Method for preventing biological membrane damages
SUBSTANCE: erythrocyte cell medium is added with an aqueous solution of sodium and potassium salts of humic acids prepared on brown coal of leonardite in a dose of 10.0 mg/kg. That is incubated at a temperature of 37°C for 40 minutes before treatment with acidic haemolytic.
EFFECT: invention enables normalising the cell membrane permeability and reducing the damaged cell count under acidic haemolytic.
The invention relates to the field of medicine and pharmacology, namely to methods of prevention of damage to biological membranes of red blood cells, and can be used to protect biological membranes of erythrocytes when exposed to chemical hemolytic agents.
It is known that the biological damage of cell membranes is one of the universal pathogenetic mechanisms of various diseases (Vladimirov, Y. A. Role of disturbances in the properties of the lipid layer of membranes in the development of pathological processes // Pathologist. physiology and experimental. therapy. - 1989. - No. 4. - Pp. 7-19; Gennis R. Biomembrane. Molecular structure and function: translation from English. - 1st edition. - M.: Mir, 1997. - 624 p.). In this regard, the development of methods for the restoration of the functional properties of cell membranes may be pathogenetically proven strategy in the prevention and treatment of various diseases (Molecular defects of the membrane of red blood cells in the pathology of different Genesis are the typical reaction of the body: the contours of the problem / Novitsky V. V. [and others] // Bulletin of Siberian medicine, 2006. - No. 2. - P. 62-69). It is known the use in pathogenetic therapy of diseases of medicinal substances that have the ability to protect components from damage of biological membranes of different cells. For example, the known method of treatment of purulent-vos�Alitalia diseases of the maxillofacial area, including the opening of purulent-inflammatory focus, drug exposure, characterized in that intraoperatively or immediately after surgery and then daily once the patient intravenous bolus within 4-5 days entering membranoprotector - drug Rext in physiological solution, at the rate of 0.2 µg/kg body weight of the patient (application RU 2006110681; IPC A61K 38/43, AR 43/00, 2007). Drug Rext is an enzyme superoxide dismutase, which is an endogenous scavenger of oxygen free radicals and prevents oxidative modification of proteins and is associated with activation of lipid peroxidation, the destruction of biomembranes of cells (Reference Vidal, http://www.vidal.ru/poisk_preparatov/rexod.htm).
Known hepatoprotective agent for the prevention and correction of toxic liver damage, according to the description as a hepatoprotector used succinate 2-ethyl-6-methyl-3-hydroxypyridine known as Mexidol. Course application of Mexidol has a membrane protective effect on liver cells, toxic affected by an extended period of tuberculostatics (Pat. EN 2189817; IPC A61K 31/4412, AR 43/00, 2002).
The claimed method of increasing the resistance to hemolysis of red blood cells and restore their damaged forms (patent RU 2162698, IPC A61K 35/18; A61K 38/06, 2001). According to the description method.� resistance to hemolysis of red blood cells and restore their damaged forms is the introduction of hydrophilic natural antioxidants in the blood, and is characterized in that the use of antioxidants carnosine (β-alanyl-L-histidine) or its N-acetyl derivative in a certain range of concentrations.
The claimed method of increasing the resistance of erythrocytes to hemolysis induced by ethanol, by introducing into the suspension of erythrocytes in ethanol drug Galavit, dissolved in physiological solution (patent RU 2315605; IPC A61K 35/18, A61P 25/32, 2008). Galavit (aminodimethylaniline sodium) is an immunomodulatory drug with anti-inflammatory action, the mechanism of action is associated with the ability to influence on the functional-metabolic activity of macrophages (Reference Vidal, http://www.vidal.ru/poisk_preparatov/galavit~22901.htm).
It is known that the combined use of low-intensity laser radiation and complex compounds 5-hydroxy-6-methyluracil with succinic acid in experimental peritonitis increases the acid resistance of erythrocytes to the hemolytic action of hydrochloric acid, when applying this combination is most powerful membrane protective effects (antioxidant Effect and laser therapy on the state of membranes of erythrocytes in experimental peritonitis / Rubilen D. V. [and others] // Medical Bulletin of Bashkortostan, 2009. - Vol. 4, No. 2. - P. 102-106).
Closest to the present invention (the prototype) can� to consider using as a means of having gepatoprotektoram effect of ascorbate lithium with gepatoprotectornoe activity against hemolysis of erythrocytes under the influence of alcohol and having lower toxicity compared to other drugs lithium (patent RU 2351326; IPC A61K 31/375, A61P 43/00, A61P 39/00, 2009). However, according to the description of this patent, and according to the known literature data ascorbate lithium is a psychotropic agent may be used mainly in addiction medicine and psychiatry during chemotherapy.
One group of promising compounds for the development of medical ways to protect cell membranes, are natural compounds - humic substances and salts of humic acids (humates) with multidirectional biological activity (Dry S. A. prospects of application of humic and contains acids in medicine and pharmacy / S. A., Dry, P. V. Kuznetsov // Medicine in Kuzbass. - 2009. - No. 1. - S. 10-14). Features of the chemical structure of humic substances determine their surface-active properties (On the mechanism of antidotal action of humates in relation to the petroleum / V. A. Daurov [et al.] / / bull. CENTER, SIBERIAN DIVISION OF THE RUSSIAN MEDICAL ACADEMY. - 2005. - No. 6 - Pp. 143-146; Stom D. I. Combined effect of petroleum products and of humic substances on Daphnia / D. I. Stom, A. V. Daurov // Siberian ecological journal. - 2004. - No. 1. - S. 35-40), which leads to FPIC�to the competitiveness of solubilisate and are dispersed in a hydrophobic organic substance, increasing their solubility and increasing the hydrophilicity of the surfaces of the cells (Possible mechanisms of biological action of humic substances / Stom D. I. [and others] // Siberian medical journal. - 2008. - No. 6. - P. 76-79). This feature of humic substances suggests the possibility of their influence on properties of lipoproteins and other components of biological membranes.
