RU2247375C1 - Rapid diagnosis method for diagnosing human thyroid gland hyperfunction - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves applying biological indicator as mixture composed of 0.1% aqueous solution of amino acids: leucine, glycine, proline, serine, phenylalanine, histidine, oxyproline, arginine, glutamic amino acid and aspartic amino acid, 0.5% aqueous solution of neuromediators like dopamine and histamine, 12% aqueous solution of magnesium sulfate in proportion of amino acids : neuromediators : magnesium sulfate = 4:1:5. Indicator kept on neck surface in thyroid gland projection during 3-5min. It is dried at T=+35-40°C, studied in polarized light with quartz compensator. Columnar, columnar-and-striated, sheaf-like crystals and discharged polygonal variegated chambers being observed, thyroid gland hyperfunction is to be diagnosed.

EFFECT: high accuracy of diagnosis.

13 dwg

Description

The alleged invention relates to medicine, endocrinology, pathophysiology, biology and can be used to diagnose hyperfunction (thyrotoxicosis) of the thyroid gland.

The thyroid gland (thyroid gland) is an endocrine organ. The thyroid hormones produced by it - thyroxine (T4) and triiodothyronine (T3) possess high biological activity, regulate all types of metabolism, DNA synthesis, RNA, and permeability of cell membranes.

With an increase in thyroid function, thyrotoxicosis develops, characterized by a persistent pathological increase in the production of thyroid hormones. Excess hormones cause a malfunction in most organs and systems of the body.

Among the methods of laboratory diagnosis of thyrotoxicosis, hormonal research is of the greatest importance. In this case, the levels of T3, T4 and thyroid stimulating hormone (TSH) are determined.

The key hormonal markers in the diagnosis of thyrotoxicosis are free thyroxine (St. T4) and TSH. The concentration of St. T4 is kept in equilibrium by the regulation mechanisms of the hypothalamic - pituitary - thyroid system and the buffer capacity for binding of thyroxin-binding globulin, albumin and prealbumin. Therefore, the concentration of St. T4 does not depend on the content of thyroxin-binding globulin. Change St. T4 is an objective criterion for evaluating thyroid function.

A known immunometric method for determining thyroid hormones is based on the use of labeled antibodies. Due to its high specificity and sensitivity provided by monoclonal antibodies, it is possible to detect in vitro the presence of many substances in biological fluids (hormones, enzymes, tumor markers, etc.). This category of methods includes radioimmunological (RIA), enzyme immunoassay (ELISA) and immunofluorescence, which received their names due to the components and nature of the label. (Wood UG, Sokolovsky G. Theory and practice of radioimmunoassay. A guide for laboratory personnel. Globus, Vienna, 1981; Reznikov AG Methods for the determination of hormones Naukova Dumka, Kiev, 1990).

An analogue is a radioimmunological method, the fundamental principle of which is the competitive interaction of hormone molecules and a specific antibody or other bound protein.

The method is based on the use of a radioactive isotope as a label (Wood UG, Sokolovsky G. Theory and practice of radioimmunoassay. Manual for laboratory personnel. Globus, Vienna, 1981).

The main steps of the method:

- sample preparation

- use of a radioactive isotope

- use of gamma counter

The disadvantages of the method:

- use of a radioactive isotope

- use of gamma counter

- high cost of equipment

The prototype is an enzyme-linked immunosorbent assay for determining free thyroxine in serum, human blood plasma (Instructions for using a reagent kit for high-quality immunoassay analysis of free thyroxin in serum, human plasma. Registration Board of the Ministry of Health of the Russian Federation. R / U No. 2000/93 from 04/06/2000. Set “{FT4 EIA COBAS CORE. CT4 ELISA COBAS CORE}” Cat. No. 2051915/20736015, S508062).

The essence of the enzyme immunoassay is that the protein hormone ligand is not labeled with a radioactive isotope, but with an enzyme, in particular horseradish peroxidase.

The activity of peroxidase and, therefore, the content of the hormone conjugated with it is measured photometrically using hydrogen peroxide as a substrate.

The implementation of the method:

MANUAL STATEMENT OF ANALYSIS BY MANUAL METHOD CT4 ELISA COBAS CORE

Bring all reagents to room temperature (shake well before use). Prepare duplicate tubes (+1 for background sample).

