Проведено доклиническое изучение безопасности лекарственного средства на основе комплекса лития цитрата, полидиметилсилоксана и оксида алюминия. Однократное его введение внутрижелудочно в максимально возможной дозе (12000 мг/кг) мышам и крысам не вызывало гибели животных. В хроническом эксперименте на крысах при введении этого препарата в дозах, значительно превышающих рекомендуемые для человека (4000 мг/кг), внутрижелудочно в течение 90 дней не наблюдалось изменений массы тела относительно крыс контрольных групп, не выявлено токсического действия препарата на функциональную активность и морфологическое состояние основных органов и систем (ЦНС, кровь и костный мозг, печень, почки, сердце, желудочно-кишечный тракт, половые органы).

A comparative study of the effect of a sorbent with nanotubes (Al2O3@ WCNT-PDMS) and a carbon-mineral sorbent (Al2O3@C) on the parameters of human erythrocytes was carried out. Using scanning flow cytometry, the morphological and functional parameters of venous blood erythrocytes as well as drainage blood after its perfusion through columns with sorbents were determined. The compared samples Al2O3@SWCNT-PDMS and Al2O3@C are similar by their effect on the morphological and functional parameters of erythrocytes. The maximum membrane extensibility increased to a greatest extent after contact with Al2O3@C, the amount of hemoglobin in erythrocytes decreased to the greatest extent after perfusion through a column with Al2O3@SWCNT-PDMS sorbent. The scanning flow cytometry is promising for assessing the effect on erythrocytes of new sorption materials intended for blood detoxification. Changes in the parameters of erythrocytes of blood collected in a sterile drainage system for subsequent reinfusion were revealed.

It is known that the circadian rhythm of melatonin production depends on the intensity of illumination. Violation of the light regime leads to suppression of melatonin synthesis and the development of desynchronosis, which increases the risk of developing a number of pathologies. In this regard, it is relevant to search for opportunities to restore disturbed circadian rhythms and, especially, to correct immune dysfunctions that occur in these situations.The aim of this study was to examine the effect of a complex of melatonin, aluminum oxide and polymethylsiloxane on the lymphocytes of the spleen of mice kept under round-the-clock lighting.Materials and methods. Mice of the C57Bl/6J line were kept under round-the-clock lighting for 14 days, against which they were intragastrically injected with distilled water, aluminum oxide with polydimethylsiloxane, melatonin and a complex of melatonin, aluminum oxide and polymethylsiloxane (a new drug developed by the Research Institute of Clinical and Experimental Lymphology – Branch of the Federal Research Center Institute of Cytology and Genetics SB RAS; Patent of Russian Federation No. 2577580, 2016), represented by a complex of porous material (aluminum oxide with polydimethylsiloxane) and melatonin, immobilized in the pores, from which it is gradually released in a liquid medium. Intact animals kept under the light regime of ST 12/12 and under round-the-clock lighting served as a control. Immunophenotyping of spleen B- and T-lymphocytes was performed on a flow cytofluorimeter with monoclonal antibodies APC CD3 and FITC CD19. For studying the distribution of cells by stages of the cell cycle in splenocytes, the amount of intracellular DNA was measured by the level of inclusion of propidium iodide.Results. Flow cytometry of the distribution of B- and T-lymphocytes of the spleen in male mice of the C57Bl/6J line kept under round-the-clock lighting conditions (KO 24/0 h) revealed a decrease in the percentage of B-lymphocytes and an increase in the number of T-lymphocytes, compared with animals kept under standard lighting conditions (the light/dark photoperiod – 14/10 hours). The ratio of CD19+/CD3+ lymphocytes of the spleen in mice under the conditions of KO significantly decreases (1.5 times) compared to intact animals (p ≤ 0.001). The administration of pure and modified melatonin (Complex M) to animals kept under round-the-clock lighting conditions has an equally pronounced normalizing effect on the cellular composition of B- (CD19) and T- (CD3) lymphocytes of the spleen, bringing the values of the studied parameters to the control values of the intact animals (p ≤ 0.001) Round-the-clock lighting affects the proliferative potential of splenocytes, reducing the number of cells in the G2/M phase, compared with animals treated with melatonin (p ≤ 0.050). The introduction of melatonin leads to an increase in the percentage of cells in the G2/M phase relative to the placebo group (p ≤ 0.050). In the group of mice treated with Complex M, the greatest increase in cells at the S + G2/M phases and the highest percentage of cells at the G2/M phase were revealed compared to the placebo control group (p ≤ 0.050).Conclusion. The complex of melatonin, aluminum oxide and polymethylsiloxane has additional immunotropic properties in relation to the modifier molecule, which, apparently, are due to the joint immunostimulating effect of melatonin and the lymphostimulating effect of the sorbent. Melatonin in the composition of the complex shows its properties more stably.