It is known the use of humic substances and natural substrates containing them, in various branches of medicine.
In dermatology it is known the use of oxidative peat as a means for the treatment of patients with chronic dermatoses (patent RU 2187315; IPC A61K 35/10, A61P 37/04, 20020), the use of products containing sodium HUMATE wood-grass peat deposits "Dark" Tomsk region as a means of having antimicrobial and antifungal activity (patent RU 2340328; IPC A61K 9/06; A61K 35/10; A61P 31/10, 2008). It is known to use the extract of sapropel obtained as interaction between sapropel and pyrophosphate potassium at a weight ratio of 25:1, exhibiting analgesic, anti-inflammatory and resolving the action as a biologically active additive for cosmetic and therapeutic products (patent RU 2170094; IPC A61K 35/00, A61K 7/00, A61K 7/48, 2001). It is known the use of humic acids and humates as a biologically act�VNOM additives in the cosmetic industry, their antioxidant activity (patent RU 2094042; IPC A61K 7/00, A61K 7/40, 1997). You know the way to care for hair and scalp on the basis of humic concentrate, which is a product of processing of peat with a solution of hydroxide of sodium or potassium in the activation of oxygen in the sparging process and subsequent neutralization with a solution of citric or acetic acid, containing on a dry substance of humic acids not less than 2.8 wt.% (patent RU 2394556; IPC A61K 8/97, A61Q 5/12, 2010).
It is known the use of humates in rheumatology, such as electrophoresis 0,09-0,15% solution of sodium HUMATE to improve the effectiveness of rehabilitation treatment of patients with osteoarthritis (patent RU 2196626; IPC A61N 7/00, 2003), a method for the treatment of patients with osteoarthritis, complicated reactive synovitis, including course sapropel mud with a temperature of 35-37°C, magnetic therapy and physical therapy (RU 96117924; IPC A61N 5/06, 1998).
It is known to use mud applications in neurological practice, such as method of rehabilitation treatment of patients with moderate and low risk of recurrent stroke in the period of residual effects of acute disorders of cerebral circulation, including balneotherapy, massage segmental zones, mud baths on the paretic limb (with the use of sapropel lake Kirek to the affected�vechnosti a temperature of 37-38°C for 20 min, a day later, the number of procedures 10) and exercise therapy (patent RU 2009141660; IPC A61H 1/00, 2011). A method of treating patients with persistent neurological manifestations of lumbar osteochondrosis achieved by applying mud applications in the lumbar-sacral region and the affected limb at 40-42°C for 30 min, after which additionally carry out hyponatermia (SU 1711883; IPC A61G 10/02, 1992).
It is known the use of different substrates containing humic substances in gastroenterology. For example, it is known antiulcer effect of mud applications, electrophoresis and peat mud (the Effect of mud applications on the state of acetate-induced ulcer in albino rats / N. N. Auchinawa [et al.] / / Vopr. balneology, physiotherapy and medical physical culture. - 2001. - No. 3. - P. 27-28; experience in the treatment of patients with peptic ulcer of the duodenum SMT-electrophoresis peat mud in a day hospital / Korepanov, A. M. [and others] // Vopr. balneology, physiotherapy and therapeutic physical training. - 2003. - No. 5. - Page 22-25); the use of ulcers in the stomach electrophoresis drug humisolum, which is a biogenic stimulator and representing a 0.01% solution of humic acid fractions Haapsalu sea therapeutic mud of the Baltic sea in isotonic sodium chloride solution (filmed � production in 1997) (G. V. Lurie The effectiveness of different methods of electrophoresis gumizol from the environment Dimexidum with gastric ulcer / G. V. Lurie // Modern aspects of resort gastroenterology: SB. nauch. Tr. - Pyatigorsk, 1988. - P. 46-49). It is known to use by oral administration of the extract of silt therapeutic muds (drug, Palidin, contains a complex of natural biologically active substances) for the treatment of gastric ulcer and duodenal ulcer, the drug belongs to the group of biogenic stimulators (Mashkovsky M. D. Medicines: in 2 vols 1. - 14-e Izd., Rev., Rev. and extra - M.: OOO "Publishing house New wave": the Publisher S. B. Divov, 2002. - P. 174). A method of treating gastric ulcer and duodenal ulcer, characterized in that the medicinal product is obtained by processing of peat, containing a complex of biologically active substances, including humic acid. According to the description of the oral administration of the above medication peat once daily intragastrically at a dose of 50-100 mg/kg provides a complex effect on different pathogenetic links of the development of gastric ulcer, showing both local and overall health benefits (patent RU 2357741; IPC A61K 35/10; AR 1/04, 2009).
