K-ras expression--marker of dysplasia and cancer.

Molecular and epidemiological data indicated that the presence of HPV virus is not sufficient to induce transformation, suggesting the implication of other several cellular factors. Constitutive activation of the Ras signaling pathway is an important component of malignant progression for a number of different cancers. In this context, the objectives of our study were: the quantitative assessment of the K-ras gene expression changes in the development of the HPV positive cervical cancers. We observed that the K-ras mRNA expression levels did not gradually increase with the severity of injury. The mRNA expression in the ASCUS increased 2.02 times as compared with the control group, while in LSIL group only 1.76 times. However ras expression was increased in the HSIL/cancer group by 2.27 times when was reported to the control group. The presence of low risk HPV infection (IrHPV) does not lead to increased ras expression, remaining at baseline, but K-ras expression was increased in the presence of high risk HPV infection (hrHPV). In addition, we noted that in hrHPV single infections ras expression is increased (0.96 +/- 0.48) comparing with hrHPV co-infections. Our findings indicate that high expression of ras among hrHPV infection can be a marker of cervical cancer development.

P16INK4A--A possible marker in HPV persistence screening.

UNLABELLED: The aim of this study was to investigate the correlation of p161NK4a expression levels with the cytological group of cervical carcinogenesis (NILM, ASCUS, LSIL, HSIL, cancer groups), in order to establish its value as potential diagnostic marker. METHODS: The smears obtained from 50 women with/without suggestive HPV infection pathology were subjected to cytological investigations. The viral testing was based on the detection of HPV DNA using the INNOLIPA kit, while the semiquantitative expression levels of p16INK4a were estimated by RT-PCR. RESULTS: p16INK4a expression level was correlated with the cytological degree of cervical lesions. In LSIL patients, p16INK4a values were 1.36 times greater than in NILM subjects (p = 0.07). In HSIL/cancer patients, p16INK4a values were 2.38 times greater than in NILM patients (p = 0.002). We also noticed significant differences between ASCUS: HSIL group (p = 0.02) and LSIL: HSIL (p = 0.07) group. The p16INK4a expression level was dependent of HPV genotype, p16INK4a mRNA presence being correlated with the presence of hrHPV in low and high risk lesions.

Electrosurgical loop excision/conisation for cervical intraepithelial neoplasia in an algorithm that excludes punch biopsy--a study of 210 cases.

UNLABELLED: This study aims to evaluate the use of LLETZ/conisation in an algorithm that excludes the colposcopically guided biopsy. MATERIAL AND METHODS: The study was carried out on 210 patients with LLETZ/conisation, performed in our service in 2 years. They were selected by pap smear, colposcopy, HPV genotyping, without punch biopsy. RESULTS: The pathological results on the excision specimen showed: benign lesion 10%, CIN 1/condyloma 58%, CIN 2 18%, CIN 3/CIS 11%, microinvasion 2% and invasion 1%. The Pap test showed: HGSIL 27%, LGSIL 56%, ASCUS 13%, and normal/benign in 4%. The therapeutic efficiency of the excisional treatment showed that there was a 9.5% excessive treatment, 14.8 residual lesions, 3 cases of hemorrhage, 2 cervical stenosis, and 7 cases with specimen alteration that made the pathological diagnostic difficult or impossible. In conclusion, the LLETZ/conisation are ambulatory procedures with an acceptable rate of over-treatment and residual lesions, and reduced rate of complication.

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

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.

Immunocytochemical expression of p16INK4a and HPV L1 capsid proteins as predictive markers of the cervical lesions progression risk.

