Literature data on peculiarities of melanocytic skin neoplasms in children and adolescents have been analyzed in order to evaluate topicality of research into the diagnostic markers complex for skin melanoma in these patients as well as, if the conclusion is in the affirmative, to characterize the would-be complex of such markers. Skin melanoma in children and adolescents is a heterogeneous tumor group which when compared with adult skin melanoma possesses both some features of similarity and a number of essential distinctions. Differential diagnostics between pigment nevus and melanoma remains rather intricate for professionals. These difficulties might be minimized by application of molecular markers. As a possible approach to differential diagnostics between nevi and malignant skin melanoma in children it seems worthwhile to try the following steps: to reveal INK4a/ARF locus lesions and to detect the protein p16^INK4a by immunohistochemistry; to check for amplifications in 11q13 chromosomal region as well as to carry out immunohistochemical detection of cyclin D1 (protein CCND1) in cellular nuclei; to reveal HLA class I antigens on surfaces of pigment cell neoplasms in immunohistochemical test.

Target delivery of antitumor drugs to cancer cells seems to be the very promising way of cancer therapy. The study on the application of immunoliposomes as nanocontainers for anticancer drugs started in the 90-ies. Immunoliposomal drug formulations of antitumor preparations have some advantages over traditional forms of drugs: lipid capsule reduces toxicity of drug due to the selective delivery to tumor and improves its bioavailability. However, despite these benefits, at present immunoliposomal drugs application is limited in the clinic. This review discusses current research status in field of development immunoliposomes and the possible targets for anticancer immuno-liposomes.

Background. Malignant phenotype of cancer cells and metastatic potency of the tumor are determined by genetic factors. In addition, normal biological environment, including the nano-vesicles or exosomes, plays an important role in regulation of the structural and functional characteristics of malignant cells. Objective: presented study was aimed to evaluate mechanisms and to estimate effect of interaction of plasma exosomes and breast cancer cells in experimental conditions. Materials and methods. We used breast cancer cell culture MDA-MB-231 and exosomes isolated from plasma and cultural medium. Exosomes were analyzed by dynamic light scattering method and western blotting. Functional effects of exosomes were evaluated in in vitro and in vivo models. Results. In the present study we demonstrated that plasma exosomes stimulate the adhesion and the motility of breast cancer cells and induce the process of metastatic dissemination. Contact interaction of exosomes with cell surface is sufficient for stimulatory effect that is mediated by exosomal fibronectin and FAK-dependent signaling cascade. Conclusions. Further investigation of plasma exosomes structure and functions is required to better understand their input in regulation of malignant cell phenotype. This research has a potential to provide novel approaches for cancer therapy.

Background. Class III beta-tubulin (TUBB3) is one of the eight beta tubulin isotypes identified in human. It is constitutively expressed in brain and testis but not in lung tissue. TUBB3 is also known to appear in solid tumors, in particular in non-small cell lung cancer (NSCLC), and it is often associated with poor prognosis and resistance to taxanes and Vinka alkaloids. Objective: We have suggested that TUBB3 expression may cover not only the primary tumor but also a wide adjacent area of morphologically normal lung tissue. To check the hypothesis we have measured TUBB3 expression both in the non-small cell lung carcinoma (NSCLC) and remote lung tissue derived from the same lung. 60surgical biopsy specimens of NSCLC and morphologically normal lung tissue of 30patients were investigated. Materials and methods. Biopsy specimens were converted to suspension, fixed in 4 % formaldehyde and analyzed by flow cytometry using monoclonal antibodies to TUBB3. The method was developed in the laboratory of medicinal chemistry research Institute of experimental diagnostics and therapy of tumors (N.N. Blokhin Russian Cancer Research Center) and patented. Results. Fraction of TUBB3-positive cells in the group of adjacent lung tissue specimens was lower compared to NSCLC group but still positive in the majority of adjacent lung tissue specimens (25 of 30) and achieved up to 39 % of cells. Conclusion. We suggest that TUBB3 expression in adjacent morphologically normal lung tissue indicates presence of transformed and potentially malignant cells far from the primary site.