A method of treating cancer in the experiment, including�ment chemotherapy of tumors cyclophosphamide and the introduction of funds from peat, containing humic acids, fulvic acids, etc., and a means of peat is introduced into the feed or drinking water based on the daily doses of 100-200 mg/kg (patent RU 2396967; IPC A61K 35/10; OR 35/00, 2010). It is known the use of suppositories for treatment of chronic prostatitis, consisting of 2.0 ml of a 1% aqueous solution of humic acids (pH 7.4) low-mineralized peloids and sufficient cocoa butter to get suppositories weighing 2.0-2.2 g (patent RU 2241471; IPC A61K 35/10, A61K 35/02, A61K 9/02, AR 13/08, 2004).
It is also known the use of humates from peat and sapropel in veterinary medicine, such as prevention of liver diseases in cows, including the introduction in the composition of the diet of humic funds from the peat used in the form of a solution with 1.0% content of humic acids daily once daily at a dose of 0.125-0.25 ml/kg of body weight two courses duration 25-28 days (patent RU 2372926; IPC A61K 35/10, 2009).
However, the known methods of protection of biological membranes of mammals and human beings with the aid of salts of humic acids, in particular salts of humic acids derived from leonardite, not previously reported. The object of the present invention is to expand the Arsenal of known methods of protection of biological membranes of red blood cells in the damaging effects of chemical hemolytic agents.
Technical results�the ATA is in use to prevent damage to biological membranes of erythrocytes of the drug on the basis of low-toxicity, cheap, available natural raw materials.
The technical result is achieved in that a method of preventing damage of chemical hemolytic agents of biological membranes of red blood cells is to use an aqueous solution of sodium and potassium salts of humic acids extracted from brown coal leonardite, administered at a dose of 10.0 mg/kg on Wednesday with cells of erythrocytes and subsequent incubation at 37°C for 40 min before the damaging effects of acid hemolyticum. As an acidic hemolytica used, for example, 0.1 to Η the solution of hydrochloric acid. The method provides a reduction in the rate of destruction of cells under the action of acid hemolytica and reducing the number of destroyed cells.
Conventional title of an aqueous solution of sodium and potassium salts of humic acids, HUMATE of leonardite. Substance powder is produced LLC "HUMATE", Irkutsk, name and designation of the products: "HUMATE" - fertilizer based on humic acids, for agricultural use,
THE 2189-004-41764643-98, pharmacological properties have not been studied earlier, membrane-protective properties are not known.
The advantages of the proposed method of prevention of damage to biological membranes using an aqueous solution of sodium and potassium salts of humic acids extracted from brown coal� of leonardite, are the relative cheapness of the substance, widely available raw materials in large volumes for environmentally safe production of the substance, the possibility of industrial production by (Perminova I. V. Humic substances - a challenge to the chemists of the XXI century / I. V. Perminova // Chemistry and life 2008. - No. 1. - Pp. 50-55, 10), low toxicity (Low toxicity and anti-inflammatory activity of humates isolated from peat and sapropel Tomsk region / R. R. Ismatova, A. U. Ziganshin, S. E. Dmitruk, I. V. Fedko // Kazan medical journal. - 2006. Volume 87, No. 6. - S. 454-455) and the lack of serious side effects, high efficiency.
Fig. 1 presents the results of the influence of HUMATE solution of leonardite (10 mg/kg, 40 min incubation at 37°C) on the indicators of acid hemolysis of erythrocytes.
To prevent damage to biological membranes of erythrocytes when exposed to chemical hemolytic agents used a tool from natural raw materials, which was an aqueous solution of sodium and potassium salts of humic acids from brown coal of leonardite (hereinafter - HUMATE of leonardite).
To study the ability of HUMATE of leonardite to modify the structural and functional properties of biological membranes used modified method of registration of chemical (acid) erythrogram on Terskova I.�., Gitelson I. I. (1957) (Gitelson I. I. Erythrogramma as a method of clinical examination of the blood. / / I. Gitelson, I. A. Terskov. - Krasnoyarsk: publishing house of the Siberian branch of the Academy of Sciences of the USSR, 1959. - 247 p.; Terskov I. A. Method of chemical (acid) erythrogram // I. A. Terskov, I. I. Gitelson // Biophysics. - 1957. - Vol. 11, ISS. 2. - S. 259-266.; Workshop on Biophysics / V. G. Artyukhov [and others] / edited by V. G. Artyukhov. - Voronezh: Publishing house of Voronezh state University, 2001. - Pp. 147-161). It is proved that the erythrocytes are involved in the pathological process not only in hematological diseases, but also undergo serious changes of structure and function in diseases of different Genesis. The choice of the membrane of red blood cells as an object for testing is dictated by the fact that it is inherent in the General principles of molecular organization of plasma membranes. Therefore, the patterns of changes in the structure and function of erythrocyte membranes with a certain degree of correction, can be extrapolated to other membrane systems. In addition, the simplicity of the organization of the erythrocyte gives you the opportunity to study the functional properties of the plasma membrane without interference imposed
intracellular membrane and organelles (Molecular defects of the membrane of red blood cells in the pathology of different Genesis are the typical reaction of the body: the contours of the problem / Novitsky V. V. [and others] // Bulletin Sib�designer medicine 2006. - No. 2. - Page 62-69; Postnov Yu. V Primary hypertension as pathology of cell membranes / J. V. Postnov, S. N. Eagles. - M.: Medicine, 1987. - 192 p.).
