RU2424521C1 - Diagnostic technique for burn toxemia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: patient's blood serum in volume 100-200 mcl is put on a slide, and a preparation is dehydrogenated in a warm air flow at temperature 40-50°C and humidity 20-30% for 15-20 minutes in a horizontal position. A facies is matched with a burn toxemia pattern by the values: a structural property index, crystallisability, a type of crystal and amorphous structures interaction, a degree of facies desctruction, a uniformity of crystalline and amorphous facies elements distribution, a degree of facies porosity manifestation, manifestation of separate crystallisation zones, manifestation of a boundary protein band and structural relief. If observing at least six of nine criteria values matched with the pattern, burn toxemia is diagnosed.

EFFECT: increased diagnostic accuracy, reduced analysis time.

2 tbl, 1 ex, 2 dwg

Description

The invention relates to medicine, in particular to laboratory diagnostics, and can be used in combustiology for the diagnosis of burn toxemia.

In recent years, an important role has been given to the study of endogenous intoxication syndrome. It was shown that endotoxemia develops in all pathological conditions associated with increased catabolism or blockade of the body's detoxification systems. Protein toxins, medium-molecular peptides, are considered to be the substrate responsible for the occurrence of local pathological effects of endogenous intoxication in renal failure. The latter included endogenous components with a molecular weight of 500-2000 Da, which are formed during proteolysis in damaged tissues, as well as in the plasma itself when proteolytic enzymes enter the bloodstream. For many types of pathology, the functioning of the protease and antiprotease systems is characteristic, and as a result of the activation of proteolysis, a large number of protein degradation products with a molecular weight of 300-5000 Da are accumulated. In domestic literature, the class of medium molecular weight proteolysis products is referred to as medium weight molecules (MSM). The chemical composition of MSM is very heterogeneous and combines peptides, glycopeptides, nucleopeptides, endorphins, amino sugars, polyamines, polyhydric alcohols, some humoral regulators (insulin, glucagon, some vitamins, nucleotides, oligosaccharides, glucuronic acid derivatives and others).

The study of the pathogenesis and clinical picture of burn disease made it possible to show that endogenous intoxication also plays a leading role in their formation. Therefore, verification of its presence and severity is necessary to select the tactics for maintaining this patient population.

For the prototype of the invention, a well-known method for the diagnosis of burn toxemia is selected, including a study of blood serum (Malakhova M.Ya. Method for recording endogenous intoxication. A manual for doctors. St. Petersburg., 1995. - P.7-8).

The known method is as follows. Venous blood is collected from the patient and plasma is obtained from it. The sample is photometric at wavelengths from 238 to 300 nm. The obtained spectrograms are analyzed and the level of toxic products formed during normal and impaired metabolism is determined.

However, this method has the following disadvantages. The method allows to obtain only fragmentary information about the state of the body and therefore requires a series of additional studies. This circumstance lengthens the examination period of the patient and significantly reduces its diagnostic significance. In addition, it is time-consuming and requires expensive equipment to complete.

The objective of the invention is to increase the accuracy of diagnosis, providing the possibility of express diagnostics, reducing research time and simplifying the method.

The problem is solved in that in the known method for the diagnosis of burn toxemia, including the study of blood serum, the patient's blood serum in a volume of 100-200 μl is applied to a glass slide and the drug is dehydrated in a stream of warm air at a temperature of 40-50 ° C and a humidity of 20- 30% for 15-20 minutes in a horizontal position, the obtained crystalloscopic facies is compared with the burn toxemia pattern by indicators: structural index, crystallizability, type of interaction of crystalline and amorphous structures, the degree of degradation of the facies, the uniform distribution of crystalline and amorphous elements of the facies, the degree of severity of the facies cellularity, the severity of individual crystallization zones, the severity of the edge protein zone and the texture relief and, if at least six of the nine criteria coincide with the pattern, they diagnose burn toxemia.

The present invention meets the criteria of "novelty" and "inventive step", since the conducted patent information research did not reveal the sources of patent and scientific and technical literature that would discredit the novelty of the proposed method, as well as known methods with significant features of the proposed technical solution.