Objective: the purpose of the study is research of the changes in phenotypic and genetic characteristics of melanoma cells Mel Ibr cultured in growth medium with low concentration (5 %) of embryo calf serum. Materials and methods. Cell line Меl Ibr was pre-cultured in RPMI-1640 medium with 5 % FBS for 3passages, and then 200-400 cells were replaced on the Petri dish with a diameter of 60 mm and left in the incubator for 14 days to form colonies. Colonies of spindle form cells were cultivated in the same conditions for more than 30 passages. Cells were stained according to the method of Leishman to investigate the morphology. The expression of antigens was determined in the reaction of indirect immunofluorescence. Results. Exposure of Mel Ibr human melanoma cells to low concentration of embryo calf serum resulted in the appearance of a subclone with morphological, immunological and cytogenetical characteristics differed to those of parental cells. The subclone cells distinguished with spindle form, more small size, large nucleus which broadcasted on whole cytoplasm, formation spheroid and sharply reduced percentage of HLA-DR (+) and CD54 (+) cells, a significantly elevated percentage of CD63 (+) cells, and appearance of CD133 (+). The subclone karyotype differed to those of parental cells. Conclusoin. The subclone cells were characterized by the same features as stem tumor cells and could be use as tumor progression model.

Objective: Evaluation of antitumor activity of a novel alkylnitrosoureas derivative Ormustine (an alkylnitrosocarbamoyl L-ornithine) in mouse lymphoid leukemia models. Materials and methods Antitumor activity of Ormustine has been evaluated in B6D2F1 mice with ascites form of leukemia (L1210, L1210/arenosa, L1210/citrullin and P388) and the solid (P388) form. In this study we used preparations from the alkylnitrosourea group: Ormustine, Aranoza and Lizomustine. Treatment of animals was started 24 hours after inoculation of leukemia intraperitoneally, and 48 hours after inoculation subcutaneous P388. Drugs in a wide range of doses were administrated once intravenously. The follow up period of the animals continued until their death. Criteria of antitumor effect were increasing life expectancy and cure. Evaluation criteria of antitumor effect was the increase in life of experimental mice compared to control ones. Results. Antitumor activity of a novel alkylnitrosoureas derivative, Ormustine has been studied in vivo on the growth of transplanted lymphoid leukemia, such as L1210 (ascites version) and P388 (ascites and solid tumor). Effective dose of single intravenous injection Ormustine against lymphoid leukemia L1210 and P388 was 125 mg/kg. The drug effectively inhibited growth of experimental leukemia. The significant part of the mice with limfoleukemia has been cured. We have also established the single intravenous therapeutic dose of Ormustine on L1210 and Р388 leukemia - 125mg/kg of body weight. Conclusion. The data obtained characterizes Ormustin as a promissing anticancer drug.

Background. Cancer of the esophagus is the sixth most common malignancy worldwide and there is a remarkable variation in the incidence of esophageal cancer in different parts of the world. Objective: The present study was designed to evaluate chemopreventive activity of lycopene on esophagus carcinogenesis induced by N-methyl-N-benzylnitrosourea in rats. Methods. The study was conducted on male Wistar rats with body weight at the beginning 160-180g. Tumors of esophagus were induced by N-methyl-N-benzylnitrosourea in dose of 0.5 mg/kg body weight in 10 % ethanol given daily with drinking water during 8 weeks. Results. Lycopene given in dose of 50 mg/kg reduced incidence of tumors of esophagus from 100 % in N-methyl-N-benzylnitrosourea group to 44 % (р < 0.05). Lycopene also inhibited tumor multiplicity, the standard end point in this tumor model 2,4 times in comparison with N-methyl-N-benzylnitrosourea group (p < 0.005). The average score of neoplastic changes decreased 2.4 times in lycopene group (р < 0.001) and indicates inhibition of malignization of neoplastic changes in target organ. Additionalfeeding of N-methyl-N-benzylni- trosourea-treated rats with lycopene preventedformation of the imbalance in lipid peroxidation - antioxidant defense system by prevention of inhibition of antioxidant enzymes and accumulation of malonic dialdehyde. Conclusion. At this stage of our study, we can conclude that lycopene appears to be worthy of consideration as an antioxidant, and antitumor medicine.

Background. Present study was dedicated to identification and evaluation of the biologically active components in multiphytoadapto-gene preparation phytomix-40 (PhM-40). It consists of forty plant extracts components including Panax ginseng. PhM-40 demonstrates wide spectrum of biological effects including antimutagenic, antitumor, radioprotective, hormone-modulating, antioxidant, neuropro-tective and ummunomodulating activities. Objective: of this study was to identify and to quantify ginsenosides content in the preparation. Materials and method. Experimental design included the comparison of ginsenosides content evaluation in PhM-40, comparison preparation (which contains similar to PhM-40 plant extracts components but another rations) and Panax ginseng extract by means of LC-MS/MS spectroscopy method. Two different gradients were used: the short and the long ones. By means of the short one rapid compound identification with economical consumption of chemicals was possible as well as the long one allowed us to identify compounds with better resolution capability. Commercially available ginsenosides were the standards for calibration. Literature data were also used for ginsenosides identification. Results. High through liquid chromatography method with tandem mass-spectrometry was successfully used for ginsenosides identification quantitatively and qualitatively in plant preparation PhM-40. The data obtained show the ginsenosides qualitative composition which has been identified as Rb, Rb₂, Rc, Rd, Rg_p Rg₂, Re, Rf, Ro, malonyl-Rb_p -Rb₂/Rb₃/Rc, -Rd enumeration in PhM-40