The assessment of the ability of HUMATE of leonardite to ensure the protection of biomembranes in the damaging effects were performed by changing the parameters of the chemical resistance of erythrocyte cells to the inducer of acid hemolysis, the experiments were performed on 3-component model system, including: 1 - "modifier" - a solution of salts of humic acids derived from leonardite (HUMATE of leonardite); 2 - "membrane" is a suspension of erythrocyte membranes in rats; 3 - "destructor" - the component that is causing the acid hemolysis of erythrocytes, i.e. of 0.1 H solution of hydrochloric acid.
Influence of HUMATE of leonardite on the change of resistance of cell membranes was evaluated using the method of automatic registration of acid erythrogram based on photometric registration process of hemolysis in time, a technique developed at the Department of Biophysics and biotechnology of the Voronezh state University. The principle of the methodology used is based on the method of registration of acid erythrogram, proposed by I. I. Gitelson and N. And. Terskovym (Gitelson I. I. Erythrogramma as a method of clinical examination of the blood. / / I. Gitelson, I. A. Terskov. - Krasnoyarsk: publishing house of the Siberian branch of the Academy of NAU� the USSR, 1959. - 247 p.; Terskov I. A. Method of chemical (acid) erythrogram // I. A. Terskov, I. I. Gitelson // Biophysics. - 1957. - T. 11, Vol.2. - P. 259-266; Workshop on Biophysics / V. G. Artyukhov [and others]. / Under the editorship of V. G. Artyukhov. - Voronezh: Publishing house of Voronezh state University, 2001. Pp. 147-161).
The principle of the method of registration of the acid resistance of erythrocytes is to photometric registration of the dynamics of the collapse of the cells when added to suspensions of red blood cells 100 ál of 0.1 N hydrochloric acid solution (method of acid hemolysis). Measure the resistance for each of the erythrocyte is the time during which there is its destruction. The data obtained permit to build erythrogramma - kinetic curve, reflecting the time-sequential entry of erythrocytes of different resistance in the phase of hemolysis.
Analytically this process can be represented as an equation of the form:
where G(%) - the number of destroyed erythrocytic cells in percent, t is time, expressed in seconds.
Red blood cells for research was obtained from the blood of white outbred rats male total 30 goals weight 210,0±9,5 g. blood sampling was performed by resection of the distal part of the tail, in each series of manipulation experiments were carried out at the same time of day between 8.30 am and 9.30 hours. Blood gained by gravity into the tube with stabilizer (heparin). For the purpose of obtaining su�getting suspension of erythrocytes blood, stabilized with heparin, centrifuged in a centrifuge laboratory medical (model ARF-8, produced by JSC "TNK "Dastan", Kyrgyz Republic) for 15 minutes at a speed of 3000 rpm with intermediate laundering from the stabilizer and plasma in 0.9% sodium chloride solution. For this purpose, to the residue, containing the cells, was added in 0.9% NaCl solution, gently mixed and centrifuged again under the same conditions, the procedure was repeated 3 times.
Hemolysis of erythrocytes was carried out in cuvettes with outer dimensions of 20×40×10 mm and a working volume of 5.0 ml. measuring the values of light scattering was performed at the filter (No. 5) with the maximum transmittance in the region of 490 nm (at a wavelength of λ=490 nm), since at this wavelength not tested the output of hemoglobin in the incubation medium, and the scattering of the samples, the intensity of which is directly proportional to the volume, i.e. the surface of the erythrocyte and the content in the environment of erythrocyte cells. Thus:
where G(%) - the number of destroyed erythrocytic cells in percent, Vi- the volume of a single red blood cell, N is the number of red blood cells in the environment. Consequently, the yield of hemoglobin - a secondary phenomenon and at the used wavelength (490 nm) has virtually no effect on the recorded signal.
Photometric registration process dynamics induced �of emeliza of erythrocytes was carried out using a spectrophotometer (model PE-5400 V, production "Group of companies Ecros", Russia).
The acid induction of hemolysis was performed by adding in the working cell to 5 ml of a suspension of erythrocytes (in isoosmotic with 0.9% NaCl solution) 100 ál of 0.1 H solution of hydrochloric acid. The choice of this material as an acidic hemolytica due to the stability of the solution during storage and the presence of both ions (H+and Cl-) in the blood plasma. Indicators were recorded every 15 seconds for a 735 seconds.
The dynamics of the process of hemolysis of erythrocytes was recorded immediately after the addition of hemolytica (0.1 M HCl) to the erythrocyte suspension (0.9% NaCl).
Digital data displayed on a graph in the result register S-shaped integral of the kinetic curve (erythrogramma), the form of which reflects the total change in the magnitude of light scattering (τ) in the studied solution in time, i.e. τ=f(t). In the process of hemolysis is the breakdown of red blood cells reaches the maximum value (Vmax) about the middle of the curve.
Analysis of erythrogram was carried out by graphical and mathematical method, the indicators of the structural state of erythrocyte membranes (Gennis R. Biomembrane. Molecular structure and function: translation from English. - 1st edition. - M.: Mir, 1997. - 624 p.) when hemolysis was assessed according to design parameters in accordance with the methodology (Practice� in Biophysics / VG Artyukhov [and others]. / Under the editorship of V. G. Artyukhov. - Voronezh: Publishing house of Voronezh state University, 2001. Pp. 147-161).
Calculated the constant maximum speed of hemolysis (KmaxRel. units) - a parameter that characterizes the proportion of red blood cells, simultaneously entered the stage of hemolysis was determined by the formula (1):
where Kmax(Rel. units) - constant maximum speed of hemolysis; tg α is the tangent of the angle α; α - angle of inclination of the linear part of the curve to x-axis, was determined graphically.
