RU2578029C2 - Diagnostic technique for steatohepatitis - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention represents a diagnostic technique for non-alcoholic steatohepatitis, involving analysing patient's saliva, differing by the fact that a patient's saliva drop is added with a drop of the crystal-forming substance - 3% alcohol copper chloride; the produced preparation is kept for 24 hours in a horizontal position at temperature 20-25°C, humidity 50-70%, away from direct sunlight and heating appliances and examined under the microscope; presence and intensity of non-alcoholic steatohepatitis is detected by a presence and amount of amorphous dark-yellow mass overlaying the broom- or beam-shaped crystals in the microscopic preparation, whereas absence of non-alcoholic steatohepatitis is indicated by the broom- or beam-shaped crystals with no amorphous dark-yellow mass overlaid.

EFFECT: implementing the invention enables the fast non-invasive diagnosis of steatohepatitis.

2 tbl, 2 dwg

Description

This method relates to medicine, and in particular to the diagnosis of steatohepatitis.

Steatohepatitis - inflammatory changes against the background of fatty liver. Fatty liver degeneration has a number of equivalent synonyms: liver steatosis, steatohepatosis, fatty hepatosis, fatty liver.

Criteria for the diagnosis of steatohepatitis.

Microscopic examination of a biopsy (liver tissue) obtained by percutaneous liver biopsy reveals signs of balloon dystrophy of hepatocytes, steatonecrosis and lobular inflammation (represented by intralobular infiltrates consisting of polymorphic leukocytes, lymphocytes and mononuclear phagocytes). In most cases, large-droplet steatosis is detected, mainly centrilobular, characterized by the presence of large single lipid droplets in the hepatocyte cytoplasm with a displacement of the nucleus to the periphery of the lobule (“Non-alcoholic fatty liver disease in questions and answers.” Mehtiev SN, Mekhtieva OA, Zinoviev E.N., St. Petersburg State Medical University named after Academician I.P. Pavlov, study guide, 2011, p. 26). However, a significant obstacle to the widespread use of this method is its cost, additional technical and professional costs.

In addition, a percutaneous liver biopsy has a number of significant drawbacks:

- This is an invasive method;

- inconvenience to the patient;

- there are contraindications for use;

- it is impossible for a short period of time, due to the threat of complications of surgical manipulation, to control;

Currently, there is no one laboratory research method that can reliably confirm or exclude steatohepatitis.

However, non-invasive methods are used in medicine for a comprehensive assessment of the degree of steatohepatitis activity: determination of hepatocyte apoptosis markers (cytokeratin-18) in blood serum, determination of the concentration of pro-inflammatory cytokines (interleukins-6, tumor necrotizing factor index, adiponectin, highly sensitive C-reactive protein), determination of markers oxidative stress (reactive substances of thiobarbituric acid, oxidation of low density lipoproteins, general level of lipid peroxides) ("Non-alcoholic rovaya liver disease: Questions and Answers. "Mehdiyev SN, Mehdiyev OA, Zinoviev EN State Medical University. Academician Pavlov, textbook, 2011, p. 29). Disadvantages: the determination of a single marker cannot indicate the presence of steatohepatitis, since a change in these indicators is possible with other diseases.

The purpose of this invention is to simplify the diagnosis of steatohepatitis. The closest analogue of this method is the "Method for the diagnosis of insulin resistance", invention No. 20112112724/14 (019167) from 08/28/13, the Method allows to detect metabolic disorders in the body at an early stage. A drop of a biological substrate (saliva) of the patient and a drop of a crystal-forming substance / 0.9% sodium chloride solution are applied to a glass slide.

When examining a drop under a microscope, the presence and severity of insulin resistance are determined by the presence and quantity in the crystallogram of roundish irregular crystals in the field of view against the background of clear diamond-shaped crystallization centers. In the absence of crystals in the entire field of view, the absence of insulin resistance is judged.

We have developed a method for the diagnosis of steatohepatitis, which consists in the following. A drop of biological substrate (saliva) and one drop of an alcoholic 3% solution of copper chloride (CuCl ₂ ) are applied to a glass slide. The drug is kept for 24 hours in a strictly horizontal position at room temperature (20-25 ° C), humidity 50-70%, away from direct sunlight and heating devices. Microscopy of the preparation is carried out using a Biolam light microscope at a magnification of 7X8.