The aim of the study was to compare the biodistribution and the photodynamic efficacy of the chlorin е₆ trimethyl ester (3MeChl) and its galactose-substituted in pyrrole A derivative (3MeChl_gal_NoAc). Materials and methods. In vitro experiments were carried out on cultivated murine Lewis lung carcinoma (LLC) cells using multipara-metric mode. Biodistribution and therapeutic efficacy of 3MeChl 3MeChl_gal_NoAc were studied in BDF1 mice bearing s. c. transplanted LLC. The biodistribution study was performed using local fluorescence spectroscopy. For evaluation of the photoinduced antitu- mor activity the photosensitizer was administered i. v. on day 7 of tumor growth in doses varyingfrom 0.5 to 7.5 mg/kg. The tumors were irradiated by LED source with a wavelength of the emitted light 661+/ - 16 nm, at various drug-light time interval (from 5 to 120 min), and at energy density 90 J/cm². Results. It was shown in cultured murine Lewis lung carcinoma cells that 3MeChl_gal_NoAc has higher photo-induced cytotoxicity than the unsubstituted compound (IC₀₀ value of 25 ± 1.5 nM and 171 ± 4.0 nM, respectively). When administered intravenously to mice with subcutaneous LLC a normalized fluorescence intensity of 3MeChl in tumor reached the maximum value (18.4 ± 0.7 a. u.) within 60 min, and for MeChlgalNoAc it reached the maximum value (34.1 ± 6.9 a. u.) within 15 min. A higher fluorescent contrast between tumor and normal skin or muscle was registered for 3MeChl (up to 7.4 and 4.1, respectively) than for 3MeChl_gal_NoAc (up to 5.0 and 2.5, respectively). Having similar biodistribution profile in normal organs and tissues of tumor-bearing mice, 3MeChl_gal_ NoAc was eliminatedfrom the body faster than 3MeChl. Intravenous administration of 3MeChl_gal_NoAc in dose of 5.0 mg/kg following 15 min light exposure (light dose 90 J/cm²) with a drug-light interval of 5 min caused a high antitumor effect, 100 % of animals were cured. However, a rapid elimination of the galactosyl derivative from the tumor tissue, a low selectivity of photodynamic damage of the tumor, as well as a narrow range of highly effective therapeutic doses considerably limit prospects for this modification of chlorin е₆ for photodynamic treatment of the malignant tumors.

Objective. Aim of this work was to create a stable liposomal dosage form of native hydrophobic antitumor compound from the group of indolocarbazoles - LHS-1208. Materials and methods. Quantitative analysis of the drug content in liposomes was determined by spectrophotometry with a standard sample at X = 320 ± 2 nm. The encapsulation was investigated as the ratio of LHS-1208 concentration in the liposomal dispersion after extrusion through nylon membrane filters 0.22 ^m “Pall” to concentration LHS-1208 in liposomal dispersions before filtration. pH of the liposome was determitaned by the method of potentiometry. The size of liposomes was evaluated by nanosizer. Cytotoxic activity was studied by MTT-test. Results. Experimental liposomal models of LHS-1208 with different molar ratios of the components were obtained and analyzed. Composition with the molar ratios LHS-1208: lecithin 1:150, and lecithin : cholesterol: PEG-2000 - 1:0,2:0,003 was selected. Encapsulation percentage of LHS-1208 was 94 % and size of the vesicles was 185±10 nm. Cytotoxic activity of liposomal LHS-1208 was studied, IC₀ was 1.24 ^g/ml. Conclusion. As a result of complex pharmaceutical research determined the optimum composition of the components and the technology for production of liposomal dosage form LHS-1208.