We calculated the relative amount of hemolyzed cells (G, %) at different time points of observation by the formula (2):
where Gn- to�number of red blood cells, involved in the process of hemolysis on n second process, expressed as a percentage, was determined every 5 sec for 735 h; K is the rate constant of hemolysis on the t second process was calculated by the formula (3); TODA.AVG.- the average value of the light scattering in the test sample on the n second process; TSREN.- the average value in the test sample source, i.e., 0 sec.
Determined the relative number of spherocytes (Gsf, %) - indicator of the number of red blood cells (mostly niskalaukaus the population) involved in the initial stage of the process of hemolysis - stage spherocytosis. Determine the maximum number of hemolyzed red blood cells in percent (Gmax, %).
Determined time of the latent period of hemolysis (tlat, sec) - the time period after adding hemolytic agent and prior to the registration phase of hemolysis.
To assess the ability of HUMATE of leonardite to ensure the protection of biomembranes in the damaging effects of acid Gamal�teak used an aqueous solution of HUMATE of leonardite in a dose of 10.0 mg/kg, which was added directly into the experimental cuvette, red blood cells, then the mixture of the suspension of erythrocytes and HUMATE of leonardite were subjected to pre-incubation in a thermostat at 37°C for 40 min, and then actually induced acid hemolysis by adding Wednesday with erythrocytes and HUMATE of leonardite 100 ál of 0.1 H solution of hydrochloric acid. The incubation temperature of 37°C is selected as corresponding to the human body temperature, incubation time of 40 min is chosen as the average time required for admission introduction of medicinal substances in the blood plasma. Dose of HUMATE leonardite 10.0 mg/kg is chosen experimentally based on the results of a series of previously conducted experiments with doses of 1.0, and 5.0, 10.0 and 20.0 mg/kg.
Experiments for the dose of HUMATE leonardite 10.0 mg/kg conducted in 3 series of experiments, the results obtained were subjected to statistical analysis with calculation of mean values. In each series of experiments studies were performed in parallel samples, one of which was the control (erythrocytes + hemolytic) and the second experimental (erythrocytes + HUMATE solution of leonardite + hemolytic).
It is established that the process of hemolysis in the control sample corresponds to the parameters of the standard curve and hemolysis has an S-shape. The process ends by an average of 7 minutes with about�slip the percentage of hemolyzed cells to 84.99±4,810%. With an average latent period of hemolysis (tlat) amounted to $ 5,00±1,000 h, the relative number of spherocytes (GSF,) 0,32±0.800 to% and a constant maximum speed of hemolysis (Kmax) amounted to $ 7,115±0,830 Rel. units (Fig.1).
It is established that when introduced into the medium with cells erythrocytes HUMATE of leonardite in a dose of 10.0 mg/kg with pre-incubation for 40 minutes at a temperature 37,0°C was not observed changes in the index tlatidentified a slight increase in the number of spherocytes GSF0.37%. Significant significant decrease of Kmaxby 63.4%, indicating a decrease in the rate of hemolysis and the decrease in the intensity of the process of destruction of cells. During the period of observation revealed the ability of HUMATE of leonardite significantly reduce the percentage of destroyed cells 270 h process showed a reduction in the number of hemolyzed erythrocytes on 29,26%, 375 h, respectively, 32,63% and by the end of the process of hemolysis on 735 s - reduction total number of damaged cells at 20,45%, which was significantly less compared with the control (Fig.1).
Therefore, on the background of the presence in the environment of HUMATE of leonardite was found to be increasing with time and degree of resistance of cells to the damaging effect of acid hemolytica. The revealed changes in the modus�Ana with the ability of HUMATE of leonardite to cause conformational structure and function of cell membranes, due to the interaction with lipoproteinemia components, resulting in reduced permeability to hydrogen ions and provides increased osmotic resistance of the cell membrane.
Thus, for the first time revealed the ability of HUMATE solution of leonardite in a dose of 10.0 mg/kg with pre-incubation with a suspension of erythrocytes at 37°C for 40 min to normalize the permeability of cell membranes, which leads to reduced speed of destruction of cells under the action of acid hemolytica and reducing the amount of destroyed cells. The established changes demonstrate the ability of HUMATE of leonardite in a dose of 10.0 mg/kg to protect biological membranes of red blood cells, which can be used for the prevention of violations of the integrity of biological membranes of red blood cells in the hemolytic action of chemical agents.
A method of preventing damage of chemical hemolytic agents of biological membranes of erythrocytes, consisting in the use of an aqueous solution of sodium and potassium salts of humic acid derived from brown coal leonardite, administered at a dose of 10.0 mg/kg on Wednesday with cells of erythrocytes and subsequent incubation at 37°C for 40 min before the damaging effects of acid hemolyticum.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to a new intensifier of the antitumour effect, which is a uracil derivative of the general formula (I) or its pharmaceutically acceptable salt. In the general formula (I) X represents a C1-5-alkylene group, and wherein one of methylene groups making an alkylene group is optionally substituted by an oxygen atom; R1 represents a hydrogen atom or a C1-6-alkyl group; R2 represents a hydrogen atom or a halogen atom; and R3 represents a C1-6-alkyl group, C2-6-alkenyl group, C3-6-cycloalkyl group, (C3-6-cycloalkyl)-C1-6-alkyl group, halogen-C1-6-alkyl group or a 5-6-merous saturated heterocyclic group with an oxygen atom as a heteroatom, a uracil derivative presented by the following formula (I). The invention also refers to a method for potentiating the antitumour action or a method of treating tumours, involving administering an effective amount of a combination of the above uracil derivative or its pharmaceutically acceptable salt and an antimetabolite in an effective amount. The antimetabolite represents an agent specified in 5-fluoruracil (5-FU), potassium tegafur/gimeracil/oteracil (TS-1), tegafur/uracil (UFT), capecitabin, 5-fluor-2'-deoxyuridine (FdUrd) and Pemetrexed.
