Method for preparing biological referent materials for producing material standards of compositions of these materials containing toxic metals and biological material produced by this method (versions)

FIELD: medicine.

SUBSTANCE: method involves preparing a biological referent material on the basis of a biological material of animal origin, after alimentary introduction to an animal of water-soluble lead or mercury or cadmium salts before ensuring in the biological materials of the certain lead or mercury or cadmium concentrations. Thereafter, the animals are killed by decapitation. Further, the biological material is sampled with an aliquote being analysed for the preset concentrations of toxic metals, then the biological material is packed and lyophilised.

EFFECT: invention allows improving diagnosing toxic metal poisonings and providing higher accuracy of determination of the toxic metal concentrations.

11 cl, 3 tbl, 3 ex

 

Group of inventions relates to toxicology, ecology, hygiene and veterinary medicine, namely to a method for biological reference materials for the production of standard samples of composition of these materials containing toxic metals, as well as to biological material obtained by the proposed method, and can be used in various fields to improve the diagnosis of poisoning by toxic metals and increase the accuracy of detection of toxic metals.

The invention uses the following adopted in this area terminology and abbreviations.

Nutritional food coming into the body with food through the gastrointestinal tract.

Biological media - individual organs, tissues, cells or subcellular structures of a living organism, having structural and functional homogeneity.

The biomaterial is part of organs, tissues, cells or subcellular structures of the body.

The decapitation - killing by cutting off their heads.

Standard sample (composition) - [term specified with regard to its definition, given in GOST 8.315-97, as well as in international legal instruments] sample of a substance or material, chemical composition or physical properties are:

typical for a specific group of substances or materials;

- identified need that is exactly what thew;

- have a high constancy;

- certified by the certificate.

Reference material "as material (substance) of the standard sample can be used reference material, i.e. a material (substance), sufficiently homogeneous and stable with respect to one or more specific properties used in accordance with intended use in a measurement process [R-SI. Samples of materials and substances standard. Terms and definitions; ISO VIM: 1993]".

Toxic - poisonous, causing negative changes in structures and/or functions of molecular, supramolecular, cell structure, tissues, organs and the organism as a whole.

Toxic metals (TM) is a conditional group metals, non-bearing biological functions in the body, but are prone to bioaccumulation, toxic even in small quantities [pollution of the Arctic 2002. Programme for monitoring and evaluation of the Arctic environment. AMAP. Oslo. - 2002. - 110 S.], which include lead, mercury, cadmium, arsenic, thallium, vanadium, Nickel, chromium, uranium, and sometimes other metals.

Lyophilization = freeze drying is a method of dehydration of biological material, lies in its freezing, followed by drying in a vacuum... [Encyclopedic dictionary of medical terms. Under. Ed. by Acad. B.V. Petrovsky Vol.2. M: the Soviet encycloped who I am. - 1983. 448 S.].

VL. century - wet weight, weight of native (natural) material.

dry. century - dry weight, weight of dried material.

Action TM (lead, mercury, cadmium, bismuth, etc.) on a living organism is in the center of attention of specialists of a number of scientific fields, namely, toxicologists, pathologists, veterinarians and ecologists. The relevance of this interest attaches to the ongoing industrialization of society and the growth of environmental contamination by these contaminants, the increase in the number of various incidents involving the release to the environment of the TM and the discovery of new aspects of the negative effects of TM on the body. The analysis of the content of TM metals in biological environments of the organism, the environment, food is an integral component of scientific, medical, ecological and sanitary-hygienic measures during the development of this problem.

When conducting a content analysis of TM using the following methods: atomic absorption spectrophotometry, Stripping voltammetry, flameless photometry, the so-called method of "cold vapor atomic emission spectrophotometry, mass spectrometry with inductively coupled plasma, etc.

It should be noted that virtually every one of the used methods involves the application of the relevant is standards, i.e. materials comparison containing the analyzed metal in certain amounts close to that in the samples and also close to the analyzed sample.

The application of the standard gives an idea about the adequacy of the method used (including sample preparation) and, ultimately, is the basis of quantitative characteristics of the studied component in the studied material. For the most part in work of this kind is used aqueous solutions of salts of the investigated metals, such as the State Standard Sample Solutions of Mercury (GSO PP), etc. This approach is justified if the study sample is an aqueous solution of the component or close to it.

For analysis of solid, viscous and other materials with a complex matrix (structure), such as BioTrade, soil, food, etc., it is necessary to apply the procedure of sample preparation, sometimes quite complex, with the purpose of the translation component in the liquid aqueous phase. Standards for some of such tasks are created and used for the analysis of relevant materials on the content of TM. For example, the standards of whole blood - [NIST-966 - Toxic metals in blood]; or standard [AMIB-1701 - toxic metals in human blood, 1 LGC. Setting standards in analytical science http://www.lgc.co.uk)]; or standards - grain rye SRI-01 (high content of Pb and Cd) - [GSO 8636-2004 (the "ECOLAN", http://ecolan.com.ru); GSO composition of soil - GSO 2498-2500-83].

In the analysis of such materials is a key issue, which ensures the reliability of the data, is the adequacy of the matrix standard and researched material, i.e. the maximum similarity in the structure of the standard and of the investigated material in which the desired component is also in the same conditions (chemical communications, environment, valence, ligands, etc.).

One of the most toxic of the group TM, included in the lists of priority pollutants a number of international organizations, including who and UNEP is the lead. High need households in this metal, the relative simplicity of its receipt and a large prevalence in the earth's crust (16 mg/kg) made him one of the first in a series of industrial poisons and global pollutants. Anthropogenic lead load exceeding geogenous 10-40 times, is able to induce violations at almost all levels of biological organization, including ecosystems [Lietuvninkas A.I. Anthropogenic geochemical anomalies and the natural environment. - Tomsk: Publishing house of NTL, 2002. - C.122-221].

It should be noted that the number of lead in biological media of the body, blood is the most important criterion of toxic action of the toxicant and the basis for the diagnosis [Livanov GA, With the Olev MB, Revich B.A. Lead hazard and population health: clinical features, diagnosis, and treatment toxicity. In Proc. of: Lead and children's health: diagnosis, treatment, prevention / edited Specimena. - SPb.: Publishing center "Caro", 1999. - P.18-26].