The use of lithium drugs in clinical practice requires constant monitoring of lithium plasma concentration, because toxicity is sometimes observed at therapeutic concentrations of lithium. This is often associated with fluctuations of plasma concentration of lithium ions after intake of individual doses. Therefore, the use of a porous carrier providing a stable blood level of the drug is extremely promising and important for clinical practice. We studied activity of a new lithium drug (lithium complex) consisting of aluminum-silicon base and lithium citrate immobilized on its surface. Lithium carbonate served as the reference drug. It was shown that lithium carbonate and lithium complex exhibited no anxiolytic activity in the conflict model, but produced an antidepressant effect and improved exploratory behavior of animals.

In modern-day conditions, the prevalence of situations in which light disrupts one’s daily life has increased. This causes a decrease in the synthesis of melatonin at nighttime. Melatonin affects the body’s lymphocytes differentiation, inflammation and immune responses. Therefore it is important to study the effect of light disturbance on the organs of the immune system as well as the possibilities of correcting immunity disorders with the help of a new melatonin-containing drug (Complex M), synthesized in the laboratory of pharmaceutical technologies of Research Institute of Clinical and Experimental Lymphology (RICEL), a branch of the Institute of Cytology and Genetics (ICG). The purpose of this study is to identify the morphological features of the thymus and spleen of mice after intragastric administration of the Complex M within 7 days alongside round-the-clock illumination. The experiment used male C57Bl/6J SPF mice aged 10-12 weeks. The day after the last administration of Complex M, the animals were sacrificed by cranio-cervical dislocation and the thymus and spleen samples were taken. The spleen and thymus samples were fixed by immersion in 10% buffered formalin and enclosed in Histomix. Sections were stained with hematoxylin and eosin and morphometric analysis was performed using ImagePro and Image J programms. The administration of Complex M led to an increase in the diameter of the pulp veins and an increase in the cortico-medullary index of the thymus, which indicates its activating effect on the central differentiation of T-lymphocytes.

Экспериментальное изучение токсического влияния

Circadian variations in the cellular composition of the lymphoid organs were studied in female Wistar rats under normal conditions and in experimental endomyometritis. The fractions of CD8(+) cells (effector killers), CD25(+) cells (activated/immature lymphocytes), as well as large, medium, small lymphocytes and monocytes/macrophages were assessed at 10.00 and 20.00 h. In the thymus and spleen of rats with endomyometritis, the number of parameters demonstrating significant circadian variations was lower than in intact animals. In the lymph nodes, morning/evening differences appeared for the number of CD8(+) and CD25(+) cells and monocytes/macrophages in the para-aortic lymph nodes, the number of large and small lymphocytes and CD8(+) cells in inguinal lymph nodes, and in the number of large lymphocytes, CD8(+) cells, and monocytes/macrophages in the ileal lymph nodes. Thus, the development of chronic inflammation in the uterine and vaginal mucosa was accompanied by desynchronosis in the immune system. Hence, circadian rhythms should be taken into consideration in the diagnosis and treatment of the disease.

Research Institute of Clinical and Experimental Lymphology has developed an innovative drug based on a complex of lithium citrate, polymethylsiloxane and aluminum oxide (LOAP). Lithium-based drugs are effective in treating bipolar disorders. However, the toxic effects of lithium cause a “narrow therapeutic window”, which limits its clinical use. The creation of the drug LOAP was aimed at creating a prolonged form with a slow release of lithium to reduce toxic properties and use lithium citrate as an active pharmacological agent. At the moment, the lithium complex has no analogues. The purpose of the study was to study the parameters of acute toxicity of the LOAP. Material and methods. When studying acute toxicity, drugs were administered once intragastrically to mice and rats at doses of 12000, 10000, and 5000 mg/kg. Results. A single administration of drugs intragastrically through a probe in the maximum possible doses to mice and rats did not cause the death of animals and did not cause a locally irritating effect on the gastric mucosa. LOAP can be assigned to hazard class 4 (GOST 12.1.007-76).