The authors of the invention carried out research work related to the assessment of the information content of biocrystallization. To solve these problems, a study was made of the features of free crystallization of blood serum in 35 patients who had burn toxemia, the presence of which was confirmed by classical methods (the level of molecules of average weight, total protein and its fractions, creatinine and blood urea, bilirubin, myoglobin), including " gold standard ”- by determining the level of medium-weight molecules in the blood. Assessment of the results of crystallogenesis of biosubstrates was carried out using the system of criteria described below. Based on these studies, a crystalloscopic pattern was formed (Table 1), for which a diagnostic value was approved.

Table 1 The pattern of free crystallization of serum of patients with endotoxemia Indicator Value Structural index 1-2 Crystallizability 2-3 Type of interaction of crystals and amorphous bodies 0 The degree of destruction of the facies 1-2 Uniform distribution of sample elements 2-3 The severity of cellularity 3-4 Facialization 0-1 The severity of the marginal zone 1-2 Sharpness texture sample 2-3

To perform a qualitative and quantitative assessment of the results of intrinsic crystallization of blood serum, the following criteria are used:

1. The structural index. Estimated by a 4-point system:

0 points - the presence in the micropreparation of only an amorphous substance;

1 point - the discrete part of the sample is represented by amorphous bodies and single crystals;

2 points - the presence in the facies, in addition to the above elements, of a dendritic component;

3 points - most or all of the field of vision is formed by large dendritic structures, amorphous bodies and single crystals are few.

2. Crystallizability. It is determined on a 4-point scale:

0 points - no crystallization centers;

1 point - single small crystals of a predominantly single row (not more than 10 in the field of view with a total magnification of the microscope × 56);

2 points - a significant number of crystallization centers (more than 10 in the field of view with a total magnification of the microscope × 56);

3 points - the entire field of view is covered with crystals of a single crystalline or dendritic row.

3. Type of interaction of crystalline and amorphous structures. There are two options for implementing the indicator:

a) sticking of amorphous particles to large crystalline formations;

b) the displacement of elements of an amorphous structure from crystalline structures.

4. The degree of destruction of the facies. It is expressed in four degrees:

0 degree - all elements of the facies of the correct configuration, are not destroyed as a whole, or in separate areas; there are no signs of destruction of the texture of the facies;

I degree - facies elements have initial signs of destruction, destructive changes in texture are not observed;

II degree - numerous destroyed or altered structures are visualized; there are local violations of texture integrity;

III degree - all elements of the facies are destroyed, it is impossible to distinguish the individual parts of the facies and structure, the sample is a shapeless mass of amorphous, often colored, material; There are clear signs of texture damage.

5. Uniform distribution of crystalline and amorphous elements of the facies. Interpreted using a six-point scale:

0 points - complete randomness of the facies, the presence of heterogeneous elements, voids, places of accumulation of crystalline structures, different orientation of formations in the field of view.

1 point - moderate grouping of crystals, single sections of their correct construction are outlined, occupying less than 30% of the total area of the field of view, the direction of the figures is still chaotic.

2 points - there are clear “islands” of ordering, occupying from 30 to 50% of the field of view (at least three are studied), distances between elements in groups are approximately equalized, some regularity of the direction of structural formations of the facies is recorded.

3 points - elements of the facies in a significant number (more than 50% of the total) are structured, islands of uniformity pass into areas of a relatively large area. Inside these zones, the correct location and uniformity of the distances between the individual formations are observed. The regularity of the direction of the elements and zonality are visible quite clearly.

4 points - facies elements are mostly structured (over 75% of the total), the rest are distributed by islands in the field of view, more often in the marginal zone. The distances between the individual formations are almost constant. The orientation of the elements obeys a certain regularity over almost the entire field of view.

5 points - all elements of the tezigraphic facies are clearly structured throughout the field of view, which is confirmed by the consideration of several fields. The division into the central, intermediate and marginal zones is clearly visible. The boundaries of the latter can be set even without a microscope. The distances between the elements of the picture are constant, the orientation of the figures is correct, regular on the entire surface of the facies.

6. The severity of the facies cellularity. The assessment is made on a six-point scale:

0 points - the complete absence of signs of the appearance of cellularity, the uniformity of the picture, there is no allocation of "islands" of crystals. Facies represent a single “layer” of crystalline formations.

