RU2627650C1 - Method for wound process course prediction - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method for wound process course prediction includes microscopic examination of the textures obtained as a result of studied biological fluid drying on a degreased slide in the form of a droplet at a temperature of 20-30C°, the study is carried out on isotropic textures, and blood serum is used as the biological fluid, which is dried for 24 hours, and then microscopically examined in transmitted light and if isotropic textures in the form of lingual fields, convex rounded formations, three-beam cracks are revealed, a prediction is made on the further course of the wound process - presence of one of isotropic textures types or their combination indicates a complication of wound healing.

EFFECT: method allows to identify the features of biomaterial physico-chemical structuring in case of pathology, it is simple in execution, feasible in a short time up to 24 hours and available in any clinical diagnostic laboratory.

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Description

The invention relates to medicine, in particular to a laboratory research method, and can be used to predict the course of the wound process.

The wound process is a complex set of general and local reactions of the body in response to the wound, providing wound healing.

In uncomplicated cases, the wound process proceeds in 3 phases: alteration, exudation and proliferation.

The initial stages of the wound healing process (1-4 days after injury) are characterized by the excitation of the sympathetic part of the autonomic nervous system, accompanied by an increased release of adrenaline into the blood, under the influence of which the body's vital activity increases, the basal metabolism, the breakdown of proteins, fats and glycogen increases, and permeability decreases cell membranes, regeneration mechanisms are suppressed, blood coagulation increases. The activity of the adrenal cortex increases, releasing glucocorticoid hormones, which have an anti-inflammatory effect. Thus, in response to the resulting injury, an adaptation syndrome develops.

The further course of the wound process is characterized (4-10th day after injury) by the increased influence of the parasympathetic department of the autonomic nervous system, the action of mineralocorticoid hormones, aldosterone, and the regeneration process is activated. In this phase, the metabolism, especially protein metabolism, normalizes, the wound healing process is activated.

A known method for predicting the course of the wound healing process with purulent-inflammatory diseases of the maxillofacial region of microbial etiology (RU No. 2405456, АВВ 10/00, 10.12.2010), the essence of this method is to study the microbiological picture of the wound discharge. In this case, the presence or absence of microorganisms and their concentration are determined, and in an immunological study, the total number of leukocytes in peripheral blood, ESR, the levels of average circulating immune complexes (CEC), antibodies to streptococcus are recorded.

The disadvantage of this method is that it takes a lot of time to get the results of the study, the method is expensive, technically more complicated, the forecasting result is formed by the time the treatment and diagnostic measures are completed.

A known method for predicting the course of the wound healing process (RU No. 2285924, G01N 336/53, 20.10.2006), the essence of which is that the content of proinflammatory interleukins TNFα, IL-1α, anti-inflammatory interleukins IL-4, IL-10 is determined in blood serum . In patients, the integral indicators of pro-inflammatory and anti-inflammatory interleukins are calculated, then the ratio of their content in the patient to the average value in healthy individuals is calculated. Then calculate the prognostic index of inflammation (PIV).

The disadvantage of this method is that it is expensive, and the calculation of the index is conditional, and, accordingly, the forecast is relatively inaccurate.

A known method for predicting the course of the wound process during deep frostbite of the lower extremities (RU No. 2285928 G01N 33/66, 10.20.2006), the essence of which is that they perform a microscopic examination of blood flow in the affected limb using laser Doppler flowmetry, calculate hemodynamics and indirectly determine the further course of the wound process.

The disadvantage of this method is that in the absence of blood flow in the tissues (deep ischemia) it is impossible to predict the further course of the wound process, and the study itself is highly costly.

A known method for predicting the outcome of the course of the wound healing process (RU No. 2142635, G01N 33/48, 10.12.1999), the essence of which is that they conduct a microscopic examination of the anisotropic textures of dried biological material from a detachable wound in an amount of 10-100 μl, while mixing with 0.1-0.25 ml of physiological solution of sodium chloride and dried in the form of a drop on an open surface at 20-30 ° C. This method is selected by the authors as a prototype.

The disadvantage of this method is that the proteins become more diluted when mixed with physiological saline, the introduction of a crystal-forming solution makes it difficult to interpret the results compared to using native serum. The concentration of protein in the wound discharge is significantly lower compared to that of blood serum, and therefore this method is difficult to reproduce. The difficulty also lies in the fact that during the wound process, the discharge occurs, as a rule, in the exudation stage, while at the other stages of the wound process this biological fluid is absent, which makes it impossible to predict the complications of wound healing. In addition, the disadvantage of this method is the duration of the forecast, about 72 hours, which makes the forecast irrelevant by the time the diagnostic information is received.