Background. Urticaria is а common disease characterized by the formation on the skin itchy blisters. It is easy to diagnose an urticaria, but it isn’t always simple to treat because of various severity, frequent resistance to antihistamine drugs, a substantial reduction in quality of life. Objective. The purpose of the study is to carry out the retrospective analysis of medical case history of patients with acute and chronic urticaria for identification of the most significant etiological factors of the disease for the optimization of diagnostics and treatment of such patients. Materials and methods. There were analysed 307 medical cards of the patients with various forms of urticaria who were on treatment in the allergology department from 2008 till 2014. The diagnosis was established on the basis on the results of the clinical methods conforming to the standards of diagnosis of this disease, and also specific allergological research. Results. Though patients with acute and chronic urticaria pass examination and treatment in the hospital according to the one medical economic standard, patients with a chronic urticaria demand wider range of diagnostic actions. Respectively, their medical examination is more expensive. Conclusion. Despite on a wide range of the possible reasons of an urticaria, there are patients at whom the provocative factor isn»t taped. Thus, the urticaria is a multicausal disease at which complex examination of the patient is required.

Introduction. In the formation of recurrent and/or prolonged chronic inflammatory diseases of the upper respiratory tract in addition to the known, previously studied, factors that play an important role in the violation of the human immune system at the level of local (mucosal) and systemic immunity and activation of herpes virus group (Epstein Virus Barr virus, cytomegalovirus, herpes simplex virus type 6). In the treatment of herpesvirus infections, activation of antiviral immunity in domestic medicine used drugs interferon inducer. Objective. To evaluate the efficacy of interferon inductor Amiksin® in patients with chronic recurrent inflammatory diseases of the upper respiratory tract. Materials and methods. Based on separation of “Allergy and Immunotherapy” SSC “Institute of Immunology” FMBA studied low-molecular interferon inductor Amixin® (JSC “Pharmstandard-Tomskhimpharm”, Russia) in the treatment of chronic recurrent inflammatory diseases of the upper respiratory tract. The study included 40 patients between men and women, aged 18 to 65years old with a history of recurrent chronic inflammatory diseases of the upper respiratory tract. Clinical research methods included a medical history, previous efficiency of the treatment, the presence of comorbidities. Laboratory methods include bacteriological crop on flora in the material from the oropharynx and the detection of DNA viruses of herpes group in saliva. Patients of the main group, after clinical and laboratory examination, prescribed therapy with Amixin®. Patients in both groups received symptomatic therapy. The total duration of observation of each patient was 3 months. Results. Amiksin® receiving the drug in patients with acute exacerbation of chronic recurrent upper respiratory tract inflammatory diseases contributed to a more rapid relief of general and local symptoms. Also it found that reduces performance Amiksin® average viral load against Epstein-Barr virus. Over the next 3 months follow-up, 25 % of patients the main group marked exacerbation study pathology of the upper respiratory tract, in the control group of patients with recurrent exacerbations were more - 60 %, indicating that preventive action Amiksin® therapy. Conclusions: The use of low-molecular interferon inductor Amiksin®(JSC “Pharmstandard-Tomskhimpharm”, Russia) in the combined therapy showed good efficacy in reducing the concentration of chronic viral infections in the oropharynx and prevention of chronic relapsing inflammatory diseases of the upper respiratory tract.

Background. Identification of cellular proteins can be performed by indirect immunofluorescence assay using flow cytometry. However, this method allows to detect cellular proteins that are either on the cell surface or inside the cell and cannot demonstrate a protein distribution within cellular compartments, particularly in nucleus. Meanwhile, the nuclear localization of the protein of interest in many respects gives an indication of the mechanisms of its action. Therefore, the development of the protocol of extraction of nuclei suitable for analysis on a flow cytometer is able to complement the information about the localization and functional significance of many nucleus-associated proteins. Objective. The aim of this study was to develope a method for extraction, purification and stabilization of intact nuclei suitable for flow cytofluorimetry analysis. Materials and methods. In this work we studied the nuclear localization of the receptor type 1 for vascular endothelial growth factor (VEGF-R1). The A431 human cancer cell line was used as an object of the study. The quality of extracted nuclei was assessed by microscopic examination of stained smears of nuclear suspension. Expression of the receptor was determined by indirect immunofluorescence assay using flow cytometry. Results. New method was successfully applied to obtain the suspension of intact cellular nuclei that is crucial to perform further flow cytometry. Applied method revealed the presence of the receptor type 1 for vascular endothelial growth factor at the external nuclear membrane and inside of the nucleus. Interesting to note that the presence of the receptor type 1 for vascular endothelial growth factor inside ofthe nucleus was 3,8 times as much as its surface location (17,9 ± 1,04 % and 4,8 + 0,26 % respectively). Conclusions. The new method of extraction, purification and stabilization ofthe nuclei is applicable for proteins identification by flow cytometry. In combination with other methods (ICC, Western blotting, etc.) flow cytometry of intact nuclei is able to complement the information about the properties of nucleus-associated proteins.