Known and used methods for the determination of lead in biological materials, foods and the environment - [MUK 4.11482-03 and pain 4.11483-03 "Determination of chemical elements in biological environments and preparations methods emission spectrometry with ionization-coupled plasma and mass spectrometry with ionization-coupled plasma"; Templeton, D.M. Qualifying cadmium and lead in whole blood // dim. Chem., v. 38, 1992. - P.1927-1929; Christensen J.M., J. Kristiansen // Lead, in Handbook on metals in clinical and analytical chemistry, Seiler H.G, A. Siegel, Siegel H (eds.) N.Y. Dekker. - 1994].

Another, extremely dangerous environmental pollutant is mercury (and its compounds). Industrial extraction of mercury increasing in recent decades, and its wide application in various spheres of human activity lead to an increase in its level in the environment, humans and animals, the development of serious conditions associated with excessive penetration and accumulation of mercury in the body. In this regard, there is an urgent need for the creation and development of methods for reliable determination of mercury in biological whom such materials, food, environmental objects.

Known methods for the determination of mercury in biological materials, foods and the environment [HOWTO. MUK 4.1.005 - 4.1.008 - 94, the state Committee of sanitary and Russia. - M., 1994, 29 S.]. For example, determination of mercury in soil and soil - [State Committee of the Russian Federation on the protection of the environment. LIM f 16.1:2.23-2000]; in food and the environment - [Rapoport E Use mercury analyzer type "Julia-2 for the analysis of food and environmental objects //Laboratory news of the Far East. The electronic version of the journal. No. 2-99. www.primer.ru/dvlab/dvlab_2/magazine.2htm]; in biological materials [Chomik LI, Kowalski YG, Overnight B.C. Determination of mercury in human biomaterials Laboratory news of the Far East. The electronic version of the journal. No. 2-99. www.primer.ru/dvlab/dvlab_2/magazine.2htm].

Cadmium also is recognized as one of the priority environmental pollutants. Cadmium, like all TM, lowers the overall resistance of the organism, its protective and adaptive capabilities weakens the immune system, violates the biochemical balance in the body. Before toxicologists and ecologists are the problem of objective evaluation and prediction of the degree of contamination of the environment by cadmium, as well as extensive work on the restriction of e is about revenue in the external and internal environment [Set J.E., Revich B.A., Yanin E.P. and other Geochemistry of the environment. M.: Nedra. 1990. - 333 S.; Limin BV, Mikulov VG, butchers ACTING, Patsyuk N.A., Rock AV, Carnacina T.S. Hygienic diagnostics of environmental pollution with heavy metals salts. SPb.: State medical Academy n.a. Mechnikov. 2003. - 123 S.].

Known methods for the determination of cadmium in biological materials and the environment - [Ishchenko A.A., Ganeeva A.A., Ivanenko NB etc. // Direct determination of cadmium in blood and plasma by atomic absorption with electrothermal atomization // proc. Dokl. at the II all-Russian Conf. "Analytical instruments". SPb., 2005, s-246; MUK 4.11482-03 and pain 4.11483-03 "Determination of chemical elements in biological environments and preparations methods emission spectrometry with ionization-coupled plasma and mass spectrometry with ionization-plasm"].

A method of obtaining biological materials for the production of standard samples of composition of these materials containing TM, adopted as a prototype method [http:/www.nist.gov/srm - SRM 966, National Institute of Standards & Technology, Certificate of Analysis, Standard Reference Material® 966, Toxic Metals in Bovine Blood], according to which the standard material is prepared on the basis of biological material - blood, obtained from cattle (bull), after alimentary injection of lead nitrate; collected krovjanistyh in the presence of TM.

Known use of biological materials in the process of diagnosis of intoxications TM [U.S. patent No. 5217594; U.S. patent No. 5368707; U.S. patent No. 5873990; U.S. patent No. 5468366].

The material for the standard referred to in U.S. patent No. 5468366, which is taken as a prototype biological reference material derived from biological raw materials, namely from bovine blood, and has a range of lead content 70±30 and 258±40 µg/DM3(depending on preparation conditions), has a high viscosity due to the high content of proteins and other blood components, and contains, as a preservative substance isothizolone (<0.002%). Liquid material is dispensed by the pipette. Storage conditions determine the shelf life of this control material from one week at room temperature, i.e. no higher than 24°C, up to a year frozen at -20°C.

Conducted information searches, as well as data obtained during experimental studies allow us to conclude that the creation of biological materials containing TM, and the development of a method of obtaining biological materials for the production of standard samples of composition of these materials containing TM is the actual problem.

Task to be solved by the present invention is directed, is to develop a method of obtaining biologically the reference materials for the production of reference materials of composition of these materials, containing TM.

The technical result, which is manifested in the use of the invention is obtaining a biological reference material containing specified concentrations of TM, for the production of standard samples of composition of these materials with the specified HM content.

To solve the problem and achieve the technical result of the proposed group of inventions, United by a common inventive concept.

One of the aspects of the proposed group of inventions is a method for biological reference materials for the production of standard samples of blood containing the toxic metal mercury, providing for the preparation of biological reference material on the basis of biological material obtained from animals after alimentary dosed introduction of water-soluble salts of toxic metal with subsequent analysis of the collected biological material in the presence of a toxic metal, the animals used in laboratory outbred rats, which gradually introduced a water-soluble salt of mercury to advances in biological material specified concentrations of mercury from 41 mg/DM3to 206 mg/DM3(VLV), then carry out the killing of laboratory outbred rats by decapita the AI and sampling of biological material in the form of blood, the sample which is subjected to analysis on a given concentration of mercury, after which the biological material is Packed and lyophilizers.

Another aspect of the invention is a biological reference material for production of normal blood samples containing toxic metal mercury, obtained as described above and representing a lyophilized blood mercury levels from 256 μg/kg to 1287.5 µg/kg (suhu)intended for creating WITH blood containing mercury.

Another aspect of the present invention relates to a method for biological reference materials for the production of standard samples of the kidney containing the toxic metal mercury, providing for the preparation of reference material on the basis of biological material obtained from animals after alimentary dosed introduction of water-soluble salts of toxic metal with subsequent analysis of the collected biological material in the presence of a toxic metal, the animals used in laboratory outbred rats, which gradually introduced a water-soluble salt of mercury to advances in biological material mercury concentrations from 7.72 mg/kg and 264 mg/kg (VLV), then carry out the killing of outbred laboratory rats by decapitate and for the PRS biological material in the form of tissues or organs, the sample which is subjected to analysis on a given concentration of mercury, after which the biological material is Packed and lyophilizers.