1 point - the presence of the first signs of heterogeneity, "crushing" of the crystalloscopic picture:

- the beginning of the separation of groups of elements (less than 30% of the number of all entities that occupy less than 30% of the field of view);

- the appearance of a certain heterogeneity of the picture;

- the beginning of the "crushing" of a single "layer" of crystalline figures.

2 points - there is a fairly visible tendency to “crush” facies, the formation of “islands” of crystals:

- the number of isolated elements in the "islands" - from 30 to 50% of the number of all structures, they occupy more than 30% of the field of view of the facies;

- pronounced heterogeneity, zoning of the picture;

- the process of “dividing the facies into sections is visualized, the forming crystallization belts are recorded, which serve as the boundaries of the latter, in some cases not along the entire length, 1 crystal thick.

3 points - pronounced changes in the facies are observed:

- the elements in the "islands" make up from 50 to 75% of the total number, the surface they occupy is more than 50% of the field of view (across several fields);

- pronounced heterogeneity, "graininess" of the picture;

- clearly visible process of "dismemberment" of the facies in several fields of view;

- crystallization belts are quite distinct, formed by more than one row of crystalline structures.

4 points - signs of the appearance of cellularity are reliably visible:

- the number of structures grouped into cells from 75 to 100% of the total;

- grouped elements occupy the entire field of view (when studying at least three fields);

- “fragmentation”, “graininess” of the picture are very clearly expressed;

- crystallization belts are formed by more than one row of crystals, are present on the entire surface of the field of view, completely surround the "islands".

5 points - the picture is characterized by the following most diagnostically significant morphological characters:

- the number of structures grouped into cells from 75 to 100% of the total;

- grouped elements occupy the entire field of view (when studying several, at least three fields of view);

- very clearly expressed "fragmentation", "graininess" of the picture;

- crystallization belts are formed by more than one row of crystals, are present on the entire surface of the field of view, completely surround the "islands";

- there are “faults” in the picture (except for serum facies, for which this phenomenon is an independent diagnostic sign).

7. The severity of individual crystallization zones. It is expressed in points (from 0 to 3):

0 points - the impossibility of distinguishing zones of crystalloscopic facies;

1 point - with a small increase in the light microscope, the allocation of zones is not clear, slight signs of zoning are observed;

2 points - the presence of facies zones is clearly determined with a small magnification of the microscope;

3 points - clear visualization of areas even without a microscope.

8. The severity of the regional protein zone. This indicator is evaluated on a 4-point scale:

0 points - the complete absence of the edge zone, it is impossible to highlight the edge zone under the "small" (smaller × 56) magnification of the microscope;

1 point - the edge zone is present indistinctly and / or not throughout the facies;

2 points - with microscopy along the entire perimeter of the micropreparation, the presence of a pronounced edge zone is noted;

3 points - the ratio "the width of the edge zone: the radius of the facies" exceeds 1: 3;

4 points - pronounced edge zone in combination with the presence of "faults";

5 points - the edge zone is visualized even without the use of a microscope.

9. Relief texture. Estimated by a direct 4-point scheme:

0 points - no signs of texture;

1 point - texture elements (sections) barely noticeable under microscopy;

2 points - facies texture that is clearly distinguishable under microscopy;

3 points — micropreparation texture visualized without using a microscope.

This method does not require specialized equipment and additional reagents, except for a microscope with an increase of not more than × 56 and slides, both for the preparation procedure of the preparations and for the study itself, it takes 10-15 minutes, is simple to perform, and economical.

A method for the diagnosis of burn toxemia is as follows.

On a previously defatted, washed and dried glass slide apply 3 samples of patient blood serum with a volume of 100-200 μl. Subsequently, to obtain the biosubstrate facies, the micropreparation is dehydrated in a stream of warm air (temperature 40-50 ° С, humidity 20-30%). In this case, the horizontal position of the glass and the corresponding direction of flow ensures dehydration of the samples under the same conditions, preventing their unification. The drying time is 15-20 minutes.

