Vol 16, No 3 (2021)

Regulatory T lymphocytes (Treg) control the activity of immune cells and suppress the development of inflammation, maintaining the immune balance necessary for the body. Dysfunctions of Tregs are associated with the pathogenesis of autoimmune and oncological diseases. With systemic and organ-specific autoimmune reactions, as well as organ transplantation, a decrease in the function of Tregs is observed. While in the course of oncogenesis, the activity of Tregs prevents the development of an adequate immune response to tumor antigens, promotes the processes of angiogenesis and uncontrolled growth of transformed cells. Taking into account the important function of Tregs in the control of autoimmunity and oncogenesis, approaches to immunotherapy of inflammatory pathologies based on autologous and donor Tregs, as well as methods of activating an antitumor immune response as a result of selective blockade of the functional activity of Tregs, are being actively developed. The review provides an overview of technologies for modulating the activity of Tregs for the treatment of cancer, autoimmunity and adverse reactions after transplantation.

Development of reproductive medicine, namely, extracorporal fertilization, set the task of preservation and preserving of biological suitability of female and men's gametes for scientific community. Opening of a method of a vitrification showed its benefit before other methods of a cryopreservation and became the encouraging event in development of cryobank of gametes. In this work the history of emergence of a method of a cryopreservation is described; modern aspects of a cryopreservation of gametes; shortcomings and benefits of various techniques of selection of full-fledged gametes are reflected; some protocols of carrying out a cryo-preservation are provided; short characteristic of separately used cryoprotectors and their various combinations capable more effectively to protect the freezing objects from damage, than, each of cryoprotectors separately is given. It is noted that in modern literature special attention is paid to consideration of mechanisms of cryodamages and cryoprotection during the freezing and a vitrification. Characteristics of the changes happening in gametes after the procedure of freezing and thawing are discussed. Modern techniques of assessment of viability of gametes before and after a cryopreservation are given. It is shown that violation of compaction of chromatin and fragmentation of DNA in morphologically not changed spermiya has negative impact and on quality of embryos. The benefit of a structurally functional condition of oocytes of mammals at a cryopreservation of oocytes in structure is emphasized an oocyte-kumulyusnykh of complexes in comparison with the oocytes which ripened in the kultivatsionny environment.

Neurofibromatosis type 1 (NF1) is an autosomal dominant hereditary tumor syndrome that occurs on average with a frequency of 1: 3000 people. Clinical features of NF1 include cafe-au-lait macules on the skin, multiple cutaneous, subcutaneous, and plexi-form neurofibromas, Lisch nodules, and optic gliomas. The disease is also characterized by damage to the musculoskeletal system, impaired intelligence and behavior. The cause of the disease is germinal mutations in the NF1 gene, which encodes the oncosuppressor neurofibromin. NF1 is characterized by pronounced polymorphism of clinical manifestations, from erased to severe, without geno-phenotypic correlations. Therefore, the role of modifier genes in the pathogenesis of NF1 is assumed. We carried out a clinical-epidemiological and molecular-genetic study of patients with NF1 from the Republic of Bashkortostan (RB). We searched for intragenic mutations by sequencing 57 exons of the NF1 gene, and identified deletions of the entire gene using microsatellite analysis. The prevalence of NF1 in RB is 10 per 100 000. We identified 14 intragenic mutations in the NF1 gene in 20 patients with NF1 and 2 extended deletion of NF1 gene in 3 patients from 2 unrelated families. We did not find a correlation between the type of mutation and the characteristics of the clinical manifestations of the disease. To determine the possible influence of modifier genes on pathogenesis of NF1, we carried out a comparative analysis of the clinical manifestations of NF1 depending on inheritance, ethnicity and association of systemic manifestations. Among the clinical manifestations of NF1 in RB, there was a low frequency of optic nerve gliomas (5,25%) and plexiform neurofibromas (5%). The incidence of brain cysts was 4,25% among patients with NF1. We identified 9 mutations for the first time in the world, 5 out of 14 identified mutations are known (c.2806A>T, c.2991-1G>C, c.3158C>G, c.4537C>T, c.6792C>A). The distribution of mutation types turned out to be random. We identified a protective role of crossbreeding for the development of severe manifestations of NF1. We determined a high incidence of scoliosis, short stature, facial dysmorphism and chest deformity in the inheritance of NF1 from the mother, which suggests the influence of modifier genes on the pathogenesis of NF1.

In the past few decades, there has been a decline in male fertility worldwide. The article is devoted to the study of the reproductive health of subfertile men. Purpose of the work: analysis of the biochemical composition of sperm plasma, spermogram and sperm morphology in subfertile men of reproductive age. The study included 85 patients aged 25 to 45 years, who were under observation in the clinic of Professor M.A. Florova (Samara): 60 patients seeking help to optimize reproductive health and procreation planning (a group of subfertile men, spermogram indices have deviations) and 25 clinically healthy men - a comparison group (control, spermogram indices are within the WHO norm). In subfertile men, morphological abnormalities of spermatozoa and changes in the biochemical composition of sperm plasma (increased concentration of transamination enzymes) were established. Spermatozoa with immature chromatin were identified. Electron microscopic analysis revealed fragmentation of sperm DNA: the highest percentage of spermatozoa with fragmented DNA was found in subfertile men (more than 22%).

The use of autologous adipose tissue-derived cell products to restore the morphology and function of organs and tissues affected by the disease have become widespread in regenerative medicine. For the wide application of cell-based products in clinical practice, it is important to develop and implement new devices to automate and standardize the isolation of stromal vascular fraction cells. This work aimed to compare cell products obtained from human adipose tissue using two systems, both registered in the Russian Federation and based on enzymatic and mechanical methods of cell fractions isolation. As clinical material, we used samples of adipose tissue in the form of lipoaspirate obtained from healthy patients. The isolation of cell products was performed according to the instructions for the use of these systems. A comparative analysis of the effectiveness of these systems for the isolation of cell fractions was carried out by evaluating several parameters determined for obtained cell products. The cell product obtained by the enzymatic method is characterized by a high yield of nucleated cells, as well as a high proliferative potential of stem cells isolated from adipose tissue. The cellular product obtained from adipose tissue using the mechanical method is characterized by a low yield of nucleated cells, the presence of a large volume of residual oil, and destroyed connective tissue in the final product. The work shows that the method of adipose tissue processing (enzymatic or mechanical) has a significant effect on the characteristics of the products obtained.

There is a strong unmet need for bone grafts and substitutes combining osteoinductive capacities and biophysical properties for using them in minimally invasive surgical interventions. We have developed three variants of injectable gene-activated bone substitutes containing three components: octacalcium phosphate microgranules (OCP), plasmid DNA delivering the gene of vascular endothelial growth factor, and one of the hydrogels based on sodium alginate, type I collagen, and hyaluronic acid. The molecules of the gene constructs were contained both in the hydrogel and on the surface of the OCP microgranules. In the model of a critical-sized bone defect in rabbit parietal bone, we found that all the gene-activated hydrogels contributed to bone tissue regeneration, however, the largest amount of newly formed bone, including those in the central part of the defect, was detected in the groups with gene-activated materials based on hyaluronic acid and collagen. Thus, the developed materials can be considered as candidates for medical devices, but additional studies are required to assess the dose-depended effect and optimize the materials composition.