Based on the distribution of crystalline elements, non-alcoholic steatohepatitis is diagnosed. The presence of steatohepatitis is characterized by the appearance in the microscopic preparation of crystals in the form of “panicles” or “bunches” with the application of amorphous dark yellow masses (3) (Fig. 1). The absence of non-alcoholic steatohepatitis is characterized by the presence in the microscopic preparation of crystals in the form of “panicles” (1) or “bunches” (2) without imposing amorphous dark yellow masses (Fig. 2).

The essential features characterizing the invention:

1) saliva crystallography is used to detect non-alcoholic steatohepatitis;

2) a crystal-forming substance is used - a 3% alcohol solution of copper chloride;

3) in the presence of non-alcoholic steatohepatitis in the microscopic preparation amorphous dark yellow masses appear, superimposed on the crystals in the form of “panicles” or “bunches”;

4) the number of amorphous masses can determine the severity of non-alcoholic steatohepatitis.

Signs 2, 3, 4 are distinctive from the closest analogues.

The main advantages characterizing the invention:

- simplicity of the method;

- cheapness, minimal material costs;

- speed of crystallographic research (after 24 hours the result is obtained);

- not laborious;

- non-invasive;

- does not cause inconvenience to the patient;

- the ability to perform repeated studies, to monitor and correct treatment.

Research results.

Studies were carried out in the departments of hepatology, psychiatry, gastroenterology of the Main Clinical Military Hospital of the FSB of Russia, and the crystalline structures of saliva were studied in patients with chronic non-alcoholic steatohepatitis, alcoholic liver disease and in healthy patients (Table 1). The clinical diagnosis is confirmed by laboratory, instrumental methods of research, histological examination of liver tissue.

In the described manner, a study was conducted in 80 patients, which was confirmed by the act of implementation No. 436/11 of 03.10.11, “Method for the diagnosis of steatohepatitis”, approved by the head of the SCWG of the FSB of the Russian Federation. As a result of the study, a correlation was first made between the revealed crystal structures: amorphous masses superimposed on the “broom”, and hyperfermentemia, a histological examination of liver tissue; for the first time, a relationship was found between the change in the amount of amorphous masses superimposed on the “broom” in the preparation and the indicators of blood hyperenzyme (Table 2).

This method is simple to implement and allows you to identify metabolic disorders at an early stage. Since October 2011, after completing the examination of 80 patients, this method continues to be used in the hepatological and endocrinological departments of the hospital. Over the past period, 183 patients were examined.

Thus, our proposed method for the diagnosis of steatohepatitis differs from other methods in that a drop of a crystal-forming substance (3% alcohol solution of copper chloride) is added to a drop of saliva of the patient, the resulting preparation is kept for 24 hours in a horizontal position at a temperature of 20-25 ° C, humidity 50-70%, away from direct sunlight and heating devices and examined under a microscope. The presence of non-alcoholic steatohepatitis is determined by the presence in the microscopic preparation of amorphous dark yellow masses (3) superimposed on crystals in the form of “panicles” (1) or “bunches” (2) (Fig. 1). The absence of non-alcoholic steatohepatitis is determined by the presence in the microscopic preparation of crystals in the form of “panicles” (1) or “bunches” (2) without imposing amorphous dark yellow masses (Fig. 2). The method allows with minimized material costs and ease of implementation to diagnose metabolic disorders of the body.

Claims (1)

A method for the diagnosis of non-alcoholic steatohepatitis syndrome, which allows to detect metabolic changes in the body, including a study of the patient’s saliva, characterized in that a drop of a crystal-forming substance, 3% alcohol solution of copper chloride, is added to a drop of saliva of the patient, the preparation is kept for 24 hours in a horizontal position at a temperature 20-25 ° C, humidity 50-70%, away from direct sunlight and heating devices and examined under a microscope, while the presence and severity of non-alcoholic eatohepatitis is determined by the presence and quantity of amorphous dark yellow masses in a microscopic preparation superimposed on crystals in the form of “panicles” or “bunches”, and the absence of non-alcoholic steatohepatitis is determined by the presence in the microscopic preparation of crystals in the form of “panicles” or “bunches” without amorphous dark yellow masses.

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