EFFECT: preparing the intensifier of the antitumour effect.
18 cl, 10 dwg, 10 tbl, 67 ex
SUBSTANCE: invention relates to medicine, namely to methods of purification and health improvement of an organism. For this purpose therapeutic starvation for not fewer than 5 days in case of a 7-day programme and for not fewer than 7 days in case of a 9-day programme is carried out. The duration of a recovery period constitutes two days. Food intake is realised nine times per day each day both in the process of therapeutic starvation and in the recovery period. In the period of therapeutic starvation the first food intake includes bee products "APIGRANULES 2" in a dose of one teaspoon, "KHINAZI balm" in a dose of one teaspoon, "A-P-V" in a dose of one teaspoon, "APITOK" in a dose of one teaspoon, "Antihelm phyto" - one capsule and still mineral water - one glass. The second food intake includes a drink with ginger, lemon juice, garlic and mint, honey - one teaspoon and one glass of apple-carrot juice. The third food intake supposes an intake of bee products "APIGRANULES 2" in a dose of one teaspoon, "KHINAZI balm" in a dose of one teaspoon, "A-P-V" in a dose of one teaspoon, "Antihelm phyto" - one capsule and still mineral water - one glass. The fourth food intake includes a drink with ginger, lemon juice, garlic and mint, honey - one teaspoon and one glass of apple-carrot juice. The fifth food intake consists of tea black or green with ginger, one teaspoon of honey and one glass of apple-carrot juice. The sixth food intake consists of bee products "APIGRANULES 2" in a dose of one teaspoon, "A-P-V" in a dose of one teaspoon, "Antihelm phyto" - one capsule and one glass of still mineral water. The seventh food intake includes tea black or green with ginger, one teaspoon of honey and one glass of apple-carrot juice. The eighth food intake supposes an intake of tea black or green with ginger, one teaspoon of honey and one glass of apple-carrot juice. The ninth intake of food includes depressant tea, including mint grass, valerian root, fennel seeds, cumin seeds, epilobium or strawberry leaves and one glass of apple-carrot juice. In the recovery period in 1-st, 2-nd, 3-rd, 4-th and 6-th intakes of food the composition of products remains the same as in the process of starvation. For the fifth intake of food the patients take tea black or green with ginger, one teaspoon of honey and vegetable soup. For the seventh intake of food the patients take tea black or green with ginger, "MILK COCKTAIL WITH CHITOSAN". The eighth intake of food includes tea black or green with ginger, one teaspoon of honey, "APICAMPA" cereal. The ninth intake of food for the recovery period includes a baked apple, depressant tea. In addition, a number of procedures are carried out after each food intake during the period of therapeutic starvation and the recovery period. After the first food intake gymnastics "5 Tibetan pearls" is realised. After the second food intake exercises on training apparatuses, procedures of press-therapy, electrolypolysis and myostimulation are realised. After the third food intake exercises of therapeutic physical training or aerobics are performed. After the fourth food intake a rest in form of a walk, a halotherapy session, combined with a session of relaxation therapy are realised. After the fifth food intake a course of strip-plastic is carried out. After the sixth food intake massage by manual application with a peloid-based mixture or manual massage with honey is carried out. After the seventh food intake an infra-red sauna, shower, phytobath with medicinal herbs, honey, ginger, lemon juice is taken. After the eighth food intake a shower is taken.
EFFECT: method ensures effective health improvement of the organism with the preservation of the full value life style and a sense of a comfortable state in the starvation period, reduction of recovery period term after starvation, purification of the organism without loading on the gastrointestinal tract, weight loss, and improvement of the general state and workability of the patients.
3 tbl, 7 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology, specifically to a fused protein containing a variant of rodostomin, and can be used in medicine. An ανβ3 integrin selective polypeptide consisting of an amino acid sequence SEQ ID NO:1 conjugated on the N terminal by a linker amino acid sequence containing a combination of the amino acids glycine and serine with a variant of a human serum albumin (HSA) with SEQ ID NO:4.
EFFECT: invention enables the higher therapeutic effectiveness in the diseases related to ανβ3 integrin.
12 cl, 14 dwg, 2 tbl, 7 ex
SUBSTANCE: invention refers to medicine and can be used for the correction of the individual's functional state and performance. That is ensured by administering Semax neuroactive peptide in a dose of two drops in each nasal passage. That is followed by the electric current exposure covering a frontal-mastoid region at pulse length 0.2ms, current intensity 0.8mA and pulse train 800Hz for 40 min. The exposure is combined with at least 10 sessions of hyperbaric oxygenation at pressure 1.6 atm.
EFFECT: method provides the rapid and effective increase of performance in sportsmen, military men and individuals involved in the other professions related to significant physical and mental stress by improving the functions of various regions of brain cortex as a result of the selected complex exposure enabling the substantial vasodilatation and maximum tissue oxygenation.