Another aspect of the present invention relates to a biological reference material for the production of standard samples of the kidney containing the toxic metal mercury, obtained as described above from the kidneys and representing a lyophilized buds with mercury in it from 1020 µg/kg up to 1320 mg/kg (suhu)used to create FROM the kidneys containing mercury.

Another aspect of the present invention relates to a biological reference material for the production of standard samples of brain tissue containing the toxic metal mercury, obtained as described above from brain tissue and represents a lyophilized brain tissue content of mercury from 15.4 µg/kg up to 103.6 mg/kg (suhu)used to create WITH brain tissue containing mercury.

Another aspect of the present invention relates to a biological reference material for the production of standard samples of muscle tissue containing the toxic metal mercury, obtained as described above from the muscle tissue and represents a lyophilized muscle tissue with mercury from 41.2 µg/kg to 176.6 mg/kg (suhu), designed on what I create WITH muscle tissue, containing mercury.

Another aspect of the present invention relates to a biological reference material for the production of standard samples of adipose tissue containing the toxic metal mercury, obtained as described above from adipose tissue and represents a lyophilized adipose tissue content of mercury from 25.2 μg/kg to 118.8 mg/kg (suhu)used to create WITH adipose tissue containing mercury.

Another aspect of the present invention relates to a biological reference material for the production of standard samples of liver tissue containing the toxic metal mercury, obtained as described above from liver tissue and represents a lyophilized liver content of mercury from 122 mg/kg up to 651 mg/kg (suhu)used to create WITH liver tissue containing mercury.

Another aspect of the present invention also relates to a biological reference material for the production of standard samples of bone tissue containing the toxic metal mercury, obtained as described above from the bone and representing a lyophilized bone tissue mercury from 30.9 μg/kg to 137.6 mg/kg (suhu)used to create WITH bone tissue that contains mercury.

Another aspect of the present invention refers to the manner in which Holocene biological reference materials for the production of standard samples of blood, containing the toxic metal cadmium, providing for the preparation of reference material on the basis of biological material obtained from animals after alimentary dosed introduction of water-soluble salts of toxic metals with subsequent analysis of the collected biological material in the presence of toxic metals, at the same time as animals use laboratory outbred rats, which gradually introduced a water-soluble salt of cadmium to advances in biological material concentrations of cadmium from 39 ág/DM3up to 64 µg/DM3(VLV), then carry out the killing of laboratory outbred rats by decapitate and sampling of biological material in the form of blood, the sample which is subjected to analysis on a given cadmium, after which the biological material is Packed and lyophilizers.

Another aspect of the present invention relates to a biological reference material for production of normal blood samples containing toxic metal cadmium, obtained as described above and representing a lyophilized blood cadmium from 243.8 ág/kg 400 ág/kg (suhu)intended for creating WITH blood containing mercury.

Thus, the proposed group of inventions relates to the same object in the Yes, the same purpose, providing the same technical result.

The possibility of objective manifestations of the technical effect of the invention is confirmed by reliable data given in the examples containing information, experimental, obtained according to the methods adopted in this field.

The invention is illustrated by tables.

Table 1 presents data showing the effect of dose of dose of mercury on the contents of this TM in the blood of experimental animals.

Table 2 presents data on the effect of dose of dose of mercury on the contents of this toxicant in some organs and tissues of rats.

Table 3 presents data showing the effect of dose of dose of cadmium on the contents of this TM in the blood of experimental animals.

The invention is illustrated in the examples of the method of obtaining the biological reference materials for the production of standard samples of composition of these materials containing TM, namely:

example 1 - obtaining a biological reference material for the production of standard samples of blood containing mercury;

example 2 - obtaining a biological reference material for the production of reference materials of composition of kidney, brain tissue, isichei fabric, adipose tissue, liver tissue and bone tissue containing mercury;

example 3 - obtaining a biological reference material for the production of standard samples of blood containing cadmium.

Example 1. Obtaining a biological reference material for the production of standard samples of blood containing mercury.

Before the experiment was carried out by training animals to experiment - keeping for two weeks in quarantine for adaptation and determine daily water consumption, weighing. Possible salts of toxic metals selected water-soluble, giving acceptable in the experiment lasting solutions - as a rule, the acetates and nitrates, qualifications not lower than "analytical grade". In the research was used nitrate of mercury - Hg(NO3)2.

The experiments were conducted in a laboratory outbred rats weighing 180 to 200 g, which in drinkers was added the calculated amount of a solution of salts of mercury, which they consumed with drinking water and the number of which was determined on the basis of pre-existing data about the consumption of animals.

For preparation of this solution was used nitrate of mercury, M.V. 324.6, analytical grade. After the calculation was carried out based on the weight of the cation of mercury (Hg+2)preparing an aqueous solution n the waste mercury adding in which drinkers four groups of animals taking into account body weight and water consumption of rats were provided admission to their bodies cation of mercury, respectively: 0.06 mg/kg (group 1); 0.3 mg/kg (group 2); 0.6 mg/kg (group 3) and 1.8 mg/kg (group 4) per day. The seed of animals was carried out as follows. In drinkers (glass bottles of 0.5 DM3for the group of rats daily for 30 days in a row was added to the solution of nitrate of mercury in the volume, which was calculated by the equation: X=(m×d×0.45)/c×v, where:

X is the number of solution of mercury (DM3)deposited in the drinking volume 0.450 DM3;

m is the mass per animal (kg) (group of animals - average);

d - daily dose of metal cation (mg/kg);

v is the average daily consumption per animal (DM3), calculated on the basis of water consumption in all rats of the group on the previous day;

with the concentration of the metal cation (mg/DM3) in solution;

0.45 - end solution volume (DM3).

After making the solution of nitrate of mercury in drinkers poured tap water up to the mark 0.45 DM3. In this case, the consumption of water with the addition of a specified amount of a solution of nitrate of mercury should ensure that the intake of each rat per day on average 0.06 mg/kg, 0.3 mg/kg, 0.6 mg/kg and 1.8 mg/kg Hg+2.

At the end of this period alive is the shaft was cut and collected the biological material.

