Vol 15, No 3 (2020)

The historical article presents material that analyzes the contributor from the first female histologists V.M. Danchakova in the study of problems of evolutionary histology and questions of normal and pathological histogenesis. Vera Mikhailovna Danchakova is a student of Professor A.A. Maksimov, the first woman, who defended a histological dissertation at the Imperial Military Medical Academy (1907). In foreign literature, her work is referred to as pioneering in the study of cells, which are now called stem cells. V.M. Danchakova made a significant contribution to the study of the evolutionary aspects of histogenesis of tissues with various cambial properties, for the first time showed the extragonadal origin of primary germ cells using the example of birds, and obtained new additional experimental data confirming A.A. Maksimov's unitary theory of hematopoiesis. Thanks to her research, the possibility of malignancy of stem and progenitor cells was shown for the first time. V.M. Danchakova was prompted by the study of generative cells (primary germ cells, gonocytes), as well as the influence of male and female sex hormones on sex differentiation in embryo-genesis. The results of her experiments using the method of tissue and cell cultures formed the basis for the development of the doctrine of the immune response "graft versus host”.

Atherosclerosis is one of the key causes of morbidity and mortality worldwide. It is known that a leading role in the development and progression of atherosclerosis is played by a violation of lipid metabolism. ABC transporters provide lipid cell homeostasis, performing a number of transport functions - moving lipids inside the cell, in the plasma membrane, and also removing lipids from the cell. In a large group of ABC transporters, about 20 take part in lipid homeostasis, playing, among other things, an important role in the pathogenesis of atherosclerosis. It was shown that cholesterol is not only a substrate for a number of ABC transporters, but also able to modulate their activity. Regulation of activity is carried out due to specific lipid-protein interactions.

Varicocele is a disease that is diagnosed in approximately 10% of males' population. There is data of varicocele which consider the condition as one of the most common causes of male infertility. It is only surgical treatment, to date, that is effective one. However, there is still a group of patients whose fertility problem have not been resolved with the varicocelectomy, that have led scientists to find new ways for treating male infertility. Regenerative medicine and platelet-rich plasma (PRP), in particular, have recently grown up rapidly worldwide. Many preclinical and clinical trials have already demonstrated PRP-therapy effectiveness and safety, despite the novelty of the method. This is why the interest in use PRP-therapy for treating male infertility has increased.

One of the causes of infertility is endometrial hyperplasia. The treatment of it is a prerequisite before the in vitro fertilization. The aim of the study was to develop measures to prevent the development of endometrial hyperplastic processes in patients who have undergone the in vitro fertilization. The paper discusses the results of hysteroscopic, immuno-histochemical, and histological examinations of the endometrium of 179 infertile women with hyperplasia of the uterine mucosa before the ovulation stimulation in the in vitro fertilization protocols and after the protocols. The results were compared with similar indicators of 119 fertile patients with endometrial hyperplastic processes. Significant differences in the expression of endometrial receptivity were obtained. Immunohistochemical analysis indicated a significant decrease in the level of α-2 microglobulin of fertility with preserved estrogen and progesterone receptions even with a normal histological result. Based on the results of the study, an algorithm for the rehabilitation of patients after unsuccessful attempts at in vitro fertilization is proposed. It consists of three stages and allows to restore normal morphological and functional indicators of the endometrium, which is important for the subsequent implementation of the reproductive function. With a personalized approach to hormone therapy, complete restoration of endometrial function in infertile patients is achieved in 64% of cases. It was found that with an adequate reaction of the body to the therapeutic effect, endometrial reception is restored regardless of the presence or absence of fertility. As a prognostic criterion for pregnancy, it is possible to use indicators of α-2 microglobulin of fertility in the menstrual blood and ultrasound parameters of the endometrium in various phases of the menstrual cycle.

