TUMOR INFILTRATING LYMPHOCYTES PECULIARITIES IN DIFFERENT HISTOPATHOLOGICAL AND MOLECULAR SUBTYPES OF GASTRIC CARCINOMA.

Gastric carcinoma represents one of the major causes of cancer related mortality worldwide. Recently, immunotherapeutic means gave the new promise for the treatment of gastric carcinoma, although, not all patients benefit from this type of treatment. Tumor infiltrating lymphocytes (TILs) are considered as one of the promising prognostic and predictive biomarkers in solid tumors. However, the presence of TILs is not well characterised in different types of gastric cancer. The aim of our study was to characterise TILs profile in different histopathological and molecular subtypes of gastric carcinomas. We used standard haematoxylin and eosin staining (H&E) for evaluation of TILs and immunohistochemistry to detect molecular markers, including CDH1, Ki67, p53 and Her2. The results of our study revealed that TILs status varies significantly in different histological and molecular subtypes of gastric adenocarcinoma, which might be the reason for different prognosis in these patients. Also, high TILs status is significantly related to the presence of p53 mutations in both enteric type and diffuse type gastric carcinomas, which can be further explored for immunotherapeutic options in these patients.

THE STUDY OF THE EXPRESSION OF CDH1, KI67, P53 AND HER2 IN DIFFUSE GASTRIC CARCINOMA.

Gastric cancer represents the fifth most common malignancy and third most common cause of cancer deaths worldwide. There are two major types of gastric carcinoma: enteric type and diffuse type. Diffuse type gastric cancer is frequently associated with the mutations in E-cadherin coding gene CDH1. In cases of CDH1 mutations complete gastrectomy is indicated. CDH1 mutations are reflected with CDH1 protein loss by immunohistochemistry. The relationship between CDH1 mutations and other markers of tumour aggressiveness, such as tumour proliferation index and the presence of p53 mutations and Her2 amplification is not well studied. Therefore, the aim of our study was to analyse the correlation between CDH1 loss and the expression of Ki67 proliferation marker, mutant oncoprotein p53 and Her2. Standard immunohistochemistry was used to detect the following antigens: CDH1 (MCH-38, Invitrogen), Ki67 (EP5, Bio SB), p53 (DO-7, Leica) and Her2 (EP3, Bio SB). The study results showed that CDH1 mutations, reflected with CDH1 protein loss by immunohistochemistry are detected in 40% of diffuse gastric carcinomas, whilst it is not detected in enteric type gastric carcinomas. Diffuse gastric carcinomas with CDH1 mutations are characterised with more aggressive phenotype, particularly with the presence of higher Ki67 labelling index, p53 mutations and the presence of Her2 positivity. In cases of histological diagnosis of diffuse gastric carcinoma CDH1 testing is recommended.

[State of the hypothalamo-hypophyseal-ovarian system in young nullipara women with ectopia of the cervix of uterus during taking the hormonal contraceptives].

Background diseases of the cervix of the uterus play one of the leading roles in the structure of gynecological pathology and present the risk of the development precancerous changes. Ectopia is observed in the structure of precancerous processes of the cervix of the uterus in 38, 8% of women and in 42, 2% cases of gynecological diseases. Our aim is to investigate the content of gonadotropic and steroid hormones in the blood plasma of young nullipara women with different types of ectopia during taking hormonal contraceptives. Cohort study has been carried out by using simple blind method. The quantitative data analyses were performed using the Statistical Package for the Social Sciences (SPSS) in order to reveal the correlation between taking of oral hormonal contraceptives and the hormone content in the blood plasma among young nullipara women with different types of ectopia. Descriptive statistics were calculated for all the study variables. The results displayed correlation between taking the oral hormonal contraceptives and changes of hormonal background in young women with ectopia of the cervix of the uterus during taking hormonal contraceptives. The study show that the secretions of gonadotropic hormone and ovary hormone peculiarities depend on the type of ectopia of the cervix of the uterus. The effect of hormonal contraception on cervix of the uterus of young nullipara women with ectopia was investigated. The oral contraceptive, Exluton is recommended in young nullipara women with ectopia.

[Hormonal contraception and cervix of the uterus pathology].

The effect of hormonal contraception on cervix of the uterus of young nullipara women with ectopia was investigated. Cohort study was carried out by using simple blind method. By means of statistical data manipulation correlation between the application of hormonal contraceptives and changes of colposcopy pictures in the zone of pathology was displayed. Excluton showed low percent anomalous colpscopy picture in pathology zone. Excluton was declared as medicine of choice for nullipara women with ectopia. Still it is necessary to conduct a strong dynamic control of cervix of the uterus (colposcopy and pap-smear test) in order to avoid complications.

[Ectopia of cervix of the uterus and hormonal contraception].