The calculated daily dose of mercury and actually received the amount of mercury in the course of the experiment are presented in table 1. The data presented in table 1 demonstrate the effect of dose priming on HM content in the blood of experimental animals. During the experiment there were used the following methods of analysis: atomic absorption spectroscopy method cold vapor; atomic absorption spectroscopy method cold vapor with Zeeman correction; atomic absorption spectroscopy with electrothermal atomization.

Table 1
The specified dose for weed, mg/kgExperimentally injected dose, mg/kgThe content of Hg2+in whole blood groups of rats, mg/DM3(VLV)
Methods for the determination of mercury
No.Atomic absorption spectroscopy method cold vaporAtomic absorption spectroscopy method cold vapor with Zeeman correctionAtomic absorption
spectroscopy with electrothermal atomization
groups of rats
on Hg2+on Hg2+
11.81.82±0.04-206 (1287.5)194 (1212.5)
20.60.61±0.0196 (600)96 (600)120 (759)
30.30.29±0.00560 (375)75 (469)62 (388)
40.060.06±0.00144 (275)59 (369) 41 (256)
Note: In parentheses are the values of the mercury content in the appropriate biological reference material on dry weight of the latter (suhu), i.e. in mg/kg

From the data presented in table 1, there is a clear dependence of the content of fluorine in the blood of experimental animal dose of toxicant, as well as observed the practical coincidence data obtained by three different methods.

Vials packaged blood corked with rubber stoppers and placed in a low temperature refrigerator (t<-15°C) not less than 10 hours. Vials of frozen blood is connected to vacuum drying to liophilization "Freeze Dryer 3" firm Labconco (USA) for 8 hours at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight. During this time there is vacuum-drying material, the blood turns into a red-brown crystalline mass, which is easily destroyed in the powder with a light tap on the bottle.

Thus, the above described biological reference material is a brown crystalline powder organ-mineral matrix (freeze-dried) blood mercury levels from 256 μg/kg to 1287.5 µg/kg (suhu).

Example 2. Obtaining a biological reference material for production is as standard samples of composition of kidney, brain tissue, muscle tissue, adipose tissue, liver tissue and bone tissue containing mercury.

Before the experiment was carried out by training animals to experiment - keeping for two weeks in quarantine for adaptation and determine daily water consumption, weighing. Possible salts of toxic metals selected water-soluble, giving acceptable in the experiment, lasting solutions, usually chlorides, acetates and nitrates, qualifications not lower than "analytical grade". The surveys were used acetate mercury - Hg(CH3COO)2.

Experiments conducted on laboratory outbred rats weighing 180 to 200 g, which in drinkers was added the calculated amount of a solution of acetate of mercury, which they consumed with drinking water and the number of which was determined on the basis of pre-existing data about the consumption of animals.

For preparation of this solution was used acetate mercury, M.V. 318.7, analytical grade. After the calculation was carried out based on the weight of the cation of mercury (Hg+2)preparing an aqueous solution of acetate of mercury, adding in which drinkers three experimental groups of animals taking into account body weight and water consumption of rats were provided admission to their bodies cation of mercury, respectively: 0.06 mg/kg (group 1); 0.32 mgkg (group 2); 0.65 mg/kg (group 3) per day.

Methods of calculation of the dose of a toxicant to obtain a given concentration of mercury in blood were calculated by the equation: X=(m×d×0.45)/c×v given in examples 1 and 2.

Method of atomic absorption spectrometry cold vapor was determined by the content of mercury in biological materials - kidney, brain tissue, muscle tissue, adipose tissue, liver tissue and bone tissue and the effect of dose of dose of mercury on the content of fluorine in biological reference material (tissues and organs of the rat). The data presented in table 2.

Table 2
The mercury content in the native material, µg/kg (VLV)
Dose
mercury
(mg/kg)KidneyMuscle tissueAdipose tissueLiver tissueBone
204
0.06(1020)7.72 (15.4)20.6 (41.2)12.6 (25.2)24.4 (122)26.3 (30.9)
249
0.32(1245)24.5 (49)39.9 (79.8)23.8 (47.6)104 (520)53.1 (62.5)
264
0.65 (1320)51.8 (103.6)88.3 (176.6)59.4 (118.8)130.2 (651)117 (137.6)
Note: In parentheses are the values of the mercury content in the corresponding reference biological material to dry in the last suhv), i.e. in mg/kg

At the end of this period, animals kill, excised biological material - buds and 2 g of this body is placed in a glass vial and then the vial with biological material in the native form is subjected to freezing for at least 10 hours at a temperature of not higher than -15°C and then hold lyophilization at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight.

The result is dried organo-mineral matrix (lyophilisate) kidney with a specified content of mercury from 1020 µg/kg up to 1320 mg/kg (suhu).

At the end of this period, animals kill, excised biological material brain, then 2 g of brain tissue is placed in a glass vial and then the vial with biological material in the native form is subjected to freezing for at least 10 hours at a temperature of not higher than -15°C and then hold lyophilization at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight.

The result of organo-mineral matrix (lyophilisate) of brain tissue with a given content of mercury from 15.4 µg/kg up to 103.6 mg/kg (suhu).

At the end of this period, animals kill, excised biological material - muscle, then 2 g of skeletal muscle is placed in a glass vial and then the vial with biological material in the native form is subjected to freezing for at least 10 hours at a temperature of not higher than -15°C and then hold lyophilization at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight.

The result of organo-mineral matrix (lyophilisate) muscle tissue with the specified content of mercury from 41.2 µg/kg to 176.6 mg/kg (suhu).

At the end of this period, animals kill, excised biological material adipose tissue of the mesentery, and then 2 g of adipose tissue of the mesentery placed in a glass vial and then the vial with biological material in the native form is subjected to freezing for at least 10 hours at a temperature of not higher than -15°C and then hold lyophilization at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight.

The result of organo-mineral matrix (lyophilisate) adipose tissue with a given content of mercury from 25.2 MK is/kg to 118.8 mg/kg (suhu).

At the end of this period, animals kill, excised biological material - the liver, then 2 g of liver was placed in a glass vial and then the vial with biological material in the native form is subjected to freezing for at least 10 hours at a temperature of not higher than -15°C and then hold lyophilization at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight.

The result of organo-mineral matrix (freeze-dried) liver tissue with a given content of mercury from 25.2 μg/kg to 118.8 mg/kg (suhu).