SUBSTANCE: using a compound representing a bicyclic pyrimidine derivative as Hrf2 transcription factor activators.
EFFECT: compounds of general formula can be used preventively to increase body defences before operations with toxic chemicals and high radiation doses.
3 cl, 7 tbl, 11 ex, 9 dwg
SUBSTANCE: invention refers to methods for cell and tissue protection against a hypoxic injury and can be used for developing remedies for hypoxic and ischemic injuries. The developed method for protection is based on cell treatment by substances increasing a level of glutathionylation of Na,K-adenosine triphosphatase catalytic subunit that reduces to its activation. The obtained results can be actual for biological research institutions, medical facilities, particularly cardiologic clinics, as well as biotechnological companies for transplantology and tissue engineering.
EFFECT: presented method can be used for research activities aiming at creating the therapeutic agents for relieving tissue injuries, particularly myocardial tissue in hypoxia and hypoxia/reoxygenation.
SUBSTANCE: invention relates to sports medicine and pharmacology, and can be used to increase the overall physical performance of athletes of speed and power sports. To do this, the dietary supplement "Epsorin" is added daily twice a day. At that during the stage of physical training "Epsorin" is taken in for two weeks 30-40 minutes prior to start of the exercise-training process as 10-15 drops per a glass of water; during the period of special physical training it is taken for 10 days 30-40 minutes prior to start of the exercise-training process as 10-15 drops per a glass of water; during the transition period it is taken for 3-5 days 30-40 minutes prior to the training as 5-7 drops per a glass of water; on the day of competitions the dose is increased to 20-30 drops per a glass of water.
EFFECT: method enables to improve the overall physical performance and endurance of athletes due to activation of metabolism and antioxidant systems of the body.
SUBSTANCE: as an ingredient of drinking water or beverage, water containing 1H16OD isotopologue in an amount from 0.0002 to 0.0278 molecular %. The invention provides higher individual's body resistance to the flight factors, such as: height gain, cabin and compartment noise increase, vibration, rolling, humidity disposal, time zone change.
EFFECT: higher resistance of the mammalian body to the effect of the flight factors on an aircraft.
2 tbl, 2 ex
SUBSTANCE: invention refers to medicine, namely to experimental medicine, and can be used for general controlled hyperthermia. That is ensured by the general warm-up of an experimental animal's body with the preliminary intraperitoneal administration of an animal toxin. The intraperitoneal administration of the animal toxin is preceded by administering protamine sulphate in a dose of 10mg/kg; the animal toxin is bee poison 2.5mg/kg, or cobra poison 0.5mg/kg, or carpet viper poison 1.5mg/kg, or copperhead snake poison 2mg/kg, or scorpion poison 1mg/kg, or lebetina viper poison 2mg/kg.
EFFECT: invention provides the higher survival rate of the animals, particularly 24 hours later, with the low negative effect of the administered animal toxin on the animal's body.
2 tbl, 1 ex
SUBSTANCE: conventional integrated therapy involving anti-tussive medications and drugs restoring the bronchopulmonary patency accompanies administering an oral complex immunoglobulin preparation 1 dose of protein 300mg once or twice a day for 5-7 days.
EFFECT: method enables fastening recovery with reducing a rate of complications and improving the clinical outcome.
4 tbl, 1 ex
SUBSTANCE: method involves doing therapeutic exercises and pulsed ultrasonic therapy at a pulse length of 10 ms. That is combined with the magnetic stimulation of the shoulder girdle muscles: m. trapezius m. deltoideus, m. supraspinatus, m. infraspinatus on the involved side. That involves the exposure in accordance with a stable technique with using paired pulses of the length of 220±20 mcs at a pulse repetition frequency of 25±5 pulse/min and a pulse-to-pulse spacing of 80 ms for 3 min per one field. The exposure intensity is 70-90% of the maximum magnetic induction amplitude 1,300 mT. The total length of the procedure is 12 minutes. Then 30 minutes later, mud therapy is conducted by applications on the involved shoulder joint. A mud cake temperature makes 38-40 degrees, and the length of the exposure makes 20 minutes. The mud therapy is followed by ultrasonic therapy. The exposure covers the rotator cuff and attachment points of the tendons - m. supraspinatus, m. infraspinatus, m. teres minor on the involved side. The exposure follows a labile technique at an intensity of 0.2-0.4 W/cm2 for 2 minutes per each field at the total length of the procedure of 8 minutes. The procedures are daily. The therapeutic course is 8-10 procedures.
EFFECT: method enables clinical effectiveness by recovering the motor activity of the involved extremity and prolongs the remission, prevents the progression of the biomechanical and neurodystrophic disorders, and formation of neurological complications.
1 ex, 4 tbl, 1 dwg
SUBSTANCE: invention refers to medicine, particularly to experimental balneology, and concerns a method for preparing concentrated sapropel. A method for preparing concentrated sapropel, involving recovering and concentrating easily disaggregating water-soluble components of sapropel by placing native sapropelinto a vacuum evaporator, which is exhausted and heated; a heating procedure is combined with exposing to ultrasound with measuring an ultrasound rate continuously or intermittently and accelerating ultrasound by differentiating the ultrasound rate on time; the above measurement procedure is repeated until the ultrasound acceleration gets equal to zero; the procedure of recovering the easily disaggregating water-soluble components of sapropel and the concentration procedure is terminated, and a dry residue is solubilised in the concentrated easily disaggregating water-soluble components in certain circumstances.
