The method of determining the degree of damage to the cells of the body

 

(57) Abstract:

The method of determining the degree of damage of the cells of the body can be used in medicine and veterinary medicine. The degree of damage to the cells is determined in the presence of the drug "Clairin", treating the leukocytes of peripheral blood gamma radiation. Measure the frequency of sister chromosome exchanges. Determine the protection factor3the value radioadaptive answer RAO and the degree of damage to the cells Cpaccording to the formula Cp=K3/RAO. Diagnosis of the degree of damage of the cells in the presence of the drug "Clairin" at the present methodology allows to obtain more complete information about the state of the organism. 2 C.p. f-crystals, 1 table.

The invention relates to the field of biochemistry, and in particular to methods of diagnostics of cellular structures.

Known methods of diagnostics of cells in presence of various substances in the extracellular fluid, the number of echinocytes in the blood, etc. /1-3/. Their drawback is the narrow scope limited to specific types of cells that does not allow a General assessment of the condition of the whole body or specific types of mutagenic effects.

Cdetermined by the formula

< / BR>
where Pmand Pto- frequency SAS after treatment with mutagen and without treatment (control), respectively, and Pziand PCaccordingly, the frequency fell after sequential processing of a protective agent (interferon) and mutagenic, as well as a protective agent only.

The disadvantage of this method is incomplete information about the state of the organism.

Problem to be solved in the framework of the proposed method was a modification of the prototype method so that you can judge ispolzovaniem as a protective agent of the drug "Clairin", use as a mutagenic factor of ionizing radiation mode, the calling radioadaptive response of cells RAO, determination of the values of RW and KCin the presence of Blarina", and the degree of cell damage Cpaccording to the formula Cp= KC/RAO, diagnosing that the body is healthy or has a low degree of damage when Cp= 1.0 to 2.0, the average degree of damage when Cp>2.0 and severe damage when the negative value of the UES and/or the absence of cell growth in the presence of the drug "Clairin".

The research was carried out at the introduction of Blarina" in a nutrient medium at a dose of not less than 110-7g/l for at least 12 hours before exposure.

To obtain RAO irradiation carried out first in small doses of ionizing radiation /0,1-10 RAD/, and then large doses of 2-4 G/ according to the method described in /5/.

This RAO is determined by the formula

< / BR>
where Pwith- the amount of damage the sequential irradiation of small and large doses of gamma rays, Pb- the amount of damage after irradiation of cells with large dose of gamma-rays; Pto- the amount of damage control without irradiation. RAO - radioadaptive response, reflecting the decrease in strelecky small doses of radiation.

The basis of the invention lies stated by the authors of property drug "Clairin" block in the system of blood lymphocytes besttransport cells phytohemaglutinin if the damage of the genome. At the same time it was found that the drug with reversible disorders of the membranes and/or cell walls are able to return the cells lost their properties, almost to the level of the indicators of healthy cells.

Currently, the drug "Clairin" get extraction callus tissue Ungernii Victor.

The value of Cpwhen the diagnosis was obtained by aggregation of indicators representative groups of patients with different pathologies.

The analysis was carried out as follows. As a source of small doses of radiation used beta radiation H3-thymidine. (0.1 IWC/ml) for 20 hours. As a large doses were used gamma-irradiation at a dose of 2 Gy.

The blood lymphocytes were cultured in vitro according to standard methods. Stimulation (besttransport) lymphocytes was performed by means of phytohemagglutinin (PHA). Cells were irradiated with gamma rays at a dose of 2 Gy in 20 hours after the beginning of cultivation and recorded after 52 hours. Analyzed the frequency and spectrum of chromosome aberrations (S) before irradiation. This experimental system allows to differentiate the effects of intracellular exposure of incorporated radionuclides external gamma irradiation. OH chromating type are the result of exposure to radionuclides in the S phase of the mitotic cycle; OH chromosome type is the result of external exposure radiointerview radiation in the G-phase, i.e., gamma irradiation in 20 hours after stimulation of cells with PHA.

The experiments showed the possibility of using the drug in doses of not lower than 110-7g/l when processing time is not less than 12 hours, but the effects of the drug has the limit values a big error.

The most effective mode is the dose 110-6g/l for 16 hours. The use of large quantities of the drug over a longer processing time is not economically feasible.

Industrial applicability of the method is illustrated by the following examples.

Example 1. The study was performed on the blood cells of children as clinically healthy donors (6), and patients (10 people) living within 3-9 years in territories with contamination levels of from 3.2 to 26.3 CI/km in the accident at the Chernobyl nuclear power plant (Bre methodology with different quantities of the drug. The results are given in table 1.

Literature

1. A. C. the USSR 1028716, CL C 12 N 15/00.

2. Gg Of Avtandil. Medical morphometry in the diagnosis of disease, M, 1983, S. 6-120.

3. Auth.St. The USSR 924102, CL C 12 N 1/00.

4. Jakubowski E. L. et al. Genetics, XXIV, No. 11, 1988, S. 2050.

5. Olivieri G. et al., Science, 1984, v. 228, p. 594-597.

1. The method of determining the degree of damage to the cells of the body, involving the cultivation of peripheral blood leukocytes tested in the presence of a protective agent and without it, the treatment of cells with gamma radiation, the measurement of the frequency of sister chromosome exchanges and determination of the coefficient of protection, characterized in that as a protective agent drug use "Clairin", processing of leukocytes carry doses of ionizing radiation, causing radioadaptive response cells, while additionally determine the amount radioadaptive response of cells of the test and determine the degree of cell damage WITHpaccording to the formula Cp= KC/RW, where KC- the coefficient of the protection when using the drug "Clairin", RAO - radioadaptive cell response, WITHpthe degree of cell damage, diagnosing that the body practice is at Cpmore than 2.0, and the presence of severe damage negative values radioadaptive answer or absence of cell growth in the presence of the drug "Clairin".

2. The method according to p. 1, characterized in that Blairin" is introduced into the medium at a concentration of 1 to 10-5- 1 10-7mg/ml, at least 12 h before irradiation.

3. The method according to PP.1 and 2, characterized in that the irradiation is carried out sequentially with radiation doses of 0.1 - 10 rad and 2 - 4 Gr.

 

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