Rapid diagnosis method for diagnosing human thyroid gland hyperfunction

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

 

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

The thyroid gland (thyroid) - an endocrine organ. Produced by it of thyroid hormones - thyroxine (T4) and triiodothyronine (T3) have high biological activity, regulate all kinds of exchanges, the synthesis of DNA, RNA, cell membrane permeability.

When overactive thyroid gland develops hyperthyroidism, characterized by persistent pathological increase in production of thyroid hormones. Excess hormones causes a disturbance in the functioning of most organs and body systems.

Among the methods of laboratory diagnosis of thyrotoxicosis emphasis on hormonal study. This identifies the levels of T3, T4 and thyroid stimulating hormone (TSH).

Key hormonal markers in the diagnosis of thyrotoxicosis are free thyroxine (SWT) and TSH. Concentration SWT is held in equilibrium by the mechanisms of regulation of the hypothalamic - pituitary - thyroid system and buffer capabilities linking tyrosinaemia globulin, albumin and prealbumin. Therefore, the concentration SWT not depends on the content tyrosinaemia globulin. Change SVT is an objective criteria each is evaluating the function of the thyroid gland.

Known immunodeficiency way to determine thyroid hormones Method based on the use of labeled antibodies. Due to its high specificity and sensitivity, provide monoclonal antibodies, can in-vitro to detect the presence of biological fluids of many substances (hormones, enzymes, opuholei markers etc). This category of methods includes radioimmunologically (RIA), enzyme-linked immunosorbent (ELISA) and immunofluorescence named because of the components and nature of the label. (Wood S.Y., Sokolovsky, Theory and practice radioimmunoassay. Guidance for staff laboratory services. “Globus”, Vienna, 1981; Reznikov A.G. Methods for the determination of hormones “Naukova Dumka”, Kiev, 1990).

Similar radioimmunoassay method, the fundamental principle of which is the competitive interaction of molecules of the hormone and specific antibodies or other associated proteins.

In the basis of the method is the use as a label radioactive isotope (wood S.Y., Sokolovsky, Theory and practice radioimmunoassay. Guidance for staff laboratory services. “Globus”, Vienna, 1981).

The main stages of the method:

- sample preparation

- the use of radioactive isotope

- the use of a gamma counter

The disadvantages of the method:

- the use of a radioactive isotope is

- the use of a gamma counter

- the cost of equipment

Prototype - immunoassay method for determination of free thyroxine in serum, plasma of human blood (application instruction set of reagents for qualitative assay of free thyroxine in serum, plasma. Registration Committee the public health Ministry. R/C No. 2000/93 from 06.04.2000. Set{FT4 EIA COBAS CORE. CT4 ELISA COBAS CORE}” Cat. No. 2051915/20736015, S508062).

The essence of the immunoassay method lies in the fact that the protein hormone ligand mark is not radioactive isotope, enzyme, in particular peroxidase from horseradish.

Peroxidase activity and, therefore, the content of conjugated to her hormone is measured by a photometric method using hydrogen peroxide as a substrate.

Implementation method:

The SEQUENCE of STATEMENT ANALYSIS MANUAL METHOD CT4 ELISA COBAS CORE

Bring all reagents to room temperature before use shake). To prepare the test tubes in duplicate (+1 for background samples).

1 incubation

To make reagents: 20 ál calibrator (Cs) or

20 ál control sera (4) or 20 μl of patient sample and

250 ál analytical buffer (5)

Add 1 bead

Thoroughly

Incubate for 30 minutes at 37°and With constant shaking

Rinse the washer Roche is about a given program

II incubation

To make 200 ál cold conjugate (2)

Incubate for 15 minutes in the refrigerator at 4-8°without shaking

Rinse in the washer for a given program

To prepare working substrate solution for 10 minutes before use rate of 1 volume of substrate solution TMB (8) + 4 amount of substrate buffer H2O2(10)

Add 250 μl of working substrate solution. Thoroughly

Incubate for 15 minutes at 37°and With constant shaking

Add 2 ml of sulfuric acid (12). Mix well.

