The participation of PDCD4 protein in molecular mechanisms of key importance in carcinogenesis makes the assessment of the level of its synthesis in tissues important for assessing the early diagnosis of malignant tumors. The aim of the presented study was to develop a method for evaluation of the results of immunohistochemical staining of the PDCD4 protein. Material and methods. The material for the study included 49 samples of human gastric adenocarcinoma tissue and 80 cases of endoscopic biopsy examination of the gastric mucosa of patients with dyspeptic complaints. Morphological changes in biopsies corresponded to chronic gastritis with varying degrees of inflammation. Rabbit monoclonal anti-PDCD4 antibodies (clone EP 102, abcam, USA) and a detection system (Diagnostic BioSystems, USA) were used to detect PDCD4 protein by immunohistochemical method. Results. The developed can be used for semi-quantitative visual assessment of synthesis of regulatory protein PDCD4 in histological sections of tissue by immunohistochemical method with detection in nucleus and cytoplasma. That is ensured by calculating an expression index of PDCD4 in tissues when summing up the nuclei expression and cytoplasm expression. Nuclear expression represents a product of staining frequency of nuclei (0–3 points) and semi-quantitative visual assessment of intensity of coloring nuclei (0–3 points). Cytoplasmic expression represents semi-quantitative visual assessment of intensity of cytoplasm (0–3 points). Conclusion. The proposed method makes it possible to evaluate the synthesis of the PDCD4 regulatory protein in tissues with greater sensitivity and specificity, which makes it possible to detect a violation of PDCD4 expression in epithelial cells at the early stages of tumor cell transformation.

Aim: to evaluate the possibility of the MMR-system status, microsatellite instability (MSI) usage in the differential diagnosis of gastric mucosa dysplasia, determination of the gastric adenocarcinoma development risk. Material and methods. The study included gastric mucosa specimens of 75 patients: 25 with high-grade dysplasia, 25 with low-grade dysplasia, 25 were indefinite for dysplasia. Gastrobiopsy specimens were examined histologically, immunohistochemically using mouse monoclonal antibodies (Diagnostic BioSystems, USA) to the MMR system proteins: MLH-1 (clone G168-15, dilution 1:50), MSH2 (clone DBM15.82, dilution 1:100), MSH6 (clone 44, dilution 1:50), PMS2 (clone A16-4, ready to use). MSI was studied with multiplex PCR evaluation of DNA microsatellites (NR-21, NR-24, NR-27, BAT-25, BAT-26) from paraffin sections, their analysis with capillary electrophoresis. The obtained data were processed with the Statistica 10.0 (StatSoft, USA), presented using descriptive, analytical statistics. VOSviewer (1.6.20) was used to visualize the bibliometric analysis. Results. MMR-deficient cases were found in low (2.8 %) and high-grade (2.8 %) dysplasia with the immunohistochemical evaluation of MMR-system proteins in gastric mucosa specimens. In all indefinite for dysplasia cases MMR-system proteins remained unaffected. Three MSI-positive cases (6.5 %) were detected by PCR with two low-grade dysplasia, one high-grade dysplasia cases. All identified cases were also immunohistochemically MSI-positive. Conclusion. Determination of MSI can be used as an auxiliary study within a panel of biomarkers aimed to support the decision-making of a pathologist in the alternative of “indefinite for dysplasia” or “definite dysplasia — obligate precancer”.

Introduction Diagnosis of the gastric mucosa atrophy represents an important problem, the solution of which depends on the possibility of secondary prevention of gastric cancer. A possible way of solution is the use of immunohistochemical markers - proteins associated with cellular remodeling of gastric mucosa, PDCD4 and CDX2.The aim of the work is to evaluate the possibility of using immunohistochemical markers PDCD4 and CDX-2 to diagnose atrophy of the gastric mucosa in chronic gastritis and increase the informative value of biopsy examination.Materials and method The object of the study was 155 cases of biopsy examination of the gastric mucosa of patients with chronic gastritis (5 fragments per case − 775 biopsy specimens). A comparative semi-quantitative assessment of immunohistochemical expression of CDX2, PDCD4 at different stages of chronic gastritis was performed. Spearman correlation coefficient was used to assess correlation relationship.Results There were no statistically significant differences in the level of PDCD4 in studied samples depending on the stage of chronic gastritis, p=0.06. Statistically significant increase of CDX2sum index in progressing stage of chronic atrophic gastritis (p=0.005), demonstrated a pronounced positive correlation r=0.70 (p<0.01).Discussion According to the results obtained, it is shown that the decline in PDCD4 protein does not occur with the progression of atrophy severity. Complementary use of immunohistochemical marker CDX2 is able to give an idea of the presence and severity of both metaplastic and absolute atrophic changes in the gastric mucosa.Conclusion Equally high level of PDCD4 protein index in the gastric mucosa at different stages of chronic gastritis excludes the possibility of its use as an immunohistochemical marker of atrophy. Semi-quantitative immunohistochemical index of CDX2 protein can be used as an additional marker in decision support system for assessment of atrophic changes in gastric mucosa.

