RU2754836C1 - Method for diagnosing volumetric formations of the brain - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to medicine, namely to laboratory research methods and relates to the differential diagnosis of benign brain tumors and malignant tumors. The cerebrospinal fluid is examined by the method for edge dehydration in polarized light, while the cerebrospinal fluid is preliminarily centrifuged for 30 minutes at 10,000 rpm, then the supernatant sample is placed in a water bath at + 42°C for 3-5 minutes and dehydrated in within 3-5 days. Then the analytical cells are microscoped and when single alien anisotropic structures outside the dendrites are detected in the visual fields or when small clusters of foreign anisotropic structures with inclusions in dendrites and outside the dendrites are detected, a benign brain formation is diagnosed. If a dense ring of foreign anisotropic structures is found around the dendrites and with inclusions in them, a malignant tumor is diagnosed. The proposed method allows with high reliability to differentiate benign or malignant volumetric formation of the brain at the preoperative stage.

EFFECT: method is not laborious, does not require additional use of expensive drugs, is highly economical and can be widely used in neurosurgical practice to assess the prognosis of the course of the disease, select the optimal tactics and treatment method for patients with masses of the brain, aimed at improving the quality of life and increasing its duration.

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Description

The invention relates to medicine, namely, to laboratory research methods and relates to the differential diagnosis of benign brain tumors and malignant tumors.

The diagnosis of masses, due to the localization of the tumor inside the skull, is difficult. The diagnosis of an oncological disease is finally made only after a histological conclusion, that is, without a histological or cytological examination, the diagnosis is incompetent. Therefore, the relevance of differential diagnosis of benign tumors and malignant brain tumors at the preoperative stage is beyond doubt, since it allows assessing the prognosis of the course of the disease, choosing the optimal tactics and method of treatment aimed at increasing the life expectancy of patients and its quality.

A known method for the diagnosis of benign and malignant glial tumors (RF Patent 2154830, IPC G01N 33 / 53.50, publ. 2000) by examining the patient's blood serum, in which a passive hemagglutination reaction is performed with benign, anaplastic and glioblastoma erythrocyte diagnostics with corresponding polyantigenic tissue preparations glial neoplasms. In the presence of positive results with a benign and anaplastic diagnosticum or only with a benign one, benign glial tumors are diagnosed, and in the presence of positive reactions with all diagnosticums, malignant tumors are diagnosed.

The disadvantage of this method is the high labor intensity of the study, the use of expensive reagents and the difficult interpretation of the results.

A known method for the differential diagnosis of brain tumors (RF Patent 2258932, IPC G01N 33/483, publ. 2005), including carrying out a crystallographic study of cerebrospinal fluid. When the protein concentration in CSF samples is from 0.50 g / l and above with a predominance of crystals in the form of oppressed dendrites in the crystallographic pattern, a malignant brain tumor is diagnosed, and when the protein concentration is below 0.50 g / l and the predominance of crystals in the form of branched dendrites is benign a brain tumor.

The disadvantage of this method is that the diagnosis of a benign or malignant brain tumor only by the predominance of one or another type of crystals of the same salt (sodium chloride) is not accurate enough, because acts as a subjective sign, which can lead to a serious error in the differential diagnosis of the type of volumetric formation of the brain.

The closest is a method for the diagnosis of benign and malignant brain tumors (Shabalin V.N., Shatokhina S.N. Morphology of human biological fluids. M .: Khrysostom, 2001. p. 244), including the study of cerebrospinal fluid by the method of edge dehydration in polarized light , identification of morphotypes in the sample in the form of dendrites and alien anisotropic structures.

However, this method does not provide a sufficiently high accuracy of differential diagnosis of masses of the brain, since there are artifacts in the cerebrospinal fluid that can either obscure the signs of atypia, or their localization can be regarded as a sign of malignancy of the masses.

The objective of the present invention is to eliminate these disadvantages and improve diagnostic accuracy by identifying the location of morphotypes and foreign anisotropic structures of cerebrospinal fluid.

To solve this problem, the cerebrospinal fluid is examined by the method of edge dehydration in polarized light and morphotypes in the form of dendrites and foreign anisotropic structures are identified in the sample. Before setting up the method of edge dehydration, the cerebrospinal fluid is centrifuged for 30 minutes at 10,000 rpm, then the supernatant sample is placed in a water bath at + 42 ° C for 3-5 minutes and dehydrated for 3-5 days. Then the analytical cells are microscoped and when single alien anisotropic structures outside the dendrites are detected in the visual fields or when small clusters of foreign anisotropic structures with inclusions in the dendrites and outside the dendrites