UNLABELLED: Genital HPV infections are extremely common but most of them are spontaneously cleared by the host immune response. The main problem is how to identify the HPV-HR positive patients who are at risk of progressive disease. AIM: The purpose of this study was to investigate the uterine cervix lesions concerning the HPV status appreciated through the immunocytochemical expression of the L1 HPV and p16INK4a proteins. MATERIAL AND METHODS: 76 women who tested positive for HPV were selected from a cohort of 374 patients. In this study were detected the immunochemical expression of HPV L1 capsid protein and p16INK4a in LBC samples. RESULTS: The p16INK4a positive rate was expressed in 56.57% of all the cases. The percentage grew from 0% in NILM cases to 100% in SCCs cases (p-value <0.00001). The HPV L1 capsid protein positive was expressed in 12.50% of NILM cases, 33.33% of ASC-US, 50% of LSIL, 18.51% of HSILs cases, but 0% in the SCC group (p-value = 0.01). The L1-/p16+ pattern was found in 21.87% of LSIL, 81.48% of HSIL, and 100% of SCC cases (p-value <0.00001). The association of these two markers (L1 and p16INK4a) raises the accuracy of the diagnostic from 64% for HPV L1 capsid protein and respectively 87% for p16INK4a to 91% when they are associated. CONCLUSIONS: The combination of L1 capsid protein and p16 appears to be useful for an early diagnosis and may be able to identify the patients with risk of lesion progression.

[Epidemiological data on shock in forensic medicine]. / Date epidemiologice asupra socului in cazuistica medico-legala.

UNLABELLED: The shock is a relative frequent diagnostic in forensic medicine. MATERIAL AND METHOD: We realized a statistical study, using 323 deaths through diverse shock typology, registered within the Forensic Medicine Institute of Iasi in 2000-2009. The concerned variables were: the spread of cases on years, the clinical forms, age, sex, provenience environment, volemic feature, the survival time from the shock to the death, the spread of complications on the form of the shock and survival, the juridical way of death. Our data were introduced in the EXCEL database and statisticaly analyzed by SPSS program. RESULTS: We obtained statistic data on shock typology, based on the survival time and based on the complications of shock types. CONCLUSION: Relevant statistic data were obtained on the relations between the factors that generated the shock, the complications induced and the shock types, related also to the juridical way of death.

[Evaluation of p16INK4a and Ki-67 proteins expression in cervical intraepithelial neoplasia and their correlation with HPV-HR infection]. / Evaluarea expresiei proteinelor p16nk4a si ki-67 în neoplazia intraepiteliala cervicala si corelarea acestora cu infectia HPV-HR.

OBJECTIVE: The aim of our study was to evaluate the expression of p16INK4A and Ki-67 in cervical intraepithelial neoplasia (CIN) and to correlate the high-risk human papillomavirus (HPV) infection with these two biomarkers. MATERIAL AND METHOD: Sixty patients were included in our study. All of the patient underwent colposcopy-guided biopsy to asses the grade of CIN. For the immunohistochemistry exam we used p161NK4A and Ki-67 antibody and their expression was semiquantitatively classified in 4 classes: 0, 1, 2 and 3. HPV genotyping was used to detect high-risk HPV. RESULTS: 7 of our cases were benign (11.66%); 18 cases presented CIN1 (30%), 14 cases CIN2 (23.33%), 16 cases CIN3 (26.66%) and 5 cases invasive squamous cell carcinoma (8.33%). We observed that the expression of p16INK4A and Ki-67 were positively associated with CIN grade, p161NK4A expressions increased significantly with high-risk HPV infection. We observed a positive correlation between the expression of the p16INK4A and Ki-67. CONCLUSION: In our study, the expression of p16 and Ki-67 were positively related to the CIN grade. p16INK4A expressions of high-risk HPV specimens significantly increased more than Ki-67. Therefore, in the diagnosis of CIN and high-risk HPV infection, p16INK4A can be a useful biomarker.

[Endometrial hyperplasia predicts a better prognosis in endometrial carcinoma]. / Hiperplazia endometriala--factor de prognostic favorabil în carcinomul endometrial.

AIM: To evaluate the significance of the associated Endometrial Hyperplasia in Endometrial Carcinoma. STUDY DESIGN: 93 hysterectomy specimens for endometrial carcinoma were histologically reviewed on paraffin sections stained H&E and tricromic VG. RESULTS: 45 cases were associated with endometrial hyperplasia; 48 cases without endometrial hyperplasia were: 35 cases with the entire endometrial cavity occupied by carcinoma, 11 associated with endometrial atrophy and 2 with normal proliferative endometrium. In this second group, 37 cases (77%) were in advanced stages. A correlation was found between the presence of Endometrial Hyperplasia and the degree of differentiation, the FIGO stage of the carcinoma, and the age of the patient. CONCLUSIONS: Endometrial Carcinomas associated with Endometrial Hyperplasia involve younger women and have lower aggressivity than those without hyperplasia. Endometrial Hyperplasia may demonstrate a favorable prognosis in Endometrial Carcinomas.