At the end of this period, animals kill, excised biological material - the bones of the skeleton, and then 2 g of the bones of the skeleton placed in a glass vial and then the vial with biological material in the native form is subjected to freezing for at least 10 hours at a temperature of not higher than -15°C and then hold lyophilization at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C to constant weight.

The result of organo-mineral matrix (lyophilisate) of bone tissue with a given content of mercury from 30.9 μg/kg to 137.6 mg/kg (suhu).

Example 3. Obtaining a biological reference material for the production of standard samples of blood containing cadmium.

Before the experiment was carried out by preparing a stomach who's to experiment keeping within two weeks in quarantine for adaptation and determine daily water consumption, weighing. Possible salts of toxic metals selected water-soluble, giving acceptable in the experiment lasting solutions - as a rule, nitrates and acetates, qualifications not lower than "analytical grade". In the research was used cadmium nitrate Cd(NO3)2.

Experiments conducted on laboratory outbred rats weighing 180-200 g, in which drinkers was added the calculated amount of a solution of cadmium nitrate, which they consumed with drinking water and the amount of which was determined on the basis of pre-existing data about the consumption of animals.

For preparation of this solution was used cadmium nitrate (neutral) Cd(NO3)2·4H2O, M.V. 309.47, qualification "analytical grade".

After the calculation was carried out based on the molecular weight of the cation cadmium (112.4), was prepared aqueous working solution of cadmium nitrate, adding in which waterers animals taking into account body weight and water consumption of rats were provided admission to their bodies cation cadmium 0.06 mg/kg and 0.031 mg/kg per day.

The seed of animals was carried out as follows. In drinkers (glass bottles of 0.5 DM3for the group of rats daily for 30 days is successively added to the solution of lead acetate in the amount which was calculated by the equation:

X=(m×d×0.45)/c×v given in example 2.

After making the solution of cadmium nitrate in drinking poured tap water up to the mark 0.45 DM3. In this case, the consumption of water with the addition of the specified amount of the basic solution of cadmium nitrate should ensure that the intake of each rat per day required number of Cd+2(see table 3).

The content of cadmium in the blood of rats was determined by atomic absorption spectrometry with electrothermal atomization (see table 3).

Table 3
The content of the Cd2+in the blood
Dose Cd2+,Method for determining
mg/kgcadmium
ág/DM3(VLV)µg/kg (suhu)
0.0639243.8Atomic absorption is
spectroscopy with electro-
0.3164400thermal atomization

Upon reaching the preselected concentration of cadmium in the blood of rats were killed by decapitate and collected blood clotting. Blood was Packed up vials - 2 g, froze when <-15°C.

Vials of frozen blood is connected to vacuum drying to liophilization "Freeze Dryer 3" firm Labconco (USA) at a pressure of less than 10-4mm RT. Art. and temperatures below -50°C for 8 hours. During this time there is vacuum-drying material, the blood turns into a red-brown crystalline mass, which is easily destroyed in the powder with a light tap on the bottle.

Thus, the above described biological reference material is a brown crystalline powder organo-mineral matrix (freeze-dried) blood cadmium from 243.8 ág/kg 400 ág/kg (suhu).

1. A method of obtaining a biological reference materials for the production of standard samples of blood containing the toxic metal mercury, providing for the preparation of biological reference material based bio is oricheskogo material, obtained from animals after alimentary dosed introduction of water-soluble salts of toxic metal, with subsequent analysis of the collected biological material in the presence of a toxic metal, characterized in that as animals use laboratory outbred rats, which gradually introduced a water-soluble salt of mercury to achieve in a biological material selected concentration of mercury from 41 to 206 mg/DM3(VLV), then carry out the killing of laboratory outbred rats by decapitate and sampling of biological material in the form of blood, the sample which is subjected to analysis on a given concentration of mercury, after which the biological material is Packed and lyophilizers.

2. Biological reference material for production of normal blood samples containing toxic metal mercury, characterized in that it is obtained by the method according to claim 1 and is a lyophilized blood mercury levels from 256 to 1287,5 µg/kg (suhu), designed to create a standard blood sample containing mercury.

3. A method of obtaining a biological reference materials for the production of standard samples of the kidney containing the toxic metal mercury, providing for the preparation of reference material on the basis of the biological material is a, obtained from animals after alimentary dosed introduction of water-soluble salts of toxic metal, with subsequent analysis of the collected biological material in the presence of a toxic metal, characterized in that as animals use laboratory outbred rats, which gradually introduced a water-soluble salt of mercury to advances in biological material mercury concentrations from 7,72 to 264 mg/kg (VLV), then carry out the killing of laboratory outbred rats by decapitate and sampling of biological material in the form of tissues or organs of the sample which is subjected to analysis on a given concentration of mercury, after which the biological material is Packed and lyophilizers.

4. Biological reference material for the production of standard samples of the kidney containing the toxic metal mercury, characterized in that it is obtained by the method according to claim 3 of the kidney and is a lyophilized buds with mercury in it from 1020 to 1320 mg/kg (suhu), designed to create a standard sample of the kidney containing mercury.

5. Biological reference material for the production of standard samples of brain tissue containing the toxic metal mercury, characterized in that it is obtained by the method according to claim 3 of the brain tissue and is the Wallpaper of the freeze-dried brain tissue content of mercury from 15,4 to 103,6 µg/kg (suhu), designed to create a standard sample of brain tissue containing mercury.

6. Biological reference material for the production of standard samples of muscle tissue containing the toxic metal mercury, characterized in that it is obtained by the method according to claim 3 of the muscle tissue and is a lyophilized muscle tissue with mercury from 41.2 per cent to 176,6 µg/kg (suhu), designed to create a standard sample of muscle tissue containing mercury.

7. Biological reference material for the production of standard samples of adipose tissue containing the toxic metal mercury, characterized in that it is obtained by the method according to claim 3 of the adipose tissue and is a lyophilized adipose tissue content of mercury from 25,2 up of 118.8 mg/kg (suhu), designed to create a standard sample of adipose tissue containing mercury.

8. Biological reference material for the production of standard samples of liver tissue containing the toxic metal mercury, characterized in that it is obtained by the method according to claim 3 of the liver tissue and is a lyophilized liver content of mercury from 122 to 651 mg/kg (suhu), designed to create a standard sample of liver tissue containing mercury.

