RU2305840C1 - Method for instant diagnosis of fatty hepatosis - Google Patents

Abstract

FIELD: medicine, physiology.

SUBSTANCE: claimed method includes preparation of biological indicator comprising of 0.1 % aqueous solution of amino acid mixture, such as asparaginic acid, glutamic acid, glycine, thryptophan, valine, leucine, proline arginine, threonine, serine; 0.5 % dophamine aqueous solution; 0.25 % DNA sodium salt aqueous solution; 12 % magnesium sulfate aqueous solution in ratio of 3:1:1:5. Indicator is applied on in volume of 0.01-0.02 ml in form of path on 6-7 glass plates. Plates are placed on skin surface of right hypochondrium in liver projection region, hold for 2-3 min, dried at T=+18-20°C and liver irradiation is investigated in polarized light with quartz compensator. In case of presence in preparation deformed polygonal chambers, and/or roundish, and/or ovaloid, and/or rhomboid ones with or without processes, inserts of multicolor staining fatty hepatosis is diagnosted.

EFFECT: non-injury improved method for instant diagnosis of liver disorders associated with fatty hepatosis.

Description

The present invention relates to medicine, gastroenterology, pathophysiology, biology, and can be used to diagnose fatty hepatosis (GH).

ZhG - (fatty degeneration of the liver; fatty infiltration; steatosis of the liver) is a chronic disease characterized by an increase in the accumulation of fat in hepatocytes (A.I. Strukov, V.V. Serov. Pathological anatomy. M., 1979).

GH can occur as an independent disease or as a syndrome against other diseases, for example, alcoholism, diabetes mellitus, Cushing's syndrome, myxedema, malabsorption syndrome (V.T. Ivashkin, Yu.O. Shulpekova. Non-alcoholic steatohepatitis // Digestive diseases. - 2000. - No. 2. - p. 41-45).

The main morphological and functional unit of the liver is the hepatic lobule formed by hepatocytes (Fig. 1).

Lipocytes - fat-storing liver cells - are located in the perisinusoidal space, have numerous long processes that penetrate deep between hepatocytes, (figure 2 - functional morphology of the liver. Lipocyte with a large number of lipid inclusions in the perisinusoidal space, x8500).

Lipocytes contain mitochondria with a light matrix and a small amount of cristae (Serov V., Lapin K. Morphological diagnosis of liver diseases. M., 1989. S.29-30).

In hypertension, small drops of fat initially accumulate in the hepatocyte cytoplasm, merging subsequently into larger ones or into one fatty vacuole, which occupies the entire cytoplasm and shifts the nucleus to the periphery of the cell. Fat vacuoles of several cells can merge with each other and form fatty cysts (A. S. Loginov, L. I. Aruin. Clinical morphology of the liver. M., 1985. S. 34). Figure 3 shows the preparation of fatty liver disease - pronounced fatty degeneration of pericentral hepatocytes. Coloring Sudan III-IV. x200; large-drop fatty degeneration, fatty cysts around the central vein. Van Gieson stain. x504.

The liver is the target organ in violation of lipid metabolism, combining into a lipid distress syndrome (V.S. Saveliev, E.G. Oblokov, V.A. Petukhov. Lipid distress syndrome in surgery // Bull. Experiment. Biology and medicine . - 1999. t.127. - No. 6. - p. 604-611). The accumulation of fat in liver cells may result from an increase in the intake of free fatty acids (FFA), a decrease in the rate of β-oxidation of FFA in the mitochondria, excessive synthesis or secretion of very low density lipoproteins (VLDL).

An important role in disrupting the metabolism of FFA belongs to alcohol. An additional risk factor for GI is an excess of fat in the diet combined with a protein deficiency. Factors involved in the formation of hypertension: drugs (glucocorticoids, amiodarone, estrogens, tetracycline, non-steroidal anti-inflammatory drugs), extended resection of the small intestine, gastroplasty for obesity, rapid weight loss, prolonged parenteral nutrition, excess bacterial colonization of the small intestine, α-alpha -lipoproteinemia, limb lipodystrophy, Wilson-Konovalov disease, Weber-Christian disease. ZhG can accompany chronic hepatitis C, especially caused by a virus of the 3rd genotype (A.O. Bueverov, M.V. Mayevskaya. Some pathogenetic and clinical issues of non-alcoholic steatohepatitis // Clin. Prospects for gastroenterology, hepatology. - 2003. - No. 3. - p. 2-7).