To measure the absorption at 450 nm against FC, to calculate the concentration of ST4 using the ELA program on your computer.

The disadvantages of the method:

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

2. Immunodeficiency analysis is a complex study that require high skills and a thorough implementation of all procedures. Any small technical mistake due to the extremely high sensitivity of the tests and very low concentrations of detected substances leads to a distortion of the final result.

3. The high cost.

The objectives of the proposed invention are:

1. Raising awareness

2. Providing an Express release of thyroid hormones.

. The ability of the visual evaluation of an overactive thyroid gland.

For the development of the proposed invention, we used the possibility of registration of bio-energy radiation of living objects (in this case the thyroid gland).

It is known that a living object in all directions radiate the modulated electromagnetic radiation. Appear complex information encoded multifrequency patterns of oscillations, individual information and energy codes (Electromagnetic field in the biosphere. T1, 2. - M - 1984; Gulyaev YU.V., Year EE Physical fields of biological objects. In the book: Cybernetics of living. Biology and information - M - 1984 - P.39-60; Ananin SCI Bioenergy man - M - 1993).

Functioning cell is the source and carrier of complicated radiation, the cell structure may vary depending on the nature and frequency of the rhythm of radiation caused by the course of metabolic processes in the cell itself (Vpiznaju, Lpereira. Bioinformation function of natural electromagnetic fields. Novosibirsk, 1985).

The structure is a snapshot of the internal energy of interactions in a biological system (A.A. Malinovsky. Theory of structures and its place in the system approach. - M.: 1970 - C. - 10-31).

The technical novelty of the proposal is the physical examination of the thyroid gland of man, otlichayushiesya, which the glass plate is applied biological indicator is a mixture of amino acids, neurotransmitters and sulphate of magnesia, keeping it on the surface of the neck in the area of projection of the thyroid gland for 3-5 minutes, drying at T=+35-40°With 2-3 minutes, study in polarized light and in the presence of quartz compensator crystals, columnar, sestavino-strakhovatya, nepovinnyh and discharged polygon cameras motley painting diagnosed with a hyperactive thyroid gland.

To register energy radiation of living objects use a biological liquid crystals, which have a high optical activity (brown,, Walken J. Liquid crystals and biological structures - M - 1982). It is also used as a liquid crystal mixture of amino acids, neurotransmitters and sulphate of magnesia.

The method is as follows:

1. Prepare a biological indicator for the registration of the radiation of the thyroid gland, which is a model protokletos Association (MPKA), which use a mixture consisting of 0.1% aqueous solution of 10 essential amino acids (leucine, glycine, Proline, serine, phenylalanine, histidine, hydroxyproline, arginine, glutamic and aspartic amino acids), 0.5% aqueous solution of neuromediators - dopamine and histamine, 12% water dissolve the sulfate and magnesium). The ratio of amino acids, neurotransmitters and magnesium sulfate, respectively, 4:1:5.

2. Calibrated pipette biological indicator (BI) is applied to a glass plate in a thin film fluid volume 0,05-0,06 ml

3. Glass plate coated with BI is placed on the study area of the skin or areas of projection of the thyroid gland (lobe, isthmus), stand on the surface of the projection zones of the body for 3-5 minutes.

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

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

6. In the presence of drug crystals, columnar, sestavino-strakhovatya and discharged polygon cameras motley painting diagnosed with a hyperactive thyroid gland.

Here is the structure B represented by the model protokletos Association, photo 1. Visible subparallel aggregates made with grains and harness.

Photo 2 shows the structure resulting from the premises of the glass plate with BI and its standing for 3 minutes on the surface series of tablets synthesized L-thyroxine. Visible completed polygon camera white color.

Photo 3 shows the typical structure of BI obtained when placing a glass plate coated with BI, the area of projection of the thyroid of a healthy person. Look who s well oriented polygonal camera white color.