The article presents the comparative results of the morphological studies of the mucous membrane of the tympanic cavity in experimental exudative and purulent otitis media (EOM, POM). Thirty male Wister rats with a body weight between 180 g and 250 g were included into the experiment and divided into four groups, including intact individuals with simulated EOM and POM, ultrasonic regional lymphotropic therapy in EOM modeling. The material for morphological examination taken on the 4th and 12th days was processed according to the standard method and studied by light microscopy at different magnifications (×40, ×100, ×200, ×400). It was shown that in the initial stage (4th day) of the process in the second and third groups a similar reaction takes place in the form of the formation of serous exudate and leukocyte infiltration of the mucous membrane of the tympanic cavity. By the 12th day of the experiment, in each otitis forms, a discrepancy in morphological manifestations was detected, such as sluggish serous inflammation, edema, round cell leukocyte infiltration of the mucoperiost, without pronounced destruction of the epithelial tissue in exudative otitis media; and prominent destruction of the epithelium with disintegration of its deep structures in purulent otitis. Ultrasound regional lymphotropic therapy of exudative otitis media led to a decrease in the severity of the infiltrative reaction and restoration of the integrity of the epithelial layer of the mucous membrane of the tympanic cavity. The conducted studies incline the authors to recognize the inflammatory theory of the pathogenesis of exudative otitis media.

The aim of the study was to evaluate the potential of miR-21 expression in the gastric mucosa as the marker of early stage gastric adenocarcinoma and cancer development. Material and methods . The study included the following materials: the first group consisted of surgical material of 60 stomachs (gastric cancer of intestinal type), in each case, 1 fragment of tumor tissue and 4 specimens of the gastric mucosa of the distant zone (2 – antrum mucosa and 2 – fundal mucosa) were taken; the second group consisted of biopsy material of 62 cases in patients with chronic gastritis (5 specimens according to the OLGA-system protocol). The level of miR-21 expression in tissues was determined by reverse transcription polymerase chain reaction. Results. miR-21 expression in samples with gastric cancer was significantly higher (median – 158, p=0.0006) compared with the group of endoscopic biopsies of the gastric mucosa (median – 42). The highest level of expression was observed in adenocarcinoma tissues (median – 270), while there were no significant differences when comparing it with the level of miR-21 expression in samples of the distant zone mucosa (p=0.2).тIncreased miR-21 expression may indicate the high risk group of developing gastric adenocarcinoma, probablyтbefore the appearance of histological signs of neoplasia. Conclusion. Regression analysis performed to confirm the predictive potential of miR-21 expression demonstrated 13-fold increased risk of adenocarcinoma development associated with an increase in miR-21 levels for every 50 units of relative expression normalized to small U6 RNA. The constructed statistical model, which includes morphological criteria for evaluating the gastric mucosa and miR-21 expression, may reflect the degree of violation of cellular epigenetic molecular pathways and allows increasing the prognostic value of a biopsy investigation and the accuracy of a stratified assessment of the gastric adenocarcinoma risk development

Objective: to identify the of bisoprolol effectiveness predictors in patients with stable angina after myocardial infarction.Materials and methods: 107 patients with stable angina who underwent myocardial infarction aged 35–65 years (mean age 54,7 ± 6,2 years) were examined in an open, comparative, register study, of which groups were generated: with the achieved and unreached target heart rate (60 or less in 1 minute). Additionally, the respondents were stratified into subsamples by the presence of ADRB1 gene Arg389Gly polymorphism. All patients underwent a 5-minute study of heart rate variability (HRV) in the background sample and in active orthostasis and the determination of the ADRB1 gene polymorphism (Arg389Gly, rs1801253) by PCR. All patients received bisoprolol in the selected optimal maximum-tolerated dose.The main results: respondents who did not reach the target heart rate significantly more often complained about heartbeat and heart rhythm interruptions (p = 0,003), increased incidence of cardiac pain episodes (p = 0,02), noted a high demand for nitrates (p = 0,03) and significantly more often sought medical help. In respondents with the achieved target heart rate, sympathetic influences were less expressed in the background HRV sample with significantly higher parasympathetic influences expression by contrast. The number of sympathicotonics in terms of HRV at rest was significantly higher among respondents with unreached heart rate. When comparing the frequencies of alleles in respondents with the achieved target heart rate, a significant (p = 0.0001) prevalence of carriers of the Gly allele of the ADRB1 gene Arg389Gly polymorphism was revealed. The heart rate in carriers of the Gly allele in homo-or heterozygous form was significantly lower than in carriers of the homozygous genotype Arg389Arg.Conclusion: The predictors of the bisoprolol effectiveness in achieving the target heart rate in patients with stable angina after MI are: a stress index less than 104.5 cu. in the background sample of a 5-minute HRV, the presence of the polymorphic Gly allele of the ADRB1 gene Arg389Gly polymorphism, the age of less than 46 years, and the total spectrum power in the background HRV sample of more than 1309 ms.