EFFECT: method enables reducing the time of concentrated sapropel production and improving the product quality.
1 dwg, 1 ex
SUBSTANCE: invention refers to medicine, namely to balneology. A balneological agent for treating and preventing various diseases is prepared by the gradual and sequential mixing at room temperature of yellow clay, natural brine of Bolshoy Tambukan Lake, sage essence, dimethyl sulphoxide in certain relations.
EFFECT: agent possesses the more prominent therapeutic effect.
5 tbl, 2 ex
SUBSTANCE: application of 5% water solution of sodium and potassium salts of humic acids, obtained from leonardite brown coal, in dose 10.0 mg/kg as membrane-protective preparation.
EFFECT: application of composition described above makes it possible to extend arsenal of membrane-protective preparations from low-toxic, cheap, available natural raw material.
2 dwg, 2 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: claimed invention relates to the field of medicine, namely to pharmacology, and describes a composition, containing fulvic acid or its salt and an antifungal compound, selected from fluconazole and amphotericin B. In accordance with the first version of the implementation the composition contains about 10 ml/kg of a solution of from about 0.25% to about 1% (wt/vol) fulvic acid or its salt and of about 10 mg/kg fluconazole. In accordance with the second version of the implementation the composition contains 0.25% (wt/vol) of the solution of fulvic acid or its salt and from about 0.06 mg/l to about 0.5 mg/l of amphotericin B.
EFFECT: invention can be used in a method of treating a fungal infection of the human or animal body, the method includes the introduction to an object, requiring it, of a therapeutically effective quantity of the composition.
11 cl, 1 dwg, 10 tbl, 3 ex
SUBSTANCE: method of obtaining a sapropel concentrate includes the extraction of easily separable water-soluble components of a native sapropel by the centrifugation and separation of a supernatant, with the supernatant being discharged into a vacuum evaporating aggregate, in which the concentration of the claimed supernatant is realised by the evaporation in vacuum, and the sapropel sediment is placed in a drying chamber, separated from the vacuum chamber with a net and a hermetic lock, heated with the simultaneous creation of a vacuum in the vacuum chamber, after which the sapropel sediment is subjected to a vacuum-impulse influence, the remaining moisture separated from the sediment is pumped by a vacuum pump into the vacuum evaporating aggregate, where it is condensed and concentrated at the same modes as the supernatant, after that, the dry sediment (extraction cake) is milled and packed, and the obtained concentrate is poured into vessels and each of the obtained target products is applied by purpose or the solubilisation of the milled dry sediment (extraction cake) is realised in the obtained concentrate of easily separable water-soluble components under certain conditions.
EFFECT: method makes it possible to preserve all useful substances of the native sapropel.
SUBSTANCE: peat in a mixture with water is loaded into a dispersion chamber, then the dispersion chamber is pressurised, static pressure of 5-7 atm is supplied into it and the content of the chamber is processed by ultrasonic vibrations with a density of insonification not less than 50 W/cm2, providing the sound pressure on the mixture of peat with water for the preliminarily specified time, which exceeds the static pressure by 2-3 times. The invention also relates to biogel, obtained by the claimed method.
EFFECT: obtaining water-peat biogel with nanosized particles, in which useful substances do not lose their efficiency.
SUBSTANCE: method for preparing an agent showing anti-inflammatory action involves an alkali treatment of terrestrial peat and extraction in the alcohol-chloroform mixture. The terrestrial peat preliminarily ground and dried is milled in the AGO planetary activator before the extraction in the presence of solid alkali and sodium percarbonate for 1 min.
EFFECT: method enables higher antioxidant and anti-inflammatory activity of the terrestrial peat agent by increasing a yield of the extractive biologically active substances.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to the pharmaceutical industry, namely to a medicated mud composition. The medicated mud composition contains prepared sludge argillaceous deposits of Tambukan Lake, Witepsol W 35, Witepsol H 15, polyethylene glycol 1500, Cremophore RH, as well as carbon-dioxide herbal extracts specified in a group consisting of at least the common willow, sea buckthorn, oak, calendula, licorice, brier, sage and/or carbon-dioxide extract of propolis taken in certain relations.
EFFECT: composition enables extending the range of medicinal agents using therapeutic sludge argillaceous deposits.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to the pharmaceutical industry, namely to a treating-preventive mud composition. The treating-preventive mud composition contains preliminarily prepared silt-clayey deposits of lake Tambukan, polyethyleneglycol 4000, polyethyleneglycol 1500, cremophor RH, as well as a carbon dioxide extract of propolis and/or a carbon dioxide vegetable extract, selected from the group, which includes, at least, eucalyptus, dog-rose, hop, laurel, chamomile, licorice and oak, taken in a specified ratio of components.
EFFECT: composition has an increased treating-preventive activity with respect to a patient's organism.
FIELD: medicine, phthisiology.
SUBSTANCE: in early period the preparation "Yantar-antitoks" is used in the dose 0.5 g, 3 times per a day for one month in addition to the conventional antituberculosis therapy. Invention promotes to elimination of symptoms associated with the total intoxication, resorption of focus-infiltration changes, elimination of destructions and ceasing secretion of microorganisms for shorter periods. Invention can be used in treatment of pulmonary tuberculosis.
EFFECT: improved method for treatment.
4 tbl, 1 ex