Gonadotropins have been used for the last 50 years in assisted reproductive technologies. Often, due to hyperstimulation, there is a worsening effect on both eggs and patients. Therefore, it is possible to use the swine oocyte maturation system in vitro as a culture model for testing hormones used in humans. The purpose of our research was to study the possibility of using human chorionic gonadotropin for maturation of swine oocytes in vitro. The criterion of successful maturation was the presence of the first polar body (MII stage), as well as the ability of matured oocytes to reach the blastocyst stage after artificial (parthenogenetic) activation. It is shown that in the medium with Ovogest(human chorionic gonadotropin derived from urine) there is a tendency to improve the maturation of the nucleus compared to the variant with Pregnyl(human chorionic gonadotropinderived from urineandused for "human” in vitro fertilization) - 59,5% vs 45,8%. After artificial activation, the oocytes reached the stage MII, the cleavage is approximately the same in both cases (89,4% vs 81,2%). But, the percentage of blastocysts obtained, both from the total number of activated eggs, and from the number of cleaving after activation, was greater in variant with Pregnyl(42,4% vs 23,4% without significant difference and 51,2% vs 26,2% with significant difference р<0,05, respectively). Our results show that human chorionic gonadotropin, used for "human” in vitro fertilization, can be used for maturation of swine oocytes in vitro, which is confirmed by the successful use of eggs in experiments on parthenogenetic activation of embryos. It is also possible to use the swine oocyte maturation system in vitro as a culture model for testing hormones used in human in vitro fertilization programs.

Hypertrophic cardiomyopathy is one of the most common cardiovascular pathologies. In most cases, the disease is caused by mutations in genes encoding for sarcomeric proteins. However, high genetic heterogeneity of hypertrophic cardiomyopathy makes it difficult to interpret results of patients' genetic studies. The aim of this study is to check if hypertrophic cardiomyopathy in 15 patients suffering from the disease is due to genetic causes. In the course of genetic analysis, a known pathogenic mutation p.Gln1233Ter in MYBPC3 causing hypertrophic cardiomyopathy was found only in one patient. In six patients, mutations with uncertain clinical significance were identified in hypertrophic cardiomyopathy-associated genes LDB3, MYBPC3, MyH7, MYL2, and MyPn. Three of the mutations, p.Ile730Asn in LDB3, p.Asn515del in MYBPC3, p.Arg955Trp in MYPN were found for the first time in association with hypertrophic cardiomyopathy. In two patients, novel mutations, p.Ser478Trp in MyBpC3 and p.Asn989Ile in MYPN, were identified. Thus, hypertrophic cardiomyopathy may be accounted for by genetic causes in 8 patients more but the role of these mutations in the disease development needs to be clarified.

Transplantation of various types of stem cells as a possible therapy for stroke has been tested for years and the results are promising. Recently, most researchers are inclined to assume that the therapeutic effect of stem cell therapy is based on the mechanism of paracrine action associated with the secretion wide set of regulatory proteins. The aim of this study was to evaluate therapeutic effects of iPSC-derived glial progenitor cells conditioned medium in the rat middle cerebral artery occlusion model of the ischemic stroke. We showed that intra-arterial administration of glial progenitor cells conditioned medium promoted faster decrease of neurological deficit compared to the control group. Moreover, expression of gap43, bax, and tnfa genes involved in neuritogenesis, apoptosis and neuroinflammation was altered. However, no significant enhanced reduction of the infarct volume was registered. Our results demonstrated that administration of glial progenitor cells conditioned medium induced functional recovery after experimental stroke and may affect brain plasticity.