1 incubation

Dispense reagents: 20 µl of recalibrator (3c) or

20 μl of control serum (4) or 20 μl of a patient sample and

250 μl assay buffer (5)

Add 1 ball

To stir thoroughly

Incubate for 30 minutes at 37 ° C with constant shaking.

Rinse with Roche washer according to the set program

II incubation

Dispense 200 μl Cold Conjugate (2)

Incubate for 15 minutes in a refrigerator at 4-8 ° C without shaking.

Rinse with washer according to the set program

Prepare a working solution of the substrate 10 minutes before use, based on the calculation: 1 volume of the substrate solution of TMB (8) + 4 volumes of the substrate buffer H ₂ O ₂ (10)

Add 250 μl of the working solution of the substrate. To stir thoroughly

Incubate for 15 minutes at 37 ° C with constant shaking.

Add 2 ml of sulfuric acid (12). To stir thoroughly.

Measure the absorbance at 450 nm against AF, calculate the concentration of CT4 using the ELA program on a computer.

The disadvantages of the method:

1. The specificity of the samples is not absolute. It is possible that cross-reacting substances can falsify results.

2. Immunometric analysis is a complex study that requires highly qualified personnel and the thorough implementation of all procedures. Any minor technical oversight due to the extremely high sensitivity of the tests and very low concentrations of the substances being determined leads to a distortion of the final result.

3. The high cost.

Objectives of the alleged invention:

1. Increasing information

2. Providing express registration of the release of thyroid hormones.

3. The ability to visually assess thyroid hyperfunction.

To develop the alleged invention, we used the ability to register bio-energy radiation from living objects (in this case, the thyroid gland).

It is known that modulated electromagnetic radiation is emitted from a living object in all directions. Complex information-encoded multi-frequency oscillation patterns, individual information-energy codes (Electromagnetic field in the biosphere. T1, 2. - M. - 1984; Gulyaev Yu.V., Godik E.E. Physical fields of biological objects. In the book: Cybernetics) biology and information - M. - 1984 - S.39-60; Ananin V.F. Bioenergetics of man - M. - 1993).

A functioning cell is the source and carrier of complex radiation, the cell structure can vary depending on the nature and frequency of the radiation rhythm due to the course of metabolic processes in the cell itself (V.P.Kaznacheev, L.P. Mikhailova. Bioinformation function of natural electromagnetic fields. Novosibirsk, 1985).

Structure - a snapshot of the internal energy interactions in the biological system (A.A. Malinovsky. Theory of structures and its place in the systematic approach. - M .: 1970 - S. - 10-31).

The technical novelty of the proposal is a physical study of the human thyroid gland, characterized in that a biological indicator is applied to the glass plate - a mixture of amino acids, neurotransmitters and magnesium sulfate, keeping it on the surface of the neck in the projection area of the thyroid gland for 3-5 minutes, drying at T = + 35-40 ° C for 2-3 minutes, the study in polarized light and in the presence of crystals with a quartz compensator - columnar, columnar-streaky, sheaf-like and discharged polygonal mottled color chambers diagnose thyroid hyperfunction.

To register the energy radiation of living objects, biological liquid crystals are used that have high optical activity (Brown G., Walken J. Liquid crystals and biological structures - M. - 1982). We also used a mixture of amino acids, neurotransmitters, and magnesium sulfate as a liquid crystal.

The method is as follows:

1. Prepare a biological indicator for detecting thyroid radiation, which is a model of protocellular association (MPCA), which uses a mixture of 0.1% aqueous solution of 10 vital amino acids (leucine, glycine, proline, serine, phenylalanine, histidine, oxyproline , arginine, glutamic and aspartic amino acids), 0.5% aqueous solution of neurotransmitters - dopamine and histamine, 12% aqueous solution of magnesium sulfate). The ratio of amino acids - neurotransmitters and sulfate magnesia, respectively, 4: 1: 5.

2. A biological indicator (BI) is applied by a pipette to a glass plate in the form of a thin film, the liquid volume is 0.05-0.06 ml.

3. A glass plate with a BI applied on it is placed on the studied area of the skin or the projection zone of the thyroid gland (lobes, isthmus), kept on the surface of the projection zone of the organ for 3-5 minutes.

4. The drug is dried in a thermostat at T = + 35-40 ° C for 2-3 minutes.

5. The sample is examined in polarized light with a quartz compensator.

6. If there are crystals in the preparation — columnar, columnar-streaky, and discharged polygonal chambers of variegated color — hyperthyroidism is diagnosed.