The aim of the study was to assess expression of microRNAs (miRNAs) influencing the key carcinogenesis pathways in the tissues of gastric mucosa in patients with H. pylori-associated chronic gastritis and gastric cancer of intestinal type (directly in the tumor tissue and mucosa of the stomach with morphological signs of chronic gastritis). Materials and Methods. Objects of investigations were fragments of gastric mucosa of the antral part of the stomach with H. pylori-associated chronic gastritis: 10 were bioptates with minimal changes, 9 bioplates with chronic gastritis, and specimens of surgical material taken from the stomachs with gastric cancer of intestinal type: 28 were tumor fragments and 22 the tissue from the sites beyond tumor growth (chronic atrophic gastritis). The number of miRNAs miR-146., miR-155, miR-21, miR-223, miR-192, miR-200b, miR-221, miR-375 in paraffin sections was determined by a real-time semi-quantitative hairpin RT-PCR assay. Results. The analysis of miR-21 and miR-223 showed the most statistically significant results among the investigated miRNAs. The expression level of oncogenic miR-21 and miR-223 significantly and successively increases in the range of the examined diseases from H. pylori-associated chronic gastritis to intestinal type gastric adenocarcinoma. Conclusion. MiRNAs miR-21 and miR-223 may be considered as perspective biomarkers for assessing the risk of development, early diagnosis, and prognosis of intestinal type gastric cancer.

Carriage of polymorphic alleles of genes of cytokines-interleukines IL-1β, IL-1RN, TNFα, IL-4 can be a specific feature of chronic periodontitis patients. Genetic tests can be used to predict the course of the disease at its early manifestations. Objective: To establish the relationship of clinical manifestations of periodontal disease, inflammatory cytokines gene polymorphism and systemic levels of cytokine production. Periodontal tissue assessment and cone-beam computed tomography (CBCT) were performed in 150 periodontitis patients. A molecular--genetic testing for the presence of polymorphic alleles of genes IL-1β -511 C>T and +3953 C>T, IL-1RN (VNTR intron 2), IL-4 (VNTR intron 3), TNFα-308 G>A; content determined IL-1β, TNFα, IL-4 in peripheral blood was carried out in 150 patients with periodontitis and 150 healthy donors. Based on the analysis of the speed and nature of the supporting bone resorption and clinical manifestations patients are divided in "aggressive", "moderately progressive" and "slowly progressive" periodontits course groups. Disease severity was associated with distribution of genotypes and alleles of polymorphic genes cytokine IL-1RN (VNTR intron 2), TNFα-308 G>A and IL-4 (VNTR intron 3); haplotype IL-1β-511 TIL-1β +3953 T/IL-1RN 2R. There was no statistically significant difference in systemic level of IL-1β, TNFα and IL-4 between periodontitis groups but the donor level of cytokines was 2-4 times less.

The aim of this study was to determine the morphological changes of human gastric mucosa infected with Helicobacter pylori (HP) depending on polymorphism of the genes coding both interleukin-1beta and interleukin-1 receptor antagonist. 106 juveniles with HP-induced chronic gastritis were studied along with 44 juveniles with gastritis, not associated with HP. In patients with HP-associated gastritis, the significant increase in epithelial proliferation and apoptosis indexes was observed. Epithelium proliferation zone was expanded in the direction of surface foveolar epithelium and into the depth of glands; pronounced lympho-plasma cell infiltration of mucosal lamina propria was observed. Cytokine gene polymorphisms were evaluated by PCR. The results suggest an association between more pronounced inflammation of gastric mucosa with the T allele of interleukin-1beta C -511 C > T gene polymorphous locus (Chi2 = 14.006; p = 0.001). No association was found of the interleukin-1beta C +3953 T gene and the variable number of tandem repeats in the 2nd introne of interleukin 1 receptor antagonist gene with morphological changes in the gastric mucosa.

The purpose of this work was to study morphological features of gastric mucosa depending on polymorphism of IL-l1beta and IL-1RN genes in children with chronic gastritis infected with different strains of Helicobacter pylori 0. We examined 106 children with HP-associated gastritis. The results suggest association of mucosal inflammation with T allele in polymorphous C-511T locus of IL-1beta (beta2 = 14,006; alpha = 0.001). agA + strains of HP were more frequently identified in patients with erosive ulcerative defects of gastric mucosa than in children with superficial gastritis (p < 0.05). The possibility of gene typing to form a group of patients with the unfavourable post-eradication clinical course for the prevention of duodenal ulcer and stomach cancer is discussed.