9. Biological reference material for the production of standard samples of coast is the second fabric, containing the toxic metal mercury, characterized in that it is obtained by the method according to claim 3 of the bone tissue and is a lyophilized bone tissue mercury from 30,9 to 137.6 mg/kg (suhu), designed to create a standard sample of bone tissue that contains mercury.

10. A method of obtaining a biological reference materials for the production of standard samples of blood containing the toxic metal cadmium, providing for the preparation of reference material on the basis of biological material obtained from animals after alimentary dosed introduction of water-soluble salts of toxic metals, with subsequent analysis of the collected biological material in the presence of toxic metals, characterized in that as animals use laboratory outbred rats, which gradually introduced a water-soluble salt of cadmium to advances in biological material concentrations of cadmium from 39 to 64 µg/DM3(VLV), then carry out the killing of laboratory outbred rats by decapitate and sampling of biological material in the form of blood, the sample which is subjected to analysis on a given cadmium, after which the biological material is Packed and lyophilizers.

11. Biological reference material for the production of the conventional blood samples containing the toxic metal cadmium, characterized in that it is obtained by the method according to claim 10 and is a lyophilized blood cadmium from 243,8 to 400 µg/kg (suhu), designed to create a standard blood sample containing cadmium.



 

Same patents:

FIELD: medicine.

SUBSTANCE: method includes increase of biomass of human germ stem cells of non-feeder line hESKM-05 with application of main medium mTeSR in flasks covered with matrigel. After that, performed is subcultivation by means of 0.05% dispase solution. Then, in order to form germ stem cells of embrioid bodies, by means of dispase cells are transferred into "коДМЕМ" medium with addition of 10% substituent of SR serum, 100 mcg/ml of sulfate canamicine, 1mM of L-glutamine solution and 1 mM solution of essential amino acids, as well as 2 mcg/ml solution of 5-asa-2-seoxycitidine or 2 mcM of sodium butirate solution and stimulated for 3 days. After that retransmission onto prepared collagen-chitosane matrix and cultivation in nutritional medium "коДМЕМ" with addition of 1 mM solution of essential amino acids, 1 mM of L-glutamine, 10% substituent of SR serum, 10-7 M of retinoic acid and 10 ng/ml of ascorbic acid are performed with replacement of solution every three days.

EFFECT: invention makes it possible to obtain cardiomyicytal matrix suitable for direct transplantation.

7 dwg, 1 ex

FIELD: medicine.

SUBSTANCE: stem cells of an adult human recovered from subcutaneous fat eye tissues are induced to differentiate in insulin-secreting cells in a medium containing cytokines and growth factors, including the B27 additive, fibroblast-2 growth factor, epidermal growth factor, nicotinamide, glucagon-like peptide-1, active A, insulin-like growth factor and betacelluline. For a cultivation process, high-concentrated glucose is replaced for low-concentrated glucose.

EFFECT: invention enables producing cells synthesising considerable amounts of insulin and C-peptide.

3 cl, 8 dwg, 2 tbl, 9 ex

FIELD: medicine.

SUBSTANCE: invention represents biotransplant for correction of soft tissue defects, characterised by the fact that it represents suspension which contains autologic culture of fibroblasts in 0.9% solution of sodium chloride in concentration 0.6-3.0×106 in 1 ml of biotransplant, integrated on pharmaceutically acceptable biocompatible biodegradable crushed to size of 100-200 mcm cell-free matrix in solution of pharmaceutically acceptable biocompatible biodegradable preparation of hyaluronic acid, volume ratio of autologic culture of fibroblasts, cell-free matrix and preparation of hyaluronic acid constitutes 4:1:1 respectively, and as pharmaceutically acceptable biocompatible biodegradable cell-free matrix used is preparation "Saimetra", representing processed donor human skin, deprived of cells and structural immunospecific proteins, whose basis consists of collagen and elastin, represented as injection form.

EFFECT: considerable reduction of quantity of injected cells, increase of their viability and ensuring long preservation of implant in affected tissue.

19 cl, 4 ex, 5 dwg

FIELD: medicine.

SUBSTANCE: substance of the invention involves a stem cell (SC) preparation with reprogrammed cell signalling containing a base SC preparation in a membrane and/or a nucleus and/or a cytoplasm of which there is implanted a protein or pharmaceutical preparation capable to regulate signalling pathways of the SC and nidus cells in a mammal's body, pre-encapsulated in a nanocontainer with dimensions of 100 nm, produced of a biodegradable material, intact for organelles and compartments of the SC of the base preparation. This material has the preset biodegradation time in a mammal's body to provide the programmed protein or pharmaceutical preparation recovery in an intra- or intercellular space and to implement thereby reprogramming of signalling transduction of the SC key genes in a required therapeutic pattern of physiological cell cycle events directly in a pathological body region or tissue.

EFFECT: provided target addition of the signalling substances strictly in the involved body region.

14 cl, 6 ex, 3 dwg

FIELD: medicine.

SUBSTANCE: method provides phagocyte recovery from a cell mixture. The recovered cells are mixed with medium 199 in 18 bottles, and the cells are attached to glass at room temperature for 60 minutes. In 9 bottles, a culture medium containing medium 199, L-glutamine and mixed human serum heated for 30 min at 52°C is added to the cells in the preset amounts to produce cell samples. A culture medium of said composition containing 0.01% of zymosan particles is added to another 9 bottles with the cells to produce the cell samples. The prepared cell samples are divided into three portions. One one-third potion of zymosan and zymosan-free samples are frozen. The second portion of the zymosan and zymosan-free cell samples are cultivated at temperature 37°C for 4 to 6 h to be frozen, and the third portion of the zymosan and zymosan-free samples are cultivated at temperature 37°C for 18 to 24 h to be frozen respectively. All 18 bottles are exposed to simultaneous multiple freezing to -10°C and thawing at room temperature to complete recovery of intracellular lysozyme with determination of amount by a micromethod and calculation of synthesised lysozyme and a phagocyte activation value (PAV) by formula: PAV=Z Lsynth - Z-free Lsynth, mcg/ml, where Z Lsynth is a difference of lysozyme amount in the cultivated zymosan cells and lysozyme amount in the uncultivated zymosan cells, mcg/ml; Z-free Lsynth is a difference of lysozyme amount in the cultivated zymosan-free cells and lysozyme amount in the uncultivated zymosan-free cells, mcg/ml; the phagocyte activation value for long-term activation mechanism assessment is calculated by said formula by results of cell cultivation for 4 to 6 hours, and the phagocyte activation value for long-term activation mechanism assessment is calculated by said formula by results of cell cultivation for 18 to 24 hours.

