RU2296326C2 - Method for predicting acute intoxication - Google Patents

Abstract

FIELD: medicine, laboratory diagnostics.

SUBSTANCE: the present innovation deals with preparing erythrocytic hemolyzate due to diluting washed out erythrocytes in distilled water at the ratio of 1:1 to be kept for about 1-2 h at 20-25°C followed by applying it onto a slide's surface at the quantity of 0.01-0.03 ml and dried for about 24-27 h at 18-30°C and relative environmental moisture of 55-60%. One should carry out microscopic survey and at detecting fissures in the space of central and peripheral areas of dried drop of amorphous substances it is possible to diagnose acute intoxication. Application of the present method enables to establish the presence of the marker of acute toxemia of different etiology in earlier terms and match therapy that provides acceleration of further recovery.

EFFECT: higher accuracy and efficiency of diagnostics.

6 dwg, 1 ex

Description

The invention relates to medicine, in particular to laboratory research methods, and can be used to diagnose acute toxemia of various etiologies.

A known method for the diagnosis of endogenous intoxication, which according to the morphological picture of blood serum allows you to determine the marker of endogenous intoxication, presented in the form of "toxic plaques" placed on the field of a dried drop of blood serum. (Shabalin V.N., Shatokhina S.N. Morphology of human biological fluids. - M., Publishing House. Chrysoston, 2001, - p. 93, 118, 122).

In this method, the method of wedge-shaped dehydration was used, which ensures the conversion of blood serum to the solid phase, which allows early determination of the systemic organization of blood serum and the identification of a marker specific for chronic intoxication. However, it is not possible to identify acute endogenous intoxication by this method.

The clinical manifestations of endogenous toxemia, especially in the acute period, are well known and are characterized by a pronounced violation of the vital functions of the body. However, a marker of acute toxemia of various etiologies has not yet been identified.

It has been established that under extreme and pathological conditions the nature of the interaction of various components of biological fluids at the molecular level changes significantly. The endotoxins entering the blood are able to enter into easily reversible bonds with molecules embedded in the membranes, changing their permeability, structure, and, in particular, it is known that erythrocyte membranes bind blood plasma endotoxins.

In accordance with this, an erythrocyte hemolysate (EG) consisting of an intracellular erythrocyte fluid and their lysed membranes was chosen as a biological fluid for the study.

A known method for the diagnosis of acute endogenous intoxication, including the preparation of a hemolysate of red blood cells by diluting washed red blood cells in distilled water and its study (US Pat. RF No. 2217753, CL G 01 N 33/49, 2002).

The disadvantages of this method are:

- limited functionality of the method, due to the fact that with its help determine endogenous intoxication only with burns;

- the need to use expensive technical means, complex chemical reagents and mathematical calculations to determine endogenous intoxication with burns.

In addition, this method involves the introduction of an animal into anesthesia before causing injury (burn), therefore, can only be used in experiment.

In accordance with this, the task was set to expand the functionality of the method by identifying in the early stages a marker of acute intoxication of various etiologies, which, together with the necessary complex of intensive care, will additionally include the necessary detoxification complex, which is often crucial for patients with acute toxemia of various etiologies ( shock condition, coma due to acute drug intoxication).

To solve this problem, in a method for the diagnosis of acute endogenous intoxication, including the preparation of erythrocyte hemolysate by diluting washed red blood cells in distilled water and its study, it is proposed that the dilution be carried out in a ratio of 1: 1, kept for 1-2 hours at a temperature of 20-25 ° C, then apply on the surface of the slide in the amount of 0.01-0.03 ml, dry for 24-27 hours at a temperature of 18-30 ° C and a relative humidity of 55-60%, conduct microscopic examination and When exposed to cracks in the central and peripheral zones of a dried drop of amorphous substances, diagnose acute intoxication.

Figure 1 shows the facies of the erythrocyte hemolysate of the patient without the manifestation of shock.

Figure 2 is an enlarged fragment of figure 1

Figure 3 shows the erythrocyte hemolysate facies of a patient with traumatic shock.

In Fig.4 is an enlarged fragment of Fig.3 with an arrow indicating an amorphous substance in the crack space of a dried drop.

Figure 5 is an enlarged fragment of the facies of a narcological patient with an arrow indicating an amorphous substance in the space of a crack of a dripped drop.

Figure 6 is an enlarged fragment of the facies of a narcological patient with an arrow indicating an amorphous substance in the crack space of a dried drop.

The method is as follows.