The possibilities for identifying a VH are different. These include a puncture biopsy of the liver with the study of histopreparations (V. Serov, K. Lapshin. Morphological diagnosis of liver diseases. M., 1989. S.29-30), instrumental studies of the liver - ultrasound diagnostics (V.V. Mitkov. Clinical guidance by UZD. M., 1996. S.35-38, S.52-58), tomography - (R.I. Gabunia, E.K. Kolesnikova. Computed tomography in clinical diagnostics. M., 1995. S.352 )

The analogue. Morphological study of liver biopsy samples (Loginov A.S., Aruin L.I. Clinical morphology of the liver. M., 1985. S. 5-16). Biopsy methods: percutaneous puncture, transvenous, sighting (during laparoscopy) and marginal (during surgery). The biopsy is obtained by puncture needles or special tweezers, then it is processed:

- fixed in 10% formalin buffered by the Lily method;

- pour the material into paraffin, from each block prepare up to 25-30 slices, mounting them on 4-5 slides;

- sections stained with hematoxylin and eosin, as well as picrofuxin according to the Van Gieson method.

The last stage is the microscopy of the drug.

The disadvantages of the method.

- trauma to the patient (invasiveness of the study);

- the need for a large number of technical means;

- the use of needles, laparoscopes, ultrasound machines;

- the use of a series of chemicals for the preparation of material;

- the duration of execution, which does not allow for operational monitoring of treatment.

Ultrasound examination of the liver using a sensor with a frequency of 3.5-5 MHz is accepted as the closest analogue (V.V.Mitkov. Clinical Guide to Ultrasound Diagnostics. M., 1996, v. 1, S.35-38, 52-58) .

The essence of the method is based on the study of the echostructure of the liver using an ultrasound scanner.

The study is as follows:

1. The patient is placed on his back.

2. The sensor is installed parallel to the right costal arch and the angle of inclination of the sensor is gradually changed.

3. The state of the liver is assessed by determining its contour, shape, size, echogenicity, structure, vascular pattern.

The disadvantages of the method:

- the use of an ultrasound scanner that affects the physiological state of the subject and increases the cost of the study;

- mutual influence on the image of the liver of surrounding organs, which reduces the reliability of the results of the study;

- occurrence of artifacts during scanning;

- stimulation of the growth of some neoplasms in the liver (cyst, hemangioma, etc.).

The objectives of the invention:

1. Ensuring differentiation of the diagnosis of fatty hepatosis depending on the severity of the pathological process.

2. Increasing the information content of diagnostics by visualizing the results of diagnostics.

3. Operational monitoring of the effectiveness of treatment.

4. The exclusion of invasiveness and radiation in the diagnosis of fatty hepatosis.

The technical result of the proposal is the rapid registration of liver energy radiation in the form of certain images of a biological liquid crystal, which allows you to quickly register dynamics in the liver caused by fatty hepatosis, differentiate these disorders, completely eliminate the morbidity and reduce the cost of diagnosis.

A significant novelty of the proposal is the registration of liver radiation using a biological indicator (BI), for which, unlike the known method (ultrasound of the liver), a biological indicator (a mixture of a 0.1% aqueous solution of amino acids - asparagine, glutamine, glycine, tryptophan, valine, leucine, proline, arginine, threonine, series; 0.5% dopamine neurotransmitter aqueous solution; 0.25% DNA sodium salt solution; 12% 3: 1: 1: 5 aqueous solution of magnesium sulfate, which is applied to 6-7 glass plates, place them on and the skin surface of the right hypochondrium in the zone of the projection of the liver is kept for 2-3 minutes, dried at T = + 18-20 ° C, examined in polarized light with a quartz compensator and in the presence of deformed polygonal chambers and / or round ones, and / or ovaloid, and / or rhomboid, with processes and without processes, inclusions of motley color diagnose fatty hepatosis.