The cell composition of leukocyte infiltrates in the endometrium, myometrium, and vaginal walls was studied in Wistar rats with modeled chronic endomyometritis after administration of IFNγ (0.1 μg/100 g body weight) in different daily regimens (10.00 or 20.00). Morning injections of this cytokine ameliorated inflammatory infiltration of the uterine wall and vagina, but increased the content of neutrophils in the endometrium. Evening cytokine injections reduced neutrophilic infiltration, enhanced mononuclear infiltration, and had no effect on plasmacytic infiltration of the uterine and vaginal walls. In the vaginal wall, both IFNγ administration schedules decreased neutrophil content. The data indicate the necessity to take into account the circadian rhythms in IFN therapy.

Research Institute of Clinical and Experimental Lymphology has developed an innovative drug based on a complex of lithium citrate, polymethylsiloxane and aluminum oxide (LOAP). Lithium-based drugs are effective in treating bipolar disorders. However, the toxic effects of lithium cause a “narrow therapeutic window”, which limits its clinical use. The creation of the drug LOAP was aimed at creating a prolonged form with a slow release of lithium to reduce toxic properties and use lithium citrate as an active pharmacological agent. At the moment, the lithium complex has no analogues. The purpose of the study was to study the parameters of acute toxicity of the LOAP. Material and methods. When studying acute toxicity, drugs were administered once intragastrically to mice and rats at doses of 12000, 10000, and 5000 mg/kg. Results. A single administration of drugs intragastrically through a probe in the maximum possible doses to mice and rats did not cause the death of animals and did not cause a locally irritating effect on the gastric mucosa. LOAP can be assigned to hazard class 4 (GOST 12.1.007-76).

High rates of morbidity given a number of pathologies are associated with the accumulation of toxic agents in the body, which is the result of impaired metabolism alongside pre-existing conditions and medication overdoses. The use of porous sorbent materials in medicines allows the removal of toxins from the body, while the active ingredients work to reach the intended outcome. The purpose of this study is to detect the influence of the Al 2 O 3 @PDMS aluminum-silicon sorbent, by scanning flow cytometry, on the morphofunctional parameters of red blood cells during haemoperfusion of blood through the sorbent column. The study of the physical and chemical properties of the porous aluminum-silicon sorbent developed at Research Institute of Clinical and Experimental Lymphology - Branch of the Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences (RICEL - Branch of IC&G SB RAS) was carried out using generally accepted methods for determining the porous structure and adsorption activity. The biological properties of the sorbent were evaluated by its effect on the red blood cells of the donor blood during hemoperfusion of blood through a column with the sorbent. Using scanning flow cytometry, morphological and functional parameters of erythrocytes were determined. Parameter values for populations of 6,000 red blood cells were expressed as an average value ± standard error (Origin 2017 software). The study revealed that the aluminum-silicon sorbent minimally changes morphological parameters, the ratio of stiffness to plasticity of the membrane, and does not critically reduce the number of band 3 proteins in red blood cells responsible for anion exchange of cells, which indicates its safety.