[Morphological correlations between endometrial hyperplasias, uterine leiomyoma and ovarian associated lesions]. / Corelatii morfologice între leziunile hiperplazice endometriale, leiomiomul uterin si leziunile ovariene asociate.

Morphological evaluation and correlation of endometrial hyperplasias, leiomyoma and ovarian estrogen producing lesions. 390 specimens of hysterectomy associated or not with ovarectomy were processed by routine technique and sections stained by H&E staining. 316 cases presented different degrees of endometrial hyperplasias associated with leiomyoma. In 117 from 170 cases the ovary presented estrogen secreting lesions (follicular cyst, polycystic ovary, stromal hyperplasia, granulosa cell tumours). The highest frequency of the uterine and ovarian lesions was in the decades 41-50 years, and 31-40 years. Leiomyoma and endometrial hyperplasia, develop in a hormonal context, most frequently characterized by micropolicystic ovaries. The risk is higher in the perimenopausal period. The most frequent type is simple hyperplasia suggesting a rare progression to highest grades and a possible protective role of leiomyoma as target tissue which capture estrogens.

[Value of steroid receptors assessment in atypical endometrial hyperplasia]. / Valoarea determinarii receptorilor hormonilor steroizi în hiperplazia endometriala atipica.

To evaluate if the immunohistochemical assessment of steroid receptors is helpful in distinguishing Atypical Hyperplasia from Well Differentiated Endometrial Carcinoma, the morphological and immunohistochemical features for steroid receptors were studied in 46 cases of atypical hyperplasia alone or associated with other degrees of hyperplasia or adenocarcinoma. The estrogen and progesterone receptors were determined by the avidin-biotin complex technique using monoclonal antibodies on paraffin embedded tissue. The diagnosis of atypical hyperplasia was based on the observation of epithelial atypias and the absence of the stromal invasion. The positive reaction for steroid receptors was moderate, very variable from case to case, heterogeneous and quite similar to that observed in carcinoma. The morphological identification of the stromal invasion is the most reliable criterion in differentiating endometrial carcinoma from atypical hyperplasia. The steroid receptors assessment is not valuable in this purpose but may be useful in cases in which a hormonal therapy is recommended.

[The prognostic molecular factors in precancerous lesions and nonmelanotic skin carcinoma]. / Factori moleculari în leziunile precanceroase si carcinomul cutanat.

The most common cutaneous carcinoma, which includes basal cell carcinoma (BCC) and squamous cell carcinomas (SCC), accounts for approximately 90% or more of all skin malignancies. Actinic keratosis, bowenoid type of actinic keratosis, Bowen's disease or SCC in situ have been considered precursors of skin SCC. Authors present the histopathological criteria of precursors of skin SCC, variants invasive SCC and BCC. Also, one discusses about the roles of immunohistochemical staining of gene products in evidence of molecular changes in cell that might occur according to the transformation from precancerous lesions to SCC.

[Medical and social implication of cervical cancer screening]. / Implicatiile medico-sociale ale screening-ului cancerului de col uterin.

Invasive cervical cancer represents a major health problem in Romania. When there is a functioning screening network, the incidence of invasive lesions has a marked reduction, this program being the most successful cancer prevention program of all times. 80% of the actual incidence and mortality from this disease occurs in countries without an organized screening. A good infrastructure of the cytopathologist--practitioner relation must be achieved in order to reach the standard of early detection of the preinvasive cervical lesions.

[Malignant mixed mesodermal tumor: histo- and cytopathologic correlations]. / Tumora mixta mezodermica maligna. Corelatii cito-histopatologice.