Then, using a high-resolution digital camera, a digital image of the preparation is obtained, which is transferred to the computer's memory and the basic structure-forming elements of the facies are studied.

Based on the calculation of indicators form a pattern of their values. If it matches the burn toxemia pattern (Table 1), its presence is judged, for which the necessary signs are the coincidence of at least six out of nine parameters. With such an assessment, the sensitivity of the analysis is 86%, the specificity is 78% with respect to the "gold standard" - the method for determining medium molecular peptides (Malakhova M.Ya., 1995). Reducing the number of matches to five is not diagnostically significant, because in this case, endotoxemia is laboratory confirmed in less than 25% of cases, which does not allow to recognize this level as acceptable.

Clinical example.

Patient Z., 46 years old, was admitted to the department with a diagnosis of a burn with a flame of a voltaic arc and a flame from tanned clothes of I-II-IIIAB-IV degrees of the face, trunk, limbs on an area of 30% of the body surface. Burn shock. At the time of blood collection, the patient's condition is severe, a burn disease in the stage of acute toxemia. The patient is lethargic, complains of general weakness and nausea. Hyperthermia up to 38 ° C is noted. In the general analysis of blood: leukocytosis up to 22 · 10 ⁹ / l, ESR up to 20 mm / h. In biochemical analysis: hypoproteinemia (50 g / l) due to a decrease in all protein fractions (albumin 30 g / l, globulins 20 g / l), hyperbilirubinemia (22 μmol / l), hypercreatininemia (139.5 μmol / l), level urea 9.3 mmol / L; SMP E ₂₅₄ - 0.45; E ₂₈₀ - 0.54; myoglobin titer 1:64. These indicators indicate the presence of a moderate degree of endotoxemia in a patient. The patient received a course of antibiotics, painkillers, and infusion-transfusion therapy was carried out.

When conducting crystalloscopic studies (table 2, figure 2), the patient's blood serum facies was characterized as follows: structural index - 1.5 points; crystallizability - 2.5 points; type of interaction of crystals and amorphous bodies - sticking; the degree of destruction of the facies - 1.5 points; uniform distribution of crystalline and amorphous elements of the facies - 3 points; the degree of severity of the facies cellularity - 2 points; the severity of individual crystallization zones - 1 point; the severity of the regional protein zone - 1 point; texture relief - 2 points. According to the results of crystalloscopy, 8 out of 9 indicators are within the burn endotoxemia pattern, which is confirmed by other verification methods.

table 2 The results of crystalloscopy of blood serum of patient Z. Indicator SK patient Z. "Pattern" Coincidence Structural index 1,5 1-2 + Crystallizability 2.5 2-3 + Type of interaction of crystals and amorphous bodies sticking sticking + The degree of destruction of the facies 1,5 1-2 + Uniform distribution of sample elements 3 2-3 + The severity of cellularity 2 3-4 - Facialization one 0-1 + The severity of the marginal zone one 1-2 + Sharpness texture sample 2 2-3 + Conclusion: the presence of 8 matches allows to indicate the presence of burn endotoxemia

Figure 1 shows a crystalloscopic picture of the blood serum of a healthy person. A wide marginal zone (1), clear regular centripetal faults (2), signs of moderate crystallization in the form of single elements (3) are visible. Figure 2 presents the crystallogram of the blood serum of patient 3, which shows a narrow marginal zone (1), irregular, randomly located faults (2) and pronounced destruction of the crystalline elements of the facies (3).

Claims (1)

A method for the diagnosis of burn toxemia, consisting in the study of blood serum, characterized in that the blood serum of the patient in a volume of 100-200 μl is applied to a glass slide and the drug is dehydrated in a stream of warm air at a temperature of 40-50 ° C and a humidity of 20-30% in for 15-20 minutes in a horizontal position, the obtained crystalloscopic facies is compared with the burn toxemia pattern according to the following indicators: structural index, crystallizability, type of interaction of crystalline and amorphous structures, degree of facies destruction, uniform distribution of crystalline and amorphous elements of the facies, degree of severity of the facies cellularity, severity of individual crystallization zones, severity of the edge protein zone and texture embossment, and if at least six out of nine criteria values match the pattern, burn toxemia is diagnosed.

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