The method is as follows.

1. Prepare a biological indicator (BI) - energy-informational biological test system (model) for recording liver radiation. A mixture is prepared consisting of a 0.1% aqueous solution of 10 amino acids (aspartic, glutamine, glycine, tryptophan, valine, leucine, proline, arginine, threonine, serine; 0.5% aqueous solution of the neurotransmitter dopamine; 0.25% sodium salt solution DNA; 12% aqueous solution of magnesium sulfate in a ratio of 3: 1: 1: 5, respectively.

2. A BI pipette with a volume of 0.01-0.02 ml is applied onto a glass slide in the form of a track.

3. Slides (glass plates) with BI applied on them are placed on the skin surface of the right hypochondrium, maintained for 2-3 minutes, dried at T = + 18-20 ° C for 2-3 minutes.

4. The preparations are examined in polarized light with a quartz compensator, and if the preparations have deformed polygonal chambers and / or round, and / or ovaloid, and / or rhomboid, with processes and without processes, inclusions of motley color, fatty hepatosis is diagnosed.

We present data on BI (energy-information model of radiation of a biological object) - a variant of a biological liquid crystal that responds to radiation by the appearance of a specific structure.

Information radiation from various objects (living, biocosal, inert) is one of the general principles of energy-informational interaction in nature.

Information influence is understood as a specific reaction of systems to a weak material-energy stimulus, perceived as a signal in the form of a code (N.F. Raimes. A systematic approach to solving the problem of the interaction of a medium with a biological object. In the book: Electromagnetic fields in the biosphere, vol. 1 ., C.294-300, M., 1984).

Structure - a snapshot of the internal interactions in the system (A.A. Malinovsky. Theory of structure and its place in the systems approach. In the book: System Studies. Yearbook. M., 1970, pp. 10-31).

We have developed a BI (a variant of a biological liquid crystal, an energy-information matrix). The proposed model is a complex biological system with self-organization and high spatial orientation. The sodium salt of DNA, dopamine, amino acids that form the cell nucleus (DNA and RNA), which are part of it, are themselves sources of information, have the ability to perceive information about the electron-conformational interactions occurring in other substances, and transmit information in the form of structures. Liquid crystals are used as biosensor devices. Liquid crystalline DNA and amino acids - analogues of cell associations - “read” the information-energy code of energetic biological radiation, which results in the appearance of a certain structure (Yu.M. Evdokimov. Liquid-crystalline forms of nucleic acids // Bulletin of the Russian Academy of Sciences. - 2003. - № 8. - S.712-721; S.D. Varfolomeev, Yu.M. Evdokimov, M.A. Ostrovsky. Sensory biology, sensory technologies and the creation of new human senses // Bulletin of the Russian Academy of Sciences. - 2000. - No. 2. p. 99-108).

We used BI to register the biological radiation of the liver based on the data that modulated bioelectromagnetic radiation is emitted from a person (his organs) in all directions. Complex information-coded multi-frequency oscillation patterns arise, wave radiation forming individual information-energy codes (V.L. Vvedensky, V.I. Ozhogin. Supersensitive magnetometry and biomagnetism. M., 1986; Sh.M. Kogan, Physical nature of the biofield man. In the book: Anomalous phenomena, facts, studies, hypotheses. M., 1991., issue 1. S.24-37).

Figure 4 shows the initial structure of the BI, the cellular type of assembly of the drug is visible.

In FIG. Figure 5 shows the structures of BI obtained by placing glass plates with BI applied on them on the liver zone of a healthy person. The plates were held for 3 minutes, dried in a thermostat at Т = + 18 ° С for 3 minutes, and examined in polarized light with a quartz compensator (CC). There are oriented polygonal chambers, the appearance of which resembles hepatocytes (Fig. 1).

The proposed method is compared with known methods for determining fatty hepatosis. A total of 318 studies were conducted. It was revealed that the statistical significance was p <0.05.