The rare occurrence, high histological heterogeneity, changing classification standards, and complexity of cultivation are the reasons for the failure of soft tissue and bone sarcomas (STBS) cell lines required for large-scale preclinical studies. Obtaining and characterization new STBS cell lines is the great importance for creating cell models that allow us to study the processes of oncogenesis and metastasis of mesenchymal tumors in the development of new treatment methods. Purpose: to create a collection of new characterized STBS cell lines suitable for translational research. In the study were used tumor samples from 71 patients of the N.N. Petrov National Medical Research Center of Oncology with the diagnosis of "sarcoma”. The samples were received during the operation in period 2013-2020. A standard protocol was developed for the cultivation of STBS cells. The proliferative, invasive and migration activity of the obtained cell lines in monolayer and spheroids was studied, HLA typing was performed, and the expression of cancer-testicular genes (CTG) and stem cell markers (CD133 and ALDH1). A collection of STBS cell lines, which includes 54 stable cell lines of 18 histological subtypes, including 39 soft tissue and 15 bone sarcomas cultures, with the metastatic culture of 81.5% was created (n=44). In the process of long-term cultivation there was an increase in the proliferative activity of sarcomas cells (p<0,05). Statistically significant differences in migration activity parameters between soft tissue and bone sarcomas cultures were revealed (p<0,05). A high degree of heterogeneity of the transcriptional activity of the studied CTG was found, and the absence of expression was detected in 19.2% of cases. The activity of the PRAME (55.8%) and GAGE1 (50%) genes was detected with the highest frequency (rho=0.5025; p=0.00015). PASD1 expression correlated with GAGE1 (rho=0.6951; p=0.00001), PRAME (rho=0.5743; p=0.00001). Co-expression of the NY-ESO-1 and MAGEA1 genes was also detected: (rho=0.4027; p=0.00308). The inverse correlation of average strength between the number of ALDH1 + cells and CD133+cells in cultures and the value of time to progression in patients (rho=-0.505, p=0.033; rho=-0.513, p=0.021, respectively) was established. The received and characterized cell lines can become an important component for realization of perspective translational research, a demanded tool for solution of various tasks of modern medicine, such as proteomic research, revealing of important signal mechanisms, search of target molecules for creation of new therapeutic approaches.

Dental implantation in the area of previously lost teeth is a modern way to rehabilitate edentulous patients. The search for osteoplastic materials and methods of preimplantation osteoplasty remains topical. One of these materials is an autogenous dentin matrix (ADM), which is based on a crushed extracted tooth. ADM production and application technologies are different and insufficiently tested. Nevertheless, the application of this technique may open up new clinical possibilities. The study on the use of ADM involved 20 patients from 18 to 65 years old. The patients underwent complex surgical and orthopedic treatment to restore chewing function. At the first stage, the extraction of teeth was performed with the simultaneous preservation of the socket of the extracted tooth ADM. The second stage was carried out 3-4 months later, which included the collection of bone biopsy specimen and subsequent dental implantation in the area of the previously extracted tooth. 42 specimens obtained 12-17 weeks after removal were subjected to histological examination. According to its structure, the bone tissue formed in the hole is reticulofibrous bone. Which allows us to conclude that by the time of 15-17 weeks, the processes of active reparative bone formation in the hole are completed, and the necessary local conditions for dental implantation have been created.

The growing impact of assisted reproductive technologies on the quality of human life and health has required the leading countries to establish a legal framework in the sphere of biomedical research. The question of the legitimacy of genetic modification of human embryos for reproductive purposes is particularly acute. The public and the scientific community give a controversial ethical assessment of the prospects for human DNA editing to new generations. This fact arouses interest in the legal environment of countries that have achieved some success in the sphere of medical innovation. The People's Republic of China was one of the first countries to declare successful genetic research with human embryos. The purpose of this work is to identify the legal boundaries of research to create inherited modifications of the human embryo genome in China. The methodology used in the research: dialectical method as a universal tool of cognition with method of dogmatic (formal-legal) analysis, discourse analysis and case study. By using these methods managed to structure and meaningfully disclose the regulatory framework of the PRC, regulating genetic research of the human embryo for reproductive purposes. The legal documents regulating preclinical and clinical embryo genome studies are summarized. The particular attention is paid to legal precedent of human DNA editing.