We present the structure of the BI represented by the protocellular association model, photo 1. Subparallel aggregates made with grains and bundles are visible.

Photo 2 shows the structure obtained by placing a glass plate with BI and keeping it for 3 minutes on the surface of a row consisting of synthesized L-thyroxine tablets. One can see the completed polygonal cameras of variegated coloring.

Figure 3 shows the typical structure of the BI obtained by placing a glass plate with the applied BI on the thyroid projection zone of a healthy person. One can see well-oriented polygonal cameras of variegated color.

We give examples

Example 1, photo 4, a, b. The thyroid gland was studied. I. DZ: Diffuse toxic goiter, moderate severity, first detected. Medical history (IB) No. 395. Photo 4, a shows discharged polygonal cameras of variegated color, and photo 4, b shows sheaf-like crystals.

Technology. 0.05 ml BI was applied to the glass plate and it “spread” in the form of a thin film. The plate was placed on the surface of the neck in the area of the thyroid projection and held for 5 minutes. Then it was dried in a thermostat at Т = + 35 ° С for 2 min and examined in polarized light with a quartz compensator (CC). Visible discharged polygonal chambers of motley color and sheaf-like crystals. At the same time, the content of sv. T4, it turned out to be elevated - 21 pmol / L (normal 16.1 ± 0.18 pmol / L). The diagnosis of thyrotoxicosis was confirmed.

Example 2, photo 5, a, b. The thyroid gland was examined in A. DZ: Mixed toxic goiter, first detected, IB No. 289. Photo 5, a shows discharged polygonal cameras of variegated color, and photo 5, b shows columnar-streaky crystals.

Technology. 0.06 ml of BI was applied to the glass plate and the plate was placed on the thyroid projection zone, it was held for 3 minutes. Then it was dried in a thermostat at Т = + 38 ° С for 2 min and microscopied in polarized light with CC. Visible discharged polygonal chambers of motley color and columnar-hatching crystals. In the study of St. T4 in the blood serum (SC), an increase of 29 pmol / L was found. Hyperfunction of the thyroid gland was confirmed.

Example 3, photo 6, a, b. Thyroid b-th was examined. 3. DZ: Diffuse toxic goiter, first detected. IS No. 218. Photo 6, a shows discharged polygonal cameras of motley color and sheaf-like crystals, photo 6, b.

Technology. 0.06 ml of BI was applied to the glass plate and placed on the neck in the area of the thyroid projection, kept for 4 minutes. Then it was dried in a thermostat at Т = + 38 ° С for 2 minutes. The drug was microscopied in polarized light with QA. There are rarefied polygonal chambers of motley coloring and sheaf-like crystals. In SC b-th 3, an increased content of St. T4 was found - 34 pmol / L. Hyperfunction of the thyroid gland was confirmed.

Example 4, photo 7, a-c. Investigated thyroid b-th S. DZ: Mixed toxic goiter. IS No. 24, in photo 7, a shows discharged polygonal cameras of variegated color, in photo 7, b, c - columnar crystals.

Technology. 0.06 ml of BI was applied to the glass plate, placed on the thyroid projection zone, kept for 5 minutes, dried in a thermostat at Т = + 40 ° С for 3 minutes, and studied in polarized light with CC. Visible discharged polygonal chambers of motley color and columnar crystals. At the same time, the level of St. T4 was determined in the UK; it turned out to be increased - 33 pmol / L. Hyperfunction of the thyroid gland was confirmed.

Example 5, photo 8a, b. investigated thyroid b-th N. DZ: Mixed toxic goiter. IS No. 243. on photo 8, a discharged polygonal cameras of variegated color are shown, on photo 8, b - sheaf crystals.

Technology. 0.05 ml of BI was applied to the glass plate and placed on the thyroid projection area, held for 3 minutes. Then it was dried in a thermostat at Т = + 40 ° С for 3 min and studied in polarized light with CC. Visible discharged polygonal chambers of motley color and sheaf-like crystals. At the same time, the level of St. T4 was determined in the UK; it turned out to be increased and amounted to 34 pmol / L. The diagnosis of thyrotoxicosis was confirmed.

Example 6, photo 9, a-c. They investigated the thyroid gland of the second D. DZ: Diffuse toxic goiter. IS No. 118. Photo 9, a shows discharged polygonal cameras of variegated color. In photo 9, b - columnar-streaky crystals. On a photo 9, in - sheaf-shaped crystals.

Technology. 0.06 ml of BI was applied to the glass plate, placed on the thyroid projection zone, held for 3 minutes. Then it was dried in a thermostat at Т = + 39 ° С for 3 minutes. and microscopied in polarized light with KK. Discharged polygonal chambers of motley color, columnar-hardened crystals and sheaf-like crystals were found. At the same time in the UK determined the level of St. T4, it was 33 pmol / L. Hyperfunction of the thyroid gland was confirmed.

Example 7, photo 10a-c. Investigated thyroid b-th O. DZ: toxic toxic goiter. IS No. 284.

Photo 10a shows discharged polygonal chambers of variegated color, photo 10b, c shows columnar crystals.

Technology. 0.05 ml BI was applied to the glass plate, kept on the surface of the neck, in the area of the thyroid projection for 4 minutes, dried in a thermostat at T = + 39 ° C for 3 minutes, and studied in polarized light with CC. Discharged colored polygonal chambers of motley coloring and columnar crystals were found. At the same time in the UK determined the level of St. T4, it was increased and amounted to 40 pmol / L. Hyperfunction of the thyroid gland was confirmed.

Example 8, photo 11, a, b. investigated thyroid b-th L. DZ: Mixed toxic goiter. IS No. 11. Photo 11, a shows discharged polygonal cameras of variegated color and columnar-hatching crystals, photo 11, b.

Technology. 0.06 ml BI was applied to the glass plate, held for 5 min in the area of the thyroid projection, dried in a thermostat at Т = + 40 ° С for 3 min, and studied in polarized light with CC. Visible discharged polygonal chambers of motley color and columnar-hatching crystals. In SK determined the level of St. T4, it was increased - 30 pmol / L. Hyperfunction of the thyroid gland was confirmed.

Example 9, photo 12, a, b, c. Investigated thyroid g. B. G. IB No. 939. DZ: Mixed toxic goiter. In photo 12, a discharged polygonal cameras of variegated color are visible, in photo 12, b, c - columnar-hatching crystals.

Technology. 0.06 ml of BI was applied to the glass plate, placed on the thyroid projection zone, held for 5 minutes, dried in a thermostat at Т = + 37 ° С for 3 minutes, and studied in polarized light with CC. There are discharged polygonal chambers of variegated color and columnar-hatching crystals. At the same time in the UK determined the level of St. T4, it was increased and was equal to 46 pmol / L. Hyperfunction of the thyroid gland was confirmed.

Example 10, photo 13, a, b. They investigated the thyroid gland of the second M. DZ: Diffuse toxic goiter. IS No. 239. Photo 13, a shows discharged polygonal cameras of variegated color, photo 13, b shows columnar crystals.

Technology. 0.05 ml BI was applied to the glass plate, placed in the thyroid projection zone, held for 4 minutes, dried in an oven at T = + 35 ° C for 2 minutes, and studied in polarized light with CC. Visible discharged polygonal chambers of motley color and columnar crystals. At the same time, the level of St. T4 was determined in the UK; it turned out to be increased - 33 pmol / L.

Hyperfunction of the thyroid gland was confirmed.

A total of 515 studies were conducted.

The advantages of the proposed method:

1. The use of a highly sensitive liquid crystal biological indicator that responds to the release of an excess of thyroxine - a thyroid hormone.

2. The nativeness and physiology of the study.

3. High information content.

4. Reduction of material costs, simplification of the basic method. The biomedical effect of the application of the method consists in the rapid diagnosis of thyroid hyperfunction, which can be used in mass studies.

Claims (1)

A method for the rapid diagnosis of hyperthyroidism of a person, including physical examination of the thyroid gland, characterized in that a biological indicator is applied to the glass plate - a mixture consisting of 0.1% aqueous solution of amino acids: leucine, glycine, proline, serine, phenylalanine, histidine, hydroxyproline, arginine, glutamine and aspartic, 0.5% aqueous solution of neurotransmitters: dopamine and histamine, 12% aqueous solution of sulfuric acid magnesia in the ratio: amino acids: neurotransmitters: sulfuric acid magnesia = 4: 1: 5 , the indicator is kept on the surface of the neck in the projection area of the thyroid glands for 3-5 minutes, dried at T = + 35-40 ° C, examined in polarized light with a quartz compensator and in the presence of crystals - columnar, columnar-streaky, sheaf-like and discharged polygonal chambers of mottled color diagnose hyperthyroidism.

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