The aim of the study was the investigation of impact of different hormonal contraceptive drugs on cervix of uterus of young nullipara women with ectopia. Cytologic smears were examined using Pappanikolau method. Cohort study was carried out by using simple blind method. The data were treated by statistics packet (SPSS). The results displayed correlation between taking the hormonal contraceptives Rigevidon and Marvelon and in transformation of ectopia into micro glandular hyperplasia, which did not occur in taking medicine Exluton. Drug Exluton is recommended for young nullipara women with ectopia to exclude micro glandular hyperplasia. In case of prescribing monophase contraceptives for young nullipara women with ectopia cytological control of endo- and exo-cervix is recommended.

[The immunohistochemical characteristics of cervical glandular intraepithelial neoplasias].

The aim of our study was to identify immunohistochemical diagnostic criteria for cervical glandular intraepithelial neoplasias (CGIN). We examined 136 women with cytological diagnosis of atypical endocervical cells. These patients were divided into three groups based on a grade of the lesion: 35 patients with CGIN1 (group I), 72 patients with CGIN2 (group II), 28 patients with CGIN3 (group III). Endocervical curettages were examined by hematoxylin-eosin and immunohistochemistry using monoclonal antibodies against Ki-67, EpAg, MNF116, CEA, EMA. We used histological algorithm created by us. The comparative analysis of immunohistochemical results showed that expression of Ki-67 is seen in CGIN 2 and significantly increased in CGIN3 (p<0,05) which indicates increased proliferative activity of glandular cells in relation to increased grade of lesion. The differences in the expression of MNF116 and EMA are not statistically significant (p>0,05) which indicates that the expression of these epithelium specific markers does not change according to the grade of atypia and carcinogenesis (they can be used for determination of tumor phenotype). The expression of CEA and EpAg is strongly increased in CGIN2 and CGIN3 (p<0,05) indicating their potential role in carcinogenesis. The results suggest that evaluation of a grade of cervical glandular intraepithelial neoplasia should be based on histological and immunohistochemical studies. The morphometric algorithm should include the following criteria: type of lining epithelium (cubical, columnar), nuclear cytoplasmic index (<1, >1, =1), stratification, hyper- and hypochromasia, size and amount of nucleoli, and stromal-parenchymal ratio. The immunohistochemical study should include the expression of proliferation marker (Ki-67), carcinoembryonic antigen (CEA)and Epithelial Antigen (EpAg). We recommend the classification of CGIN into two types: low grade cervical glandular intraepithelial neoplasia (CGIN 1) and high grade cervical glandular intraepithelial neoplasia including CGIN 2 and CGIN 3.

[Histological and morphological characteristics of the different stages of cervical glandular intraepithelial neoplasia].

Currently there is no world system classification of glandular hyperplasia of uterine cervix (endocervix). Different authors give nonmonosemantic estimations to each type of cervical hyperplasia. Cervical Glandular Intraepithelial Neoplasia (CGIN) or Glandular Dysplasia is considered as a precancerous lesion of endocervix. Morphometric investigations have shown that for estimation of the degree of the endocervical glandular and intraepithelial neoplasia it is necessary the quantitative evaluation of the following criteria: type of epithelium--prismal, columnar, estimation of nucl/cytopl index >1, <1, =1, stratification, hyperchromatosis, hypochromatosis, big nucleoli, small nucleoli, stromal/parenchimal index >1, <1, =1. Based on our observations we suppose that glandular neoplasia must be divided into low grade intraepithelial neoplasia -CGIN 1, and high grade intraepithelial neoplasia -CGIN 2, and CGIN 3.

[Oxidative processes in the cervical and vaginal epithelial tissue in patients with HPV infection].

Genital human papilloma virus (HPV) infection is the sexually transmitted disease. HPV induces the proliferation of epithelial cells. In the areas of viral inclusions vulvovaginal warts, papillomas and condylomas are developing. Some serotypes of HPV are known as a significant risk factor of cervical cancer. The aim of our study was to detect the conditions of oxidation processes in vaginal and cervical tissues in patients with HPV infection. According to our results, in patients with HPV, intensification of production of reactive forms of nitrogen and oxygen in vaginal and cervical tissues takes place which is the result of activation of leukocytes, other phagocyte cells and chaotic flow of blood in damaged tissue, as well as interchanging periods of hypoxia and reperfusion.

[Determination of cytomorphological criteria for differential diagnostics of variable stages of endocervical glandular cells atypia].

The aim of the investigation was to determine the criteria of differential diagnostics of different stages atypia of endocervical cells in women of different age groups. For ECCIN3, out of 12 diagnostic criteria, the most important and statistically significant were the following five: a) high N/C index > 5,54; b) number of mitoses > 10,3; c) number of produced rosettes >3,9; d) rough redistribution of chromatin >4,64; e) hyperchromatosis >1,87. In case of ECCIN3 the majority of patients (82,75%) had infection of the cervix. In 13,79% of patients HPV infection has been documented. Hemorrhagic discharge was a very characteristic symptom (57,12%) of ECCN3.