Проведено исследование эмбриотоксического действия нормотимического лекарственного средства на основе комплекса лития цитрата, полиметилсилоксана, оксида алюминия (далее — комплекс лития) в антенатальном и постнатальном периодах развития. Установлено, что в результате применения комплекса лития в дозах 400 и 2000 мг/кг 1 раз в день внутрижелудочно с 1-го по 19-й и с 6-го по 20-й день беременности у крыс выявляется ряд эмбриотоксических эффектов, зависящих от дозы препарата и проявляющихся как в антенатальном, так и постнатальном периоде. В антенатальном периоде развития потомства животных, получавших комплекс лития в течение всего периода беременности, выявлено повышение частоты гибели эмбрионов и плодов до имплантации на (17,5 ± 3,1) % (доза 2000 мг/кг), а после имплантации — на (26,6 ± 5,2) % (доза 2000 мг/кг) и на (15 ± 2,3) % (доза 400 мг/кг), увеличение числа плодов с кровоизлияниями в коже на 12,4 % (доза 2000 мг/кг) и патологическими изменениями внутренних органов в среднем на 50 % (доза 2000 мг/кг) и на 30,7 % (доза 400 мг/кг). В постнатальном периоде развития потомства животных, получавших комплекс лития на 6 – 20-й дни беременности в дозе 2000 мг/кг, выявлено снижение индекса выживаемости крысят на 57,7 %, замедление у них скорости физического развития на 32,4 % и формирования сенсорно-двигательных рефлексов на 21,7 %, снижение способности к обучению и адаптивному поведению на 46,9 % (p < 0,05) Таким образом, препарат обладает дозозависимой эмбриотоксичностью средней степени выраженности, проявляющейся в анте- и постнатальном периоде развития потомства.

Представлены результаты исследования специфической активности лекарственного средства на основе комплекса лития цитрата, полиметилсилоксана и оксида алюминия (далее по тексту комплекс лития) по его адаптогенному действию и влиянию на агрессивное поведение. Установлено, что изучаемый комплекс лития, по сравнению с карбонатом лития, более благоприятно влияет на адаптацию животных к физическим нагрузкам и социальную адаптацию в среднем на 35,9 и 12,1 %, соответственно (p < 0,05), обладает более выраженным антиагрессивным действием на 14,3 % (p < 0,05), не угнетает ориентировочно-исследовательское поведение (угнетение на 105 % (p < 0,05) в случае лития карбоната и на 0 % в случае комплекса лития) и в меньшей мере усугубляет последствия перенесенного острого иммобилизационного стресса на 11,1 % (p < 0,05). Комплекс лития отличается от лития карбоната большей безопасностью.

The purpose of this work was a comparative study of the particle sizes of commercial preparations of colloidal silver by the method of electron microscopy, in connection with the characteristics of specific antimicrobial activity of the preparations under the same conditions. Four commercial preparations of colloidal silver were investigated: Vitargol Forte (manufactured by NPC «Elusan»), Protalor (manufactured by OOO «Esko-Farm», Armenia), Sialor (manufactured by PFK «Obnovlenie»). Protargol (prescription drugstore) served as the drug of comparison. Material and methods. The silver particle sizes in the preparations were determined by high-resolution transmission electron microscopy using a JEM 2010 electron microscope (JEOL, Japan) with a resolution of 0.14 nm at an accelerating voltage of 200 kV. The antimicrobial activity of the silver protein preparations was studied by a standard serial dilution method in liquid nutrient media and diffusion method using bacterial cultures of Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa. Results and discussion. Vitargol Forte, manufactured using advanced technology, is the most fine-dispersed and monodisperse preparation. The preparation of standard classical Protargol, prepared in pharmacy conditions, was the most coarsely dispersed and polydispersed. The conclusion. The dependence of antimicrobial activity of the preparation on the size of its silver particles was shown. Of the four investigated preparations of silver proteinate, the smallest particle size was in the Vitargol Fort preparation. The same drug showed the highest bactericidal and bacteriostatic activity.

Clinical practice and experimental studies have shown that the states of elevated anxiety and depression are often accompanied by impairments to immunity. Investigations over many years have shown that chronic social stress induced by repeated experience of social defeats in daily intermale confrontations leads to the formation of mixed anxiety/depression disorder in mice, which is accompanied by the development of immunosuppression, apparent as a decrease in overall resistance, impairments to humoral and cellular immunity and to proliferation and apoptosis processes in immunocompetent organs, and intensification of oncogenic processes. Primary treatment using anxiolytics and antidepressants to correct psychoemotional status was found to be more effective in the complex correction of psychoneuroimmune impairments than attempts to restore immune measures with the aim of producing therapeutic influences on anxious-depressive states and immunity. A model is proposed for studies of the mechanisms of psychogenic immunodeficiency induced by chronic emotional social stress.