To correlate the cytopathological and the histopathological findings in uterine mixed mesodermal malignant tumor (MMMT) we have examined the cervical smear, endometrial curettage and hysterectomy specimen of a patient diagnosed with uterine tumor. The smear was stained by Papanicolaou staining and the tissue processed by routine technique and stained H&E. The original cytological diagnosis was adenosquamous carcinoma. The histopathological diagnosis was MMMT of heterologous type. A review of the smear revealed features which may orientate the diagnosis: multinucleate cells, isolated cells with cyanophilic cytoplasm, hyperchromatic nuclei and prominent nucleoli, elongated cyanophilic cells of sarcomatous origin. We conclude that the cytopathological diagnosis of the MMMT in cervical smears is very difficult. This may be sustained by the evidence of more cell types and cellular features orientating to a sarcomatous origin. The most important differential diagnosis is adenosquamous carcinoma.

[Endometrial carcinoma in situ. Morphological and immunohistochemical aspects]. / Carcinom endometrial in situ. Aspecte morfologice si imunohistochemice.

A possible precursor for invasive endometrial carcinoma is an in situ stage, but this notion is very much debated. We present a case of endometrial carcinoma diagnosed in a patient with post-menopausal bleeding. The morphological examination of the hysterectomy specimen, processed by routine technique, put in evidence focal atypias of the glandular epithelium in a group of few endometrial glands, on a background of atrophy without a desmoplastic stromal reaction. The immunohistochemical investigation of the steroid receptors and p53 protein, using monoclonal antibodies in the ABC technique, revealed the p53 positivity and the absence of the receptors in the focal malignant areas. The surface epithelium had the same immunohistochemical aspect. These aspects were decisive in precising that the malignant change was primarily in the surface epithelium with secondary extension in some glands, the final diagnosis being in situ serous papillary carcinoma.

Histological study of Hodgkin disease (HD), an immunohistochemical identification of the cells surrounding Sternberg-Reed cells.

In this study, we analyze histological types of 77 cases H.D. and examine immunohistochemically the reactive cells surrounding Reed-Sternberg cells (40 cases). On the current techniques sections of lymph nodes were def ned H.D. types after the new criteria (Bennett & col., 1985, Buttler, 1992). Immunologic marker studies were performed on paraffin sections of H.D. tissue utilizing the avidin--biotin peroxidase complex technique with limited panel of monoclonal antibodies OKT3, OKB1, OKM1. On histopathologic grounds we established the diagnosis of lymphocytic predominance type in 6 cases--8%, nodular sclerosis type in 36 cases--47%, from which first degree--24 cases and second degree--12 cases, mixed cellularity type in 28 cases--36% and lymphocytic depletion type in 7 cases--9%. Immunohistochemically, in our 40 H.D. cases, the number of T. lymphocytes in all H.D. types, was highly variable, ranging from a minority to a vast majority of cells directly surrounding the Reed-Sternberg cells. These lymphocytes were positive for OKT3, which were more in tissue involved by nodular sclerosis or mixed cellularity types than in the lymphocytic predominance type. Thus, in our few cases of lymphocytic predominance type, the reactive proliferation was consisted of B. cells along side with a few T cells. Also the OKM1 positive mononuclear cells were few. In conclusion, our data support the necessity to extend of several immunohistochemical investigations used for distinguishing H.D. disease from benign hyperplasia and from non-lymphoid malignancies, and for concerning the functional lymphocytic composition and the role of monocyte-macrophage system in the immunological alteration in H.D.

[Anal squamous cell carcinoma synchronous with rectal adenocarcinoma]. / Carcinom scuamocelular anal sincron cu adenocarcinom de rect prezentare de caz.

Most often colorectal carcinoma occurs single; synchronous multiple carcinomas usually develop at widely disparate sites. We report the case of a 75-year-old male, accusing rectal bleeding, disturbances in bowel transit and weight loss. The rectoscopy examination revealed a fungating, bleeding tumor located 5 cm from anal verge. Pathological diagnostic of the endo-biopsy was ulcerated moderate differentiated adenocarcinoma. Patient underwent surgical amputation of the rectum with lymphadenectomy. Microscopical examination of the surgical specimens confirmed the presence of the adenocarcinoma adjacent to a squamous cell carcinoma, moderate differentiated, with reduced keratinization, infiltrative. Also, 2 from the 7 lymph nodes presented squamous cell carcinoma metastases. The most important differential diagnostic is a rectal adenosquamous carcinoma. Prognostic depends on stage of the disease, generally being worse than of the corresponding adenocarcinoma, and can be improved by radio- and chemotherapy.