Example 1, Fig.6. B. K. Diagnosis: Fatty hepatosis. Diabetes mellitus, type II. Case history No. 398. Figure 6 shows the structure of the BI obtained by placing glass plates (SP) on the skin of the right hypochondrium in the area of the projection of the liver.

There are deformed polygonal chambers with rounded and rhomboid inclusions without processes of variegated color.

Technology. 0.02 ml of BI were applied to 6 glass plates (SP) and placed in the right hypochondrium in the area of the projection of the liver, kept for 2 minutes, then dried in a thermostat for 3 minutes at T = + 20 ° C and examined in polarized light with CC.

There are deformed polygonal chambers with rounded and rhomboid inclusions of variegated color. At the same time, a liver biopsy was performed with a biopsy and liver ultrasound examination.

Biopsy: pronounced zonal fatty degeneration, medium-droplet fat inclusions predominate, mainly centrolobular, droplet diameter up to 10-12 microns.

Ultrasound: Liver PD - 152 mm (N <140 mm). LD - 74 mm (N <65 mm).

The form is ordinary.

The contours are smooth.

Echogenicity is common.

The structure is homogeneous, fine-grained.

The bile ducts are not dilated.

Portal vein - 10.5 mm

The vascular pattern is normal.

In 6, 7 segments of the liver along the visceral surface, zones of increased echogenicity are determined - 1.5 × 2 cm, 2.3 × 3.5 cm, of a homogeneous fine-grained structure, with uneven contours.

Conclusion: ultrasound - signs of focal changes in the liver. Fatty hepatosis.

The diagnosis of hypertension was confirmed.

Example 2, Fig.7. B. N. Diagnosis: fatty hepatosis of alcoholic etiology. Case history No. 18. Figure 7 shows the structure of the BI, there are deformed polygonal chambers, inclusions are rounded, rounded, rhomboid, with processes and without variegated color.

Technology: 0.01 ml of BI in the form of a track were applied to the joint venture and placed in the right hypochondrium, in the area of the projection of the liver, kept for 3 minutes, then dried in an thermostat at T = + 19 ° C for 2 minutes, examined in polarized light with QC. Deformed polygonal chambers were found, inclusions - oval, round, rhomboid, with processes and without processes, of mottled color.

A biopsy and an ultrasound of the liver were performed simultaneously.

Biopsy: severe steatosis, hepatocytes are filled with fat, which is located in the central and intermediate sections of the lobules (3rd - 2nd acinus zones). Individual hepatocytes contain large fat vacuoles.

Ultrasound scan:

Liver PD 164 mm (N <140 mm). LD 92 mm (N <65 mm).

The form is ordinary.

The contours are uneven.

Echogenicity - increased.

The structure is fine-grained.

The bile ducts are not dilated.

Portal vein - 12 mm.

The vascular pattern is depleted.

Conclusion: ultrasound - signs of diffuse changes in the liver (fatty hepatosis).

The diagnosis of hypertension was confirmed.

Example 3, Fig. 8. B-naya M. Diagnosis: Fatty hepatosis of drug origin (exposure to tetracycline). Medical history No. 279.

Fig. 8 shows the structure of the BI — the deformed polygonal chambers with rounded, diamond-shaped inclusions of variegated color, with processes and without processes are visible.

Technology: 0.02 ml of BI in the form of a track were applied to the joint venture, placed in the right hypochondrium, in the zone of the projection of the liver, kept for 3 minutes, dried in an thermostat at Т = + 20 ° С for 2 minutes, examined in polarized light with CC . Deformed polygonal chambers with rounded, rhomboid inclusions of variegated color, with and without processes, were found.

A biopsy and an ultrasound of the liver were done simultaneously.

Biopsy: Hepatocytes with signs of fatty degeneration were found, fatty vacuoles are located in more than 1/3 of the liver cells (second degree of fatty degeneration).

Ultrasound scan:

Liver PD 146 mm (N <140 mm). LD - 57 mm (N <65 mm)

Form - plain

Contours - Smooth

Echogenicity is common.

Structure - homogeneous, fine-grained

The bile ducts are not dilated.