Washed red blood cells obtained from unstabilized venous and arterial blood of the peripheral sections of the vascular bed are subjected to a given osmotic dilution with distilled water in a ratio of 1: 1, the resulting solution is kept for 1-2 hours at a temperature of 20-25 ° C, which is necessary for the destruction of red blood cell membranes . Ready standard erythrocyte hemolysate (absolutely homogeneous, bright ruby red color, not forming any suspended particles and inclusions with vigorous shaking) is investigated by the method of wedge-shaped dehydration. In this case, an erythrocyte hemolysate in an amount of 0.01-0.03 ml, in the form of a drop, is applied to the surface of a glass slide, dried for 24-27 hours at a temperature of 18-30 ° C and a relative humidity of 55-60%, then carry out a microscopic examination using a stereo microscope MZ-12 "Leica" (Germany) and if a dried drop of amorphous substances is detected in the space of cracks in the central and peripheral zones, acute intoxication is diagnosed.

A specific example of the implementation of the method.

Patient O. 53 years old, East. disease X-8295, was admitted to the intensive care unit with a diagnosis of closed fractures of 4, 5, 6. 7, 8 ribs on the left. Closed fracture of the right clavicle. Open fracture of the left humerus, bones of the left leg. Closed fragmentation fracture of the bones of the right tibia in the lower third with an offset. Traumatic shock.

In blood tests noted: stress glycosuria, increased ESR and creatine. The level of hemoglobin, red blood cells, lymphocytes and the total protein content are determined at the level of the lower limit of normal. A study of the proposed method. Microscopy revealed: the erythrocyte hemolysate facies is represented by an unstable protein-salt type of systemic organization and is characterized by the presence of wide radial, arcade or three-beam cracks. In the inner space of almost all cracks in the central zone, an amorphous substance is defined. X-ray microanalysis of this substance was carried out. It is established that it has an organic origin. In the intensive care unit, active anti-shock therapy and therapeutic measures aimed at normalizing homeostasis have been started. As a result of the therapeutic measures, the general condition of the patient stabilized. Microscopic evaluation of the erythrocyte hemolysate facies noted that the type of systemic organization becomes stable and the amount of amorphous substance in the cracks is significantly reduced. This circumstance is objective evidence of a decrease in endogenous intoxication.

We examined 129 patients hospitalized in a state of shock of various etiologies. Of these, 49 patients (37.98%) were hospitalized in a state of traumatic shock; 9 patients (6.97%) were hospitalized in a coma due to acute drug intoxication; 19 patients (14.72%) were hospitalized in a state of alcoholic coma (acute alcohol intoxication). In all the studied samples in patients of this category, regardless of the origin of the shock, an amorphous substance was found in the cracks of the central and peripheral zones of the dried drop (Figs. 3, 4, 5, 6).

When studying the morphological picture of the facies of the EG of patients with traumatic disease without manifestations of shock and healthy individuals, the absence of any inclusions in the cracks of the zones of the dried drop was found (Figs. 1, 2).

Therefore, the amorphous substance inside the cracks of the central and peripheral zones of the dried drop can serve as a marker of acute intoxication of various etiologies.

The organic nature of this substance was revealed by X-ray spectral microanalysis (RSMA). The morphology of the studied object was studied in backscattered electrons. The average content and ratio of chemical elements in the pathological formation inside the cracks, measured in mass percent C (49.2), N (2.76), O (13.04), Cl (0.82), Na (0.27 ), S (0.23), K (0.91), Fe (0.76), indicated that this substance is formed mainly by organic matter with inclusions of mineral components.

It was established that the pathological substance in the space of cracks in the facies of the EG persists for a long time, but as anti-shock detoxification therapy and therapeutic measures aimed at normalizing homeostasis are carried out, the number of cracks with pathological substance is significantly reduced.

Thus, when studying the morphological picture of intracellular fluid - erythrocyte hemolysate - in patients with acute toxemia of various etiologies, a marker of acute intoxication, represented by a pathological substance of organic origin, localized in the cracks of the facies, was first discovered.

The use of this method will allow in the early stages to establish the presence of a marker of acute toxemia of various etiologies and determine the treatment that accelerates recovery.

Claims (1)

A method for diagnosing acute endogenous intoxication, including the preparation of erythrocyte hemolysate by diluting washed red blood cells in distilled water and its study, characterized in that the dilution is carried out in a ratio of 1: 1, kept for 1-2 hours at a temperature of 20-25 ° C, after which applied to the surface of a glass slide in an amount of 0.01-0.03 ml, dried for 24-27 hours at a temperature of 18-30 ° C and relative humidity of the environment 55-60%, microscopic examination is carried out and if t Eshina central and peripheral zones of the dried droplet amorphous substances diagnose acute intoxication.

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