus sealer in the apical 2 mm in average 50% of the cases. Conclusions: The resulting morphology of the prepared canals allowed root fillings performed with gutta-percha and AHPlus sealer that were efficiently removed by using rotary instruments. More material residue was found adhering to the canal walls in the apical segments of canals. The removal of this residue was enhanced by apical enlargement beyond the diameter of the canal before retreatment.

Corresponding author: Luminita Nica, Senior Lecturer, DDS, PhD, e-mail: nical78@yahoo.com

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35. Investigation of ubiquitin deformation mechanism under induced stretch-compression loads

Mihaela Banu, Simona Patriche, Malina Coman, S. Matsushita, Alice Tofan, A. Epureanu

Proteins folding and unfolding are conformational changes in the life of the proteins, which reveal the modifications in the organization of the molecules inside the chain. It is known that a very organized structure has great amount of entropy and tends to exchange the energy with the environment. The present work presents the results of the atomistic modeling using NAMD software released by the University of Illinois, USA, to reveal the hyper-elasticity under external loads and the progressive conformational shapes that are produced by the H-bonds breaking and recovering. Each H-bond break produces a leap in the internal energy, probably by influencing the bond-bind energy component of the internal energy.

Corresponding author: Mihaela Banu, Professor, PhD, e-mail: mihaela.banu@ugal.ro

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36. Comparative histomorphometric study of bone tissue synthesized after electric and ultrasound stimulation

Malina Coman, Mihaela Hincu, Petra Surlin, Garofita Mateescu, A. Nechita, Mihaela Banu

The clinical use of the alternative therapies in traumatology is conditioned by the knowledge and understanding of their actions on the bone tissue. The hereby study aims at the comparative assessment of the effectiveness of the direct current and ultrasounds in treating the fractures. Thus, we have proceeded to a comparative histological study of the bone tissue in the fractured area and the biomechanical description and the three-dimensional model of the stimulated boneâ??s behavior by using micro-CT X-rays and the finite element analysis. The findings clearly show that the bone, which has been stimulated during a period of two weeks, has regained its functions, that is 85% of the compression one and 95% of the shearing one. These values prove that 90% of the bone structure has healed.

Corresponding author: Malina Coman, Lector, PhD, e-mail: malina.coman@ugal.ro

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37. Psychological status, real pain and antioxidant capacity of plasma, could make the prediction about the post-trauma wound healing

Diana Marcoci, B. Bran, M. Ciurea, H. Parvanescu, Maria Vrabete

Patients submitted to a trauma followed by a surgical intervention, experience pain and discomfort. Exogenous (surgical intervention, anesthesia) postoperative care, and endogenous factors, belonging to patient, especially neuro-psychological disturbances are synergic and participate together, to induce and to amplify the imbalance between oxidants/antioxidants, sustaining the appearance of oxidative injury, and worsening the wound healing. The morphological aspects are not correlated with the pain expression, but the discomfort and agitation have a great influence upon the bad evolution of organism and of wound healing.

Corresponding author: Diana Marcoci, MD, PhD Student, e-mail: dianaomd@yahoo.com

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38. Contributions to the study of morphofunctional interrelations in the liver of the rats treated with certain antipsychotic drugs

Maria Bogdan, Florica Popescu, Fl. Bogdan

The issue of antipsychotics in psychiatry constituted a revolution at the time. The firsts, starting with chlorpromazine represent the conventional antipsychotics, in the last decades there was a new generation of antipsychotics, atypical, which improved the results in treating psychoses. Because, as any drug, it may have adverse effects we aimed an experimental study on rats to observe the toxic potential on liver of both generations of antipsychotics. From the first generation we used chlorpromazine, haloperidol and haloperidol decanoate and from the second, aripiprazole and risperidone. Results of the study show an increased toxicity of chlorpromazine and diminished among the others, without being the same for every drug.

Corresponding author: Maria Bogdan, Assistant Professor, e-mail: bogdan.maria81@yahoo.com

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39. Effects of Alprazolam, Zolpidem and Zopiclone, and of chronic inflammation on peripheral experimental algesia in Wistar rats

Mihaela Zdrinca, Gabriela Mutiu, Maria Bogdan, Luciana Dobjanschi, Angela Antonescu, Ioana Mos, Mariana Muresan, M. Zdrinca, Andreea Antonescu

In the literature, there are some data which indicate that benzodiazepines and other chemical compounds with the same mechanism of action (Diazepam, Chlordiazepoxide, Lorazepam, Zopiclone, etc.) also have other effects. We investigated the effects of experimental chronic inflammation under the administration of some tranquilizers and hypnotics on peripheral algesia induced in rats by "writhing test". Chronic inflammation was induced by "cotton wool granuloma" technique. The "writhing test" consisted in intraperitoneal injection of an irritant agent (acetic acid 0.0025%, 0.4 mL). The animal reacts with a characteristic stretching behavior called writhing. A writhe is indicated by stretching of the abdomen with simultaneous stretching of at least one hind limb. Then, the animals were placed individually into glass beakers and 5 minutes were allowed to elapse. The rats were then observed for a period of 10 minutes and the number of writhes is recorded for each animal. Three drugs were administered by gastric probe: Alprazolam 1 mg/kg, Zolpidem 10 mg/kg and Zopiclone 10 mg/kg. Alprazolam is a triazolobenzodiazepine derivative used as a tranquilizer. Zolpidem is an imidazopyridine with marked sedative-hypnotic effect and it has the same mechanism of action like benzodiazepines. Zopiclone is a cyclopyrrolone with sedative-hypnotic effect used as hypnotic and acts like benzodiazepines. After that, the animals were sacrificed and the weight of cotton wool granuloma was determined. In the same time, the histopatological aspect of granulomatous inflammation was studied. It was found that experimental proliferative inflammation under the action of these drugs was accompanied by a peripheral analgesic activity in "writhing test". The mechanisms of these effects are not fully elucidated. Some explanations are: they act as agonists or antagonists on algesia and inflammation mediators and they have a stimulating effect on peripheral omega3-benzodiazepine receptors ("peripheral-type" receptors).

Corresponding author: Mihaela Zdrinca, Senior Lecturer, MD, PhD, e-mail: mzdrinca@yahoo.com

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40. Multiple organ histopathological changes in broiler chickens fed on genetically modified organism

Daniela Cirnatu, A. Jompan, Anca Ileana Sin, Corina Aurelia Zugravu

Diet can influence the structural characteristics of internal organs. An experiment involving 130 meat broilers was conducted during 42 days (life term for a meat broiler) to study the effect of feed with protein from genetically modified soy. The 1-day-old birds were randomly allocated to five study groups, fed with soy, sunflower, wheat, fish flour, PC starter. In the diet of each group, an amount of protein from soy was replaced with genetically modified soy (I - 0%, II - 25%, III - 50%, IV - 75%, V - 100% protein from genetically modified soy). The level of protein in soy, either modified, or non-modified, was the same. Organs and carcass weights were measured at about 42 days of age of the birds and histopathology exams were performed during May-June 2009. No statistically significant differences were observed in mortality, growth performance variables or carcass and organ yields between broilers consuming diets produced with genetically modified soybean fractions and those consuming diets produced with near-isoline control soybean fractions. Inflammatory and degenerative liver lesions, muscle hypertrophy, hemorrhagic necrosis of bursa, kidney focal tubular necrosis, necrosis and superficial ulceration of bowel and pancreatic dystrophies were found in tissues from broilers fed on protein from genetically modified soy. Different types of lesions found in our study might be due to other causes (parasites, viral) superimposed but their presence exclusively in groups fed with modified soy raises some serious questions about the consequences of use of this type of feed.

Corresponding author: Daniela Cirnatu, Assistant Professor, e-mail: danielacirnatu@yahoo.com

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41. The contribution of laboratory investigations in diagnosis of congenital infections

Simona Neamtu, G. Gaman, Liliana Stanca, Iuliana Buzatu, Lorena Dijmarescu, Magdalena Manolea

Congenital diseases are an important indicator of the degree of development of primary health care, because primary prevention is paramount in diagnosing and diminishing the number of those types of cases. Syphilis is a sexually transmitted chronic infectious disease and with an evolution, often unpredictable. Primary prevention aims to prevent infection of the fetus, while secondary prevention aims for a reduction in the severity of sequels already installed.

Corresponding author: Simona Neamtu, Assistant, MD, PhD, e-mail: simona_0712@yahoo.com

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42. Morphological and immunohistochemical characteristics of a gastric amphicrine tumor: differential diagnosis considerations

Madalina Bosoteanu, C. Bosoteanu, Mariana Deacu, Mariana Aschie

Gastric amphicrine tumors, lesions in which dual epithelial and neuroendocrine differentiation occurs in the same cell, represent neoplasms with a very low incidence. We present the case of a male patient, who suffered a subtotal gastrectomy. Histopathological examination showed a malignant proliferation with glandular, solid and trabecular pattern, composed of round/polygonal, monomorphic cells admixed with scattered signet ring cells. The use of monoclonal antibodies revealed positive immunoreaction of malignant cells for epithelial and neuroendocrine markers. These characteristic features plead for the final diagnosis of amphicrine tumor, based on the co-expression of both epithelial and neuroendocrine markers in the same cells. Differentiation of this entity must be done with collision tumor and with composite tumor, with distinct histopathological features. Rigorous interpretation of dual immunohistochemical expression of neoplastic cells of amphicrine tumor is useful in distinguishing this entity from others with similar morphological characteristics, in order to assure an adequate targeted therapy.

Corresponding author: Madalina Bosoteanu, Lecturer, MD, PhD, e-mail: mbosoteanu@yahoo.com

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43. Endometrial stromal sarcoma developed on outer endometriosis foci

Elena Rosca, Alina Venter, Gabriela Mutiu, Anamaria Dragan, Mihaela Coroi, D. M. Rosca

Introduction: Endometrial stromal tumor is a rare mesenchymal uterine tumor. Endometriosis, an affection of the women during her fertile period, is reported in the pertinent literature to have an incidence varying from 1% to 53%, which shows a wide difference from one study to another. The pathology that grafts onto the endometriosis foci is rare, and the malignant tumoral pathology is extremely rare. The most frequent malignant tumors quoted in the pertinent literature are of the adenocarcinoma type, on the endometriosis, and the alterations of the sarcomatous type can appear extremely rarely. Most sarcomas appear on the foci of the rectosigmoid endometriosis. These rare tumors should not be taken for the mesenchymal gastrointestinal tumors. The recurrence of the malignant alterations of the endometriosis foci is estimated at about 0.7-1% in the pertinent literature. Materials and Methods: We are presenting a case of endometrial stromal sarcoma, developed on foci of outer endometriosis (sigmoid and appendicular endometriosis), diagnosed in our laboratory on two operatory pieces taken from the same woman, at a 2-month interval. The macroscopic and microscopic study was done on operatory pieces, with the routine procedure, macroscopic examination, fixing in 10% formalin, embedding in paraffin and staining with Hematoxylin-Eosin and than we used immunohistochemical markers, for endometrial stroma. Results: The endometrial stromal sarcoma developed on foci of endometriosis is very rare, has an incidence of 6% within the endometrial malignant tumors, with various degrees of malignancy, low malignancy, or high malignancy. Conclusions: We do not exclude the appearance of the endometrial sarcoma on the outer endometriosis foci. Moreover, the endometriosis foci can be considered as pre-malignant conditions

Corresponding author: Elena Rosca, Professor, MD, PhD, e-mail: roscaelena2005@yahoo.com

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44. Retropancreatic cystic lymphangioma - considerations upon a case

V. Surlin, E. Georgescu, Cristiana Dumitrescu, S. Ramboiu, Maria Victoria Comanescu, Mirela Ghilusi

Cystic lymphangiomas are benign tumors developing as an anomaly of the lymphatic channels. Most of them are discovered in children and are located mostly in the head and neck with no differences between sexes. Retroperitoneal locations are rare, clinical symptoms not specific and are discovered incidentally. We report the case of a 32-year-old female sex patient with a cystic tumor situated in the retroperitoneum, behind the pancreas, discovered by abdominal ultrasound during a diagnostic work-out for urinary infection and hypertension. Anamnesis, physical examination and CT-scan could not indicate the nature of the cyst. The cystic tumor had a shape of a butterfly, crossing the midline in front of the aorta and vena cava. A laparotomy was indicated and established the diagnosis of a cystic lymphagioma based on its macroscopic appearance. The tumor was approached from its right side by a generous Kocher maneuver and then from the left by disinsertion of the Treitz ligament and elevating the pancreatic body and splenic vessels. It was completely removed. Postoperative course was uneventful. Histopathologic examination confirmed the diagnosis of benign cystic lymphangioma. At 2 years of follow-up the patient was free of recidive. Conclusions: Retropancreatic cystic lymphangioma are rarely seen in adults. Precise diagnosis is made at laparotomy, imagistic test give details about walls, content, relationship with the surrounding structures and major blood vessels. Complete surgical resection represents the treatment of choice avoiding recurrence.

Corresponding author: Valeriu Surlin, Lecturer, MD, PhD, e-mail: vsurlin@gmail.com

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45. Urothelial carcinoma of the renal pelvis associated with cystic disease of the kidney

Mariana Deacu, Madalina Bosoteanu, Mariana Aschie, Gabriela Baltatescu, V. Sarbu, Mariana Bardas

Primary tumors of the renal pelvis and ureter account for about 8% of all urinary tract tumors. More than 90% of them are urothelial carcinomas. On the other hand, unilateral multicystic renal disease is an uncommon pathologic condition that may be mistaken for unilateral autosomal dominant polycystic kidney disease, multilocular cystic nephroma or cystic neoplasm. We present the case of a 54-year-old male known with arterial hypertension, admitted in the Second Surgery Department of Emergency County Hospital, Constanta, with intense right flank and right lumbar pain. This symptom started one month before hospital admission. Based on clinical features and imaging evaluations we established a presumptive diagnosis of unilateral autosomal dominant polycystic kidney disease. For these reasons, total right nephrectomy was performed. Pathologic examination of the nephrectomy specimen revealed high-grade urothelial carcinoma of the renal pelvis associated with unilateral multicystic renal disease. The particularity of this case lies in the uncommon association between two rare renal pathological conditions diagnosed by pathological examination.

Corresponding author: Mariana Deacu, Lecturer, MD, PhD, e-mail: deacu_mariana@yahoo.com

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46. Parietal abdominal endometriosis following Cesarean section

M. Pasalega, C. Mirea, I. D. Vilcea, I. Vasile, I. E. Plesea, F. Calota, C. Mesina, H. Parvanescu, M. Baluta

Endometriosis is a pathological feature induced by the presence and ectopic development of islets of endometrial active cells. The most common site of occurrence is the genital system, causing specific gynecological pathology. The extragenital localization of endometriosis is rare, but it is more severe and it may have a malignant local evolution, although its structures remain benign. The endometrial inclusions in the abdominal wall scar are iatrogenic "implants", created at the same time with the surgical operation, performed on patients with genital endometriosis. The only curable treatment of this topography of endometriosis is the surgical removal of all the pathological tissue, through a large excision. The hormonal therapy is adjuvant. Our study presents three cases treated in our clinic; the most important objective was to establish the etiological diagnosis and, subsequently, the large excision of the lesions.

Corresponding author: Marin Pasalega, MD, e-mail: pasmar@mail.com

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47. Considerations upon a case of synchronous primary malignancies: adenocarcinoma of the sigmoid and clear cell carcinoma of the right kidney

A. Kozokic, V. Surlin, B. Petrovic, V. Petrovic, G. Prvanovic, I. Beraru, Simona Cheregi

Synchronous primary colon and renal cancer is a rare but real clinical entity reported with variable incidence. An 81-year-old man admitted for abdominal pain and melena is diagnosed with right colic tumor by colonoscopy and with simultaneous right kidney tumor by CT-scan. The patient is adequately prepared and scheduled for laparotomy. Both tumors are resected in the same surgical session with curative intent - right hemicolectomy and right radical nephrectomy. The patient recovered well and was discharged after 10 postoperative days. The microscopic examination indicated an adenocarcinoma in the sigmoid and a clear cell renal carcinoma in the kidney. Immunohistochemical staining did not find any compatibility between those tumors. The patient started chemotherapy and is under appropriate oncologic follow-up. Modern investigations allow detection of simultaneous malignancies making possible the planned simultaneous resection of both. Histopathologic examination will proof the diagnosis of primary different malignancies.

Corresponding author: Valeriu Surlin, Lecturer, MD, PhD, e-mail: vsurlin@gmail.com

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48. Squamous carcinoma of the conjunctiva

Mihaela Cristiana Coroi, Elena Rosca, Gabriela Mutiu, T. Coroi

We present the case of a 63-year-old patient who was hospitalized for pain and loss of vision in the right eye (RE). Clinical examination reveals a conjunctival tumor of 1.5/1 cm infiltrating the cornea and sclera. Biopsy and histopathological examination showed the existence of a spinocellular carcinoma.

Corresponding author: Mihaela Cristiana Coroi, Associate Professor, MD, PhD, e-mail: opticlar@gmail.com

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49. Clinical and histological correlations in alveolar bone osteosynthesis using biological materials of cow origin

M. Suciu, C. Kelemen, O. S. Cotoi, Felicia Toma

Periodontitis is a set of inflammatory diseases affecting the periodontium, i.e., the tissues that surround and support the teeth. Periodontitis involves progressive loss of the alveolar bone around the teeth, and if left untreated, can lead to the loosening and subsequent loss of teeth and is one of the most common diseases worldwide. Modern techniques of treatment consist of guided bone regeneration, in cases of massive bone loss. We present a case of a middle age male with no risk factors, suffering from infected chronic marginal periodontitis with III/IV degree of mobility at the lower right canine and III degree of mobility at the lower incisors. X-ray exam reveals massive bone resorption in the anterior part of the mandible, especially in the right canine area. Because the buccal and lingual cortical bone were missing in the canine area, it was impossible for the bone to heal after the extraction of 4.3., without bone augmentation. The histopathological exam revealed the aspect of an organic protein matrix, partially calcified and the presence of some isolated cells with osteocytes like morphology. The difficulties of this procedure are to position and to close the flap without tension in order to facilitate the healing and bone cells formation - osteoblasts and osteocytes. The bone augmentation using biological material proved to be successful in treating massive bone defects in order to insert dental implants.

Corresponding author: Mircea Suciu, Associate Professor, MD, PhD, e-mail: oralmed2000@yahoo.com

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50. In memoriam Professor Bogdan Zaharia, MD, PhD (1941-2010)

I. E. Plesea

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Corresponding author: -

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