Portal vein - 10 mm

The vascular pattern is normal.

At the border of 7, 8 liver segments along the diaphragmatic surface, a zone of increased echogenicity is determined - 1.7 × 2.3 cm, with uneven contours, of a fine-grained structure.

Conclusion: ultrasound - signs of focal changes in the liver. Fatty hepatosis.

The diagnosis of hypertension was confirmed.

Example 4, Fig.9. B-th J. Diagnosis: fatty hepatosis of alcoholic origin. Case history No. 345. In Fig.9 there are polygonal deformed chambers with rounded inclusions of variegated color, with processes.

Technology: 0.01 ml of BI in the form of a track were applied to the joint venture, placed in the right hypochondrium, in the zone of the projection of the liver, kept for 2 minutes, dried in an thermostat at Т = + 18 ° С for 2 minutes, studied in polarized light with CC . Found deformed polygonal chambers with rounded inclusions of variegated color, with processes. A liver biopsy and an ultrasound scan were done at once.

Biopsy: Alcoholic steatosis is expressed, hepatocytes are filled with fat, merge, form cysts. Lipids are found in macrophages of portal tracts.

Ultrasound scan:

Liver PD - 148 mm (N <140 mm). LD - 61 mm (N <65 mm).

The form is ordinary.

The contours are smooth.

Echogenicity is common.

The structure is fine-grained

The bile ducts are not dilated.

Portal vein - 9 mm.

The vascular pattern is normal.

In 6, 7 segments of the liver, a zone of increased echogenicity of 4.5 × 6 cm, with an uneven contour of a homogeneous, fine-grained structure, is determined by the visceral surface.

Conclusion: ultrasound signs of focal changes in the liver. Fatty hepatosis.

The diagnosis of hypertension was confirmed.

Patient J. was prescribed a course of treatment with ursodeoxycholic acid (UDCA) at a dose of 350 mg / day for 5 months. UDCA is a hydrophilic bile acid with anticholestatic, cytoprotective and immunomodulating properties.

After a course of treatment, a control rapid diagnosis of structural changes in the liver was performed using the BI developed by us. On figa shows the structure of the BI obtained by registering the radiation of the liver of the patient W .: there are cameras of a polygonal shape (normal version).

At the same time, an ultrasound of the liver was performed.

Liver: normal form.

Echogenicity - not increased.

The bile duct is not dilated.

Portal vein - 14 mm.

The left lobe is 65 mm.

Right lobe 140 mm.

The contours are even, the structure is homogeneous, the vascular pattern is ordinary.

Conclusion: no pathology was detected.

No liver biopsy data revealed any pathology.

Example 5, Fig. 10. B-th A. Diagnosis: fatty hepatosis. Obesity III degree. Case history No. 16. In figure 10 there are deformed polygonal chambers with rounded, diamond-shaped and ovaloid inclusions of variegated color, with processes and without.

Technology: 0.02 ml of BI in the form of a track were carried to the joint venture, placed in the right hypochondrium, in the zone of the projection of the liver, kept for 3 minutes, dried in an thermostat at T = + 19 ° C for 3 minutes, studied in polarized light with CC . Found deformed polygonal chambers with rounded, rhomboid inclusions of motley color, with processes and without processes.

Immediately performed a biopsy and ultrasound of the liver.

Biopsy: large-drop fatty degeneration of the II degree. The diameter of fat droplets is up to 21 microns, fat cysts are found.

Ultrasound: liver PD - 187 mm (N <140 mm). LD - 92 mm (N <65 mm).

The form is ordinary.

The contours are uneven.

Echogenicity - increased.

The structure is fine-grained.

The bile ducts are not dilated.

Portal vein - 10 mm.

The vascular pattern is depleted.

Conclusion: Uz - signs of diffuse changes in the liver (fatty hepatosis).

The diagnosis of hypertension was confirmed.

Example 6 of FIG. 11. B-naya I. Diagnosis: Fatty hepatosis due to the administration of uncorrectable doses of prednisolone. Case history No. 181. In Fig. 11, there are deformed polygonal chambers, with rounded, oval-shaped inclusions of variegated color, with processes and without processes.