Examples

Example 1, photo 4,a, b. We investigated thyroid b-th And. DZ: Diffuse toxic goiter, the average degree of gravity, first identified. History (IB) No. 395. In photo 4,and the discharged polygonal camera motley painting, photo 4,b - snapvine crystals.

Technology. The glass plate caused 0.05 ml BI and rastegari” it in the form of a thin film. The plate was placed on the surface of the neck in the area of the projection of the thyroid gland and has stood for 5 minutes Then dried in a thermostat at T=+35°C for 2 min and examined under polarized light with a quartz compensator (QC). Visible discharged polygonal camera motley painting and snapvine crystals. At the same time in the UK b-th determined the content of the Holy T4, it was increased to 21 pmol/l (norm 16,1±0.18 pmol/l). The diagnosis of hyperthyroidism was confirmed.

Example 2, photo 5,a, B. Researched thyroid b-th A. DZ: Mixed toxic goiter, first identified, IB No. 289. In photo 5,and the discharged polygonal camera motley painting, photo 5,b - sestavino-Strahovanie crystals.

Technology. The glass plate caused 0.06 ml BI and placed the plate on the area of projection of the thyroid gland, has sustained its 3 minutes and Then dried in a thermostat at T=+38°C for 2 min and microscopically in polarized light with QC. Visible discharged polygonal camera motley painting is sestavino-Strahovanie crystals. In the study SVT in Sivaratri blood (SC) showed increased to 29 pmol/L. hyperfunction of the thyroid gland was confirmed.

Example 3, photo 6,a, b. Researched thyroid b-th 3. DZ: Diffuse toxic goiter, first identified. IB No. 218. In photo 6,and the discharged polygonal camera motley painting and snapvine crystals, photo 6,B.

Technology. The glass plate caused 0.06 ml BI and placed it on the neck in the area of the projection of the thyroid gland, has stood for 4 minutes Then dried in a thermostat at T=+38°C for 2 minutes, the Drug has microscopically in polarized light with QC. Are discharged polygonal camera motley painting and snapvine crystals. In SC b-th 3 has found an increased content SWT - 34 pmol/L. hyperfunction of the thyroid gland was confirmed.

Example 4, photo 7,a-century Explored the thyroid gland b-th W. DZ: Mixed toxic goiter. IB No. 24, photo 7,and the discharged polygonal camera motley painting, photo 7,b - columnar crystals.

Technology. The glass plate caused 0.06 ml BI, placed it on the area of projection of the thyroid gland, has stood for 5 min, dried in a thermostat at T=+40°C for 3 min and examined under polarized light with QC. Visible discharged polygonal camera motley painting and columnar crystals. At the same time in the UK determined SWT, he was raised - 33 pmol/L. Hyperfunk the thyroid gland was confirmed.

Example 5, photo 8A, b. researched thyroid b-th N. DZ: Mixed toxic goiter. IB No. 243. in photo 8,and the discharged polygonal camera motley painting, photo 8,b - snapvine crystals.

Technology. The glass plate caused 0.05 ml BI and placed it on the area of projection of the thyroid gland, endured 3 minutes and Then dried in a thermostat at T=+40°C for 3 min and examined under polarized light with QC. Visible discharged camera polygonal shape with the bright colouring and snapvine crystals. At the same time in the UK determined SWT, he was promoted and made 34 pmol/L. the Diagnosis of hyperthyroidism was confirmed.

Example 6, photo 9,a-century Explored the thyroid gland b-Oh D. DZ: Diffuse toxic goiter. IB No. 118. In photo 9,and the discharged polygonal camera white color. In photo 9,b - sestavino-Strahovanie crystals. In photo 9,in - snapvine crystals.

Technology. The glass plate caused 0.06 ml BI, placed it on the area of projection of the thyroid gland, endured 3 minutes and Then dried in a thermostat at T=+39°C for 3 minutes and microscopically in polarized light with QC. Discovered discharged camera polygonal shape with the bright colouring, sestavino-Strahovanie crystals and snapvine crystals. At the same time the SC has determined the level of St. T4, it was equal to 33 pmol/L. hyperfunction of the thyroid gland was confirmed.