The objective of the invention is the development of criteria for predicting the course of the wound healing process based on the analysis of changes in the texture of dried blood serum.

The technical result achieved by the invention is the speed of obtaining diagnostic information, the ease of use of the method, the ability to predict the course of the wound process at the molecular level at various stages of wound healing.

The technical result is achieved by the fact that a microscopic study of the textures obtained by drying the test biological fluid on a defatted glass slide in the form of a drop at a temperature of 20-30 ° C is carried out, the study being carried out on isotropic textures, and blood serum is used as biological fluid, which is dried for 24 hours, and then microscopic examination is carried out in transmitted light, and, if isotropic textures in the form of language fields are detected, round rounded formations, three-beam cracks, make a prediction on the further course of the wound process, namely: in the presence of one of the species or their combination indicates a complication of wound healing.

The essence of the method is to identify patterns of intermolecular interaction in the protein-salt system when a drop of a blood serum is dried and to predict wound healing by the presence or absence of isotropic textures.

The isotropic textures identified by the authors in the form of language fields, convex rounded formations, and three-beam cracks are universal for typical pathophysiological processes, however, they were not previously used to predict the course of the wound process.

To date, it is known that the appearance of isotropic textures such as “language fields” is a sign of the inflammatory process (Shabalin V.N. et al. Morphology of the liquid media of the eye (a new theory of involutive cataractogenesis): monograph. - M .: Medicine, 2004. S. 157-160). This isotropic texture is pathognomonic for the active phase of the inflammatory process and indicates ongoing intermolecular disorders (Fig. 1).

The appearance of round convex isotropic inclusions of the type “toxic plaques” (Shatokhina SN, Shabalin VN Atlas of structures of non-cellular human tissues in norm and pathology in 3 volumes. Volume II. Morphological structures of blood serum. - M. - Tver: LLC Triada Publishing House, 2013. S. 240-245) characterize the depth of biochemical shifts and the severity of protein disorders, medium-weight molecules, which can be interpreted as the degree of endogenous intoxication of the body (Fig. 2).

The appearance of isotropic textures of the “three-beam cracks” type in the blood serum facies (Shabalin V.N. et al. Morphology of the eye fluid (new theory of involutive cataractogenesis): monograph. - M .: Medicine, 2004. P. 158-159) about stagnation in tissues due to changes in tissue perfusion parameters (Fig. 3).

In practice, the method is as follows.

A patient is sampled from v. intermedia cubiti in a volume of up to 5 ml, then centrifuged in a test tube without preservatives from 2000 to 2500 rpm for 15-20 minutes, then apply 20 μl of blood serum in the form of a drop on the surface of a glass slide. After 24 hours of exposure to blood serum at 20-30 ° C, a microscopic examination is carried out at a multiple of × 25 to × 100 in transmitted light and the presence or absence of isotropic textures is determined in it. The conclusion is made: about a positive forecast in the absence of isotropic textures, about a negative forecast (complicated flow) in the presence of isotropic textures.

This method allows you to identify the characteristics of the physico-chemical structuring of the biomaterial in pathology, simple to implement, feasible in a short time up to 24 hours and is available in any clinical diagnostic laboratory.

Clinical example 1

Patient B., 62 years old, was operated on in the surgical department for a restrained postoperative ventral hernia.

The results of the study of the morphological picture of blood serum in the first day after the operation: the presence of isotropic textures in the form of three-beam cracks, as well as language fields, which indicated an unfavorable course of the wound process.

Antibiotic therapy was prescribed. When re-examining blood serum on the 5th day after surgery, such isotropic textures were not detected. The wound healed by first intention, the sutures were removed on day 12. The patient was discharged in satisfactory condition 12 days after surgery.

Clinical example 2

Patient N., 34 years old, was operated on in the surgical department for a restrained left-sided oblique inguinal hernia.

The results of the study of the morphological picture of blood serum in the first day after the operation: the absence of isotropic textures, which indicated a favorable course of the wound process.

An additional correction of pharmacotherapy was not required. A repeated examination of blood serum on the 5th day after the operation also did not reveal isotropic textures. The wound healed by first intention, the sutures were removed on day 7. The patient was discharged in satisfactory condition on the 7th day after surgery.

Claims (1)

A method for predicting the course of the wound healing process, consisting in a microscopic study of textures obtained by drying the test biological fluid on a fat-free glass slide in the form of a drop at a temperature of 20-30 ° C, characterized in that the study is carried out on isotropic textures, and is used as a biological fluid blood serum, which is dried for 24 hours, and then carry out microscopic examination in transmitted light and when detecting isotropic textures in the form of tongues fields, convex circular formations, three-prong cracks do forecast the further course of the wound process - the presence of one of the types of isotropic texture or a combination of evidence of wound healing complications.

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