Technology: 0.01 ml of BI in the form of a track were applied to the joint venture, placed in the right hypochondrium, in the zone of the projection of the liver, kept for 2 minutes, dried in an thermostat at Т = + 20 ° С for 2 minutes, studied in polarized light with CC .

Found deformed polygonal chambers with rounded and oval-shaped inclusions of variegated color, with processes and without. Immediately performed a biopsy and ultrasound of the liver.

Biopsy: Macrovesicular steatosis detected. The cell cytoplasm is filled with one large drop, which shifts the cell nucleus to the side.

Ultrasound: liver PD - 146 mm (N <140 mm). LD - 76 mm (M <65 mm).

The form is ordinary.

The contours are smooth.

Echogenicity is common.

The structure is homogeneous, fine-grained.

The bile ducts are not dilated.

The portal vein is 10.5 mm.

Vascular pattern is reinforced.

In 4, 7, 8 segments of the liver, a zone of increased echogenicity, an irregular shape, with a total size of 5.5 × 7 cm, with uneven contours, and a homogeneous structure is determined.

Conclusion: Uz - signs of local changes in the liver. Fatty hepatosis.

The diagnosis of fatty hepatosis was confirmed.

Example 7, Fig. 12. B. D. Diagnosis: Fatty hepatosis. Severe hypothyroidism. Case history No. 38.

Figure 12 shows the deformed polygonal chambers with rhomboid inclusions of motley color without processes.

Technology: 0.02 ml of BI in the form of a track were applied to the joint venture, placed in the right hypochondrium, in the zone of the projection of the liver, kept for 3 minutes, dried in an thermostat at Т = + 18 ° С for 3 minutes, studied in polarized light with CC . Found deformed polygonal chambers with rhomboid inclusions of variegated color without processes.

A liver biopsy and an ultrasound scan were done at once.

Biopsy: More than 1/3 of hepatocytes contain large fatty vacuoles, individual diameter up to 25 microns. Fatty steatosis of the II degree.

Ultrasound scan

Liver PD - 148 mm (N <140 mm). LD - 65 mm (N <65 mm).

The form is ordinary.

The contours are smooth.

Echogenicity is common.

The structure is homogeneous, fine-grained.

The bile ducts are not dilated.

Portal vein - 11 mm.

The vascular pattern is normal.

In 5, 6, 7 segments of the liver, a zone of irregular shape, increased echogenicity, a total size of 6 × 4.5 cm, with uneven contours of a fine-grained structure is determined.

Conclusion: Uz - signs of local changes in the liver. Fatty hepatosis.

The diagnosis of hypertension was confirmed.

The advantages of the proposed method.

The use of a highly sensitive biological indicator - a marker of impaired liver structure in case of fatty hepatosis - allows to provide high information content and efficiency, nativeness and physiology, non-invasiveness of the method.

Compared with the basic methods for diagnosing fatty hepatosis, the study time is reduced by 30-40%, and material costs - by 65%.

Biomedical and social effect of the method lies in its accessibility and the possibility of conducting screening studies.

Claims (1)

A method for the rapid diagnosis of fatty hepatosis, characterized in that a biological indicator is preliminarily prepared consisting of a 0.1% aqueous solution of a mixture of amino acids - aspartic, glutamic, glycine, tryptophan, valine, leucine, proline, arginine, threonine, serine; 0.5% dopamine aqueous solution; 0.25% aqueous solution of sodium salt of DNA, 12% aqueous solution of magnesium sulfate in a ratio of 3: 1: 1: 5, which is applied in a volume of 0.01-0.02 ml in the form of a track on 6-7 glass plates, place them on the skin surface of the right hypochondrium in the zone of the projection of the liver, incubated for 2-3 minutes, dried at T = + 18-20 ° C, in the polarized light with a quartz compensator, the radiation of the liver is examined, and if the preparation has deformed polygonal chambers, and / or rounded, and / or oval, and / or rhomboid, with processes and without processes, inclusions of motley aski, diagnose fatty hepatosis.

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