EFFECT: invention allows more precise phagocyte activation assessment.

4 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: there is conducted addition of biologically active substances (BAS) to a perfluorocarbon (PFC) emulsion containing at least: perfluordecahydronaphthalene and/or perfluormethylcyclohexylpiperidine and/or perfluortributylamine and/or perfluoroctylbromide stabilised by a proxanol solution and/or phospholipids. The PFC emulsion with the BAS is added to stem cells (SC) for co-cultivation. After cultivation, the SC are introduced to a subject requiring the BAS introduction.

EFFECT: invention allows providing targeted transport of the BAS into the body.

16 cl, 7 dwg, 1 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: method includes cultivation of a cell mixture containing cardiomyocytes and non-cardiomyocytes produced of stem cells, with the exception of human embryo cells, in a culture medium. A cell mixture is cultivated in the following conditions: (i) low glucose feed, (ii) one or more conditions selected from a group consisting of: the low calcium content; the low nutrient content; lactic acid feed; asparaginic/glutamic acid feed and pyruvic acid feed.

EFFECT: invention provides high purity and yield of cardiomyocytes from the cell mixture containing cardiomyocytes and non-cardiomyocytes produced of stem cells, with the exception of human embryo cells.

11 cl, 23 dwg, 1 tbl, 15 ex

FIELD: medicine.

SUBSTANCE: method includes cultivation of a cell mixture containing cardiomyocytes and non-cardiomyocytes produced of stem cells, with the exception of human embryo cells, in a culture medium. A cell mixture is cultivated in the following conditions: (i) low glucose feed, (ii) one or more conditions selected from a group consisting of: the low calcium content; the low nutrient content; lactic acid feed; asparaginic/glutamic acid feed and pyruvic acid feed.

EFFECT: invention provides high purity and yield of cardiomyocytes from the cell mixture containing cardiomyocytes and non-cardiomyocytes produced of stem cells, with the exception of human embryo cells.

11 cl, 23 dwg, 1 tbl, 15 ex

FIELD: medicine.

SUBSTANCE: method includes cultivation of a cell mixture containing cardiomyocytes and non-cardiomyocytes produced of stem cells, with the exception of human embryo cells, in a culture medium. A cell mixture is cultivated in the following conditions: (i) low glucose feed, (ii) one or more conditions selected from a group consisting of: the low calcium content; the low nutrient content; lactic acid feed; asparaginic/glutamic acid feed and pyruvic acid feed.

EFFECT: invention provides high purity and yield of cardiomyocytes from the cell mixture containing cardiomyocytes and non-cardiomyocytes produced of stem cells, with the exception of human embryo cells.

11 cl, 23 dwg, 1 tbl, 15 ex

FIELD: medicine.

SUBSTANCE: method involves preparing a bone marrow (BM) aspirate after a iliac bone puncture, analysing a BM sample for the presence of infectious agents and bacterial contaminations. Then the BM sample is placed in heparin water 15 ml produced by dilution of heparin 5 ml in physiologic saline 200 ml. A growth medium is prepared by adding fetal bovine serum 5 ml to the concentration not less than 10% preheated to temperature 37°C to the MEM-cc or Advanced Stem Cell Media medium 45 ml. After dilution of the BM sample in Hanks's solution in 1.5-2 times, the BM suspension is layered on ficoll in the ratio 30 ml of the suspension to 15 ml of ficoll, and the solution is centrifuged with acceleration 800G for 30 min; 2 upper layers are collected with lower of these layers shaped as a white ring with pure leukocyte isolation. These two layers are transferred to a test tube containing the MEM-wasps or Advanced Stem Cell Media culture medium 7 ml, and the produced solution is centrifuged with acceleration 400G for 5 min; a growth medium 5-10 ml is added to a precipitate, and the prepared suspension is pipetted on 1-2 bottles that is followed with stem cell cultivation.

EFFECT: invention allows higher uniformity and quality of producing the higher-purity MSC population.

FIELD: organic chemistry, natural compounds, medicine, oncology.

SUBSTANCE: invention represents new saponin mixtures used for inhibition of initiation and activation of mammalian epithelial cell in pre-malignant or malignant state, for stimulation of apoptosis of mammalian malignant cell, prophylaxis of anomalous proliferation of mammalian epithelial cell, for treatment of inflammatory and regulation of angiogenesis in mammal. These mixtures are isolated form plants of species Acacia victoriae. Also, invention relates to methods for their applying. These compounds can comprise triterpene component, such as acacic or oleanolic acid to which oligosaccharides and monoterpenoid components are joined. Mixtures and compounds elicit properties associated with regulation of apoptosis and cytotoxicity of cells and strong anti-tumor effect with respect to different tumor cells.

EFFECT: valuable medicinal properties of compositions.

43 cl, 53 tbl, 50 dwg, 44 ex

FIELD: biotechnology, molecular biology, medicine, genetic engineering, pharmacy.

SUBSTANCE: the hemopoietic protein comprises the amino acid sequence of the formula: R1-L1-R1, R2-L1-R1, R1-R2 or R2-R1 wherein R1 represents the modified ligand flt-3; R2 represents the modified human IL-3, the modified or unmodified colony-stimulating factor. Modification of R1 is carried out by addition of N-end with C-end directly or through linker (L2) that is able to join N-end with C-end to form new C- and N-ends. The modified human IL-3 is prepared by replacing amino acids at positions 17-123. The human G-CSF is modified by exchange of amino acids. The hemopoietic protein is prepared by culturing cells transformed with vector comprising DNA that encodes the hemopoietic protein. The hemopoietic protein stimulates producing hemopoietic cells and this protein is used as a component of pharmaceutical composition used in treatment of humans suffering with tumor, infectious or autoimmune disease. Invention provides preparing multifunctional hemopoietic proteins eliciting the enhanced activity with respect to stimulation of hemopoietic cells and eliciting the improved physical indices. Invention can be used for preparing chimeric multifunctional hemopoietic proteins.

EFFECT: improved preparing and producing method, valuable medicinal properties of protein.

22 cl, 19 dwg, 18 tbl, 117 ex

FIELD: cellular biology, medicine.

SUBSTANCE: invention relates to isolating and cryopreserving precursor-cells. Methods involve treatment of human liver tissue for preparing the essentially monocellular suspension containing precursor-cells and cells that are not precursor-cells, a single or more lines of cellular differentiation presenting in the human liver. Invention describes methods involving stage for separating cellular population resulting to reducing amount of cells that are not precursor-cells and providing preparing the separated suspension enriched with precursor-cells expressing one or more markers and associated with a single or more lines of the cellular differentiation. Also, invention describes a method for selection cells from the separated suspension wherein these cells or their progeny, or their more matured forms express one or more markers associated with lines of the cellular differentiation. These markers involve: CD14, CD34, CD38, CD45 and ICAM. Hepatic precursor-cells have diameter size 6-16 mc, they are diploid and show indices: glycoforin A-, CD45-, AFP+++, ALB+, ICAM+ and they comprise subpopulations varying with respect to expression of CD14+, CD34++, CD38++ and CD117++. These cells are useful for carrying out cellular and genetic therapy in liver treatment and for preparing artificial organs also.

EFFECT: valuable biological and medicinal properties of cells.

41 cl, 7 tbl, 13 dwg, 15 ex

FIELD: medicine, genetic engineering.

SUBSTANCE: invention relates to applying genetic engineering approaches for treatment of autoimmune diseases, in particular, for treatment of cerebrospinal sclerosis. This is achieved by incorporation of one or some recombinant genes encoding autoantigens that represent a target for autoimmune response. In particular, invention claims a method for designation of gene encoding encephalitogenous epitope of proteolipid protein and expression of gene product in vivo by using the recombinant retroviral vector. Expression and secretion of encephalitogenous epitope improves histopathological and clinical indices in experimental autoimmune encephalomyelitis in mice that is used as a model of cerebrospinal sclerosis. The advantage of invention involves the development of a method for recovery the tolerance in treatment of cerebrospinal sclerosis being without suppression of immune system.

EFFECT: improved and valuable method for treatment.

6 cl, 13 dwg, 3 ex

FIELD: biotechnology and pharmaceutical industry.

SUBSTANCE: title operations are accomplished by following way. Three-dimensional culture of stromal cells is cultured in piston flow bioreactor, in particular being introduced in fibrous matrix incorporated into substrate, which is placed in container constituting a part of bioreactor piston. Stromal cells are grown until density 5 x 106 cell/cm3 substrate is attained, whereupon non-differentiated hemopoietic cells are either sowed directly into piston flow bioreactor or cultured in conditioned medium of stromal cells obtained by gathering medium from indicated flow bioreactor. Non-differentiated hemopoietic cells obtained by cultivation in presence of three-dimensional culture of stromal cells or their conditioned medium are transplanted to into recipient.

EFFECT: enabled growth of large amounts of stromal cells within a relatively small volume to provide longer maintenance of vital activity and reproduction of non-differentiated hemopoietic stem cells or precursor cells.

77 cl, 9 dwg, 3 tbl

FIELD: veterinary science.

SUBSTANCE: the present innovation deals with serological diagnostics of infectious diseases in cattle, viral diarrhea as mucosal disease in cattle (cattle VD) and evaluation of immunity strength in vaccinated animals. The method deals with growing finite cell line of coronary vessels in cow's embryo in Eagle's MEM nutritive medium with 20 mg/ml kanamycin and 10% equine serum. Double dilutions of inactivated tested serumal samples (56 C, 30 min) should be mixed at equal volume (0.05 ml) with 100 TCD50/0.1 ml virus to be incubated for 1 h at 37 C in CO2-incubator. On finishing the incubation one should add 0.1 ml suspension of the above-mentioned finite cell line in Eagle's MEM nutritive medium containing 20 mg/ml kanamycin and 5% (2.5% final concentration) of equine serum. Cell concentration in the culture corresponds to 3.5 x 105. Reaction registration should be carried out in 3 d while cytopathogenic viral action appears in control holes containing working viral dosage. The innovation enables to widen the number of diagnostic immunological methods for veterinary purposes.

EFFECT: higher efficiency of detection.

1 cl, 7 ex, 2 tbl

FIELD: biotechnology.

SUBSTANCE: claimed microarray represents ensemble of gel microcells on substrate made of glass, polymer, ceramic or composite material. Microcells contain immobilized procariotic or eucariotic cells. Microcells with immobilized cells are prepared using gel-forming solution including glycerol. Cellular microarray is used in living cell investigation. In this purpose cellular microarray is incubated in presence of marker, signal (e.g. cell fluorescence) is detected and according to signal level cell living function in microarray is evaluated.

EFFECT: simplified living cell investigation method without losses of cell viability.

12 cl, 5 dwg, 3 ex

FIELD: biology, medicine.

SUBSTANCE: invention relates to media used for reprogramming human and animal stem cells. Medium for the biochemical reprogramming human and animal stem cells containing the medium DI-MEM, fetal calve serum, insulin, 5-azacytidine, L-glutamine comprises additionally the medium F-12, dimethylsulfoxide, dexamethasone, hydrocortisone, N6-2'-dibutyryl-3',5'-cyclic adenosine monophosphate and ethanolamide in the required ratio of components. Invention provides enhancing the effectiveness in reprogramming stem cells in direction of cardiomyogenesis.

EFFECT: improved and valuable properties of medium.

2 dwg, 3 ex

FIELD: agricultural biotechnology, in particular, in vitro grape multiplication processes.

SUBSTANCE: method involves providing micro cutting of testing plants and planting thereof in liquid nutritive medium with reduced amount of macroelements and vitamins, with indole-acetic acid being added in an amount of 0.1-0.3 mg/l and emistim preparation with concentration of 10-7-10-10% being added into composition.

EFFECT: increased efficiency in multiplication of perspective sorts of grape sanitated from virus infection.

6 tbl

FIELD: biotechnology, medicine.

SUBSTANCE: peritoneal macrophages are placed in medium 1999 and subjected for effect of helium plasma obtained at current strength 30 A, voltage 20 V and gas consumption 2 l/min/ Cells are irradiated from distance 20 cm to plasmatron nozzle for 30 s. Method provides reducing time of physical factor effect on cells and allows carrying out the local effect on macrophage functions both in cultured cells and within the whole body. Invention can be used in clinical practice for stimulation of biological processes in cells and tissues.

EFFECT: improved method for stimulation.

1 tbl, 2 ex

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