Example 7, photo 10A-century Experiments is whether the thyroid gland b-th O. DZ: nodular toxic goiter. IB No. 284.

Photo 10A shows a depleted polygonal camera motley painting, photo 10B, in - columnar crystals.

Technology. The glass plate caused 0.05 ml BI, stood on the surface of the neck, in the area of the projection of the thyroid gland for 4 min, dried in a thermostat at T=+39°C for 3 min and examined under polarized light with QC. Found empty polygon camera motley painting and columnar crystals. At the same time the SC has determined the level of St. T4, he was promoted and was 40 pmol/L. hyperfunction of the thyroid gland was confirmed.

Example 8, photo 11,a, b. researched thyroid b-nd HP HD: Mixed toxic goiter. IB No. 11. Photo 11 and discharged polygonal camera motley painting and sestavino-Strahovanie crystals, photo 11,B.

Technology. The glass plate caused 0.06 ml BI, stood 5 min in the zone of projection of the thyroid gland was dried in a thermostat at T=+40°C for 3 min and examined under polarized light with QC. Visible discharged polygonal camera motley painting and sestavino-Strahovanie crystals. The IC has determined the level of St. T4, he was raised to 30 pmol/L. hyperfunction of the thyroid gland was confirmed.

Example 9, photo 12,a, b, C. Investigated thyroid b-nd, IB No. 939. DZ: Mixed toxic goiter. In photo 12,and visible discharged polygonal camera motley painting, photo 12,b - Shestov is about-Strahovanie crystals.

Technology. The glass plate caused 0.06 ml BI, placed on the area of projection of the thyroid gland, stood 5 min, dried in a thermostat at T=+37°C for 3 min and examined under polarized light with QC. Are discharged polygonal camera motley painting and sestavino-Strahovanie crystals. At the same time the SC has determined the level of St. T4, he was promoted and was equal to 46 pmol/L. hyperfunction of the thyroid gland was confirmed.

Example 10, photo 13,a, b. Researched thyroid b-th M DZ: Diffuse toxic goiter. IB No. 239. Photo 13,as shown empty polygon camera motley painting, photo 13,b - columnar crystals.

Technology. The glass plate caused 0.05 ml BI, placed in the area of the projection of the thyroid gland, has withstood 4 min, dried in a thermostat at T=+35°C for 2 min and examined under polarized light with QC. Visible discharged polygonal camera motley painting and columnar crystals. At the same time in the UK determined SWT, he was raised - 33 pmol/l

Hyperfunction of the thyroid gland was confirmed.

Just spent 515 research.

The advantages of the proposed method:

1. Using a highly sensitive liquid biological indicator, responsive to the release of excessive amounts of thyroxine, a hormone of the thyroid gland.

2. The native characteristics of bacterial and physiology of the conducted research the Oia.

3. High information content.

4. The reduction of material costs, the simplification of the basic method. Medical-biological effect of the application of the method is to carry out rapid diagnosis of an overactive thyroid gland that can be used for mass investigations.

The method of rapid diagnosis of hyperthyroidism person, including physical examination of the thyroid gland, characterized in that the glass plate is applied biological indicator is a mixture consisting of 0.1% aqueous solution of amino acids: leucine, glycine, Proline, serine, phenylalanine, histidine, hydroxyproline, arginine, glutamic and aspartic, 0.5% aqueous solution of neurotransmitters: dopamine, and histamine, 12% aqueous solution of sulfuric acid magnesium in the ratio of amino acids as neurotransmitters : sulfuric acid magnesium = 4:1:5, the led is kept on the surface of the neck in the area of the projection thyroid gecesi for 3-5 min, dried at T=+35-40°With, examined in polarized light with a quartz compensator and in the presence of crystals, columnar, sestavino-strakhovatya, nepovinnyh and discharged polygon cameras motley painting diagnosed with a hyperactive thyroid gland.



 

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