The aim of the study was to study the effect of the normotimic drug based on a complex of lithium citrate, polymethylsiloxane and alumina on DNA damage level (DNA comet test) in vivo by alkaline gel electrophoresis. It was shown that at 3 and 18 hours after the drug administration at the highest dose (2000 mg/kg) and therapeutic dose (400 mg/kg), the percentage of DNA in the tail in the cells of the bone marrow does not differ from the negative controls. That may indicate absence of possible carcinogenic action of the lithium-containing drug. On the other hand, the amount of DNA damage in other organs (kidney, liver, large intestine) increased with an exposure time of 3 hours after administration of the drug in a toxic dose. Although, the degree of this effect expression was not high. When the dose was reduced (400 mg/kg) and the duration of the experiment was increased (up to 18 hours), this effect was not detected. That indicates the ability of the drug to enhance DNA damage repair. Given that lithium preparations have a pronounced toxic effect on kidneys, gastrointestinal tract, liver, the increase in the percentage of DNA in the tail in these organs can be a consequence of the cytostatic action of the drug.

This review summarizes the literature data on the role of lithium compounds in modern pharmacotherapy of various diseases of the central nervous system. Attention is also paid to other therapeutic properties of lithium in atherosclerosis, cardiovascular diseases, diabetes, hematopoietic disorders, inflammation, and diseases of the urinary system. Possible ways of delivering lithium into the body have been charted, in particular, when lithium salt is combined with a sorbent (solid porous carrier). Such compounds have additional therapeutic properties. Data on the significance of lithium compounds in studies on models of diseases of the nervous system in animals are analyzed. Among these models, models of neonatal ischemia/hypoxia of the brain in vivo, neurodegenerative diseases, psychopathological states (aggressiveness, depression) and craniocerebral injury are discussed. There are researches in which the results of the lithium preparations use in clinical practice are investigated. It emphasizes the influence of genetic factors on the lithium effects. Particular attention is paid to the possibility of preventing the toxicity of lithium compounds for the body. The currently known molecular mechanisms of lithium action are discussed: inhibition of glycogen synthase kinase 3β (GSK-3β) and inositol monophosphatase 1 (IMPA1), which have key value for autophagy, oxidative stress, inflammation, mitochondrial function, induction of neurotrophic factors, apoptosis. It was concluded that the study of the molecular pathways of the functioning of lithium compounds empowers understanding both the reasons for its effectiveness in the nervous system diseases and the mechanisms of action on other body systems.

Data on synthesis and research of physical and chemical and biological properties meso - macroporous carriers of different particle size distribution (0.1 mm and 0.2-0.8 mm) on the basis of mechanically strong porous oxide of aluminum ( γ-A1203) and silica polymer (polydimethylsiloxane) are provided in this study. The carriers (white color) received at temperatures up to 200°C have the hydrophilic and hydrophobic nature of a surface and size of a specific surface to 200 m 2 /g. Adsorptive properties of carriers are estimated: on dye of methylene blue 5.5 mg/g (for particles with size 0.1 mm) and 6.9 mg/g (for particles with size 0.2-0.8 mm) and on B12 vitamin 0.6 mg/g for small and large size particles. It is shown, that carriers does not inhibit proliferative potential of human bone marrow mesenchymal stem cells and endothelial cell line EA.hy 926. Carriers are not toxic, it was not succeeded to define the LD50 on the basis of the carrier and lithium citrate (0.5%), not observed the death of animals at the doses, by 30-times exceeding a therapeutic dose and no have observed chronic toxicity (in rabbits, and in rats). Carriers are perspective for creation of the drugs containing active ingredients, such as lithium, melatonin and silver.

We studied the effects of intravenous and lymphotropic administration of bone marrow multipotent mesenchymal stromal cells and products secreted by these cells into conditioned medium on the blood and lymph circulation in the uterus and ovaries, as well as on folliculogenesis in female Wistar rats. It was shown that stromal cells and conditioned media of these cells administered via both routes lead to an increase in the number and diameter of blood vessels in the uterine wall and in the cortical layer of the ovaries. Neither mesenchymal stromal cells, not conditioned media affected the ovarian follicular apparatus.