[ASCUS of metaplastic type. Cyto-histopathological correlations]. / ASCUS de tip metaplazic. Corelatii cito-histopatologice.

We evaluated the cytopathological changes and the prognostic significance of atypical squamous metaplastic cells in cervical smears. 50 ASCUS cases were divided in two groups: 25 cases in different settings and 25 of metaplastic type. All cases were re-evaluated after 6 months and when necessary, verified by biopsy. The second cytological diagnosis was: group I--13 normal, 2 LSIL, 40 ASCUS; group II--6 normal, 2 LSIL, 2 HSIL and 15 ASCUS. The HPV was positive in 5 cases in group I and in 9 cases in group II. In ASCUS persistent cases the biopsy revealed: in group I--3 CIN 1 and 1 CIN 2 cases; in group II--1 CIN 1, 2 CIN II, 2 CIN III and one case of microinvasive carcinoma. In conclusion, atypical metaplastic cells are more frequently involved in serious cervical lesions.

[An immunohistochemical study of the steroid hormone receptors in endometrial hyperplasia]. / Studiul imunohistochimic al receptorilor hormonilor steroizi în hiperplaziile endometriale.

AIM: Evaluation of the estrogen and progesterone receptors (ER and PR) in different degrees of endometrial hyperplasias. METHODS: 10 cases of each degree of endometrial hyperplasia (simple, complex and atypical) were analyzed using the avidin-biotin-peroxidase technique and monoclonal antibodies applied to formalin-fixed, paraffin-embedded tissue. RESULTS: We found similar, high level of both ER and PR in simple and complex hyperplasias and a significant decrease of these in atypical hyperplasia. CONCLUSIONS: Endometrial hyperplasias develop in a setting of estrogen excess. This explains the high level of endometrial receptors and the response to progesterone in typical hyperplasia. In atypical hyperplasia, the decrease of steroid receptors results in a low sensibility of this lesion to progestative therapy, but there are cases with high receptor levels which could have a chance for hormonal therapy.

[p53 and steroid receptors support double endometrial carcinogenesis]. / p53 si receptorii steroizi sustin o dubla carcinogeneza endometriala.

AIM: This study investigated the p53 positivity correlated with the receptor status, in different grades and stages of endometrial carcinoma. METHODS: The study included 88 endometrioid-type carcinomas and 5 non-endometrioid-type carcinomas. Paraffin sections were used for the ordinary histological diagnosis and for immunohistochemical diagnosis by avidin-biotin-peroxidase technique. RESULTS: p53 positivity was identified in 10 endometrioid-type carcinomas (11.4%) and 3 non-endometrioid type carcinomas (60%). Most of the p53 positive cases were receptor negative and observed in advanced stages and histological grades. No associated endometrial hyperplasia was p53 positive, while a positive intraepithelial non-endometrioid carcinoma was identified. CONCLUSIONS: p53 positivity is most frequently identified in non-endometrioid type, receptor negative adenocarcinomas, even in a precursor lesion as it is intraepithelial carcinoma, while in endometrioid, receptor positive carcinomas it appear in late stages of development, never being identified in precursor lesions.

[Diagnosis of gastric cancer on endobiopsies using the WHO classification]. / Aplicarea clasificarii OMS în evaluarea carcinomului gastric diagnosticat pe endobiopsie.

Gastric cancer (GC), one of the most frequent malignancies can be early detected on endobiopsies. Our aim was to evaluate histologically the GC on endobiopsies, using WHO 2000 and Lauren classifications. The study included 2424 gastric endobiopsies, routine processed; sections stained with HE, Giemsa, PAS and AB. GC was diagnosed in 451 cases (19%), mostly in men (311 cases--69%). The highest incidence was in 60-69 year-old aged patients (34%). Using Lauren classification, 279 cases were included in diffuse type (62%), 167 intestinal-type (37%) and 5 (1%) mixed type. Each of these three types were also histologically analyzed considering the WHO classification. We pointed out that GC can be diagnosed histologically on endobiopsy specimens, although it is difficult to diagnose the GC--mixt type, due to reduced size of endobiopsies. Lauren classification (including the two major types: diffuse and intestinal) is very useful, especially if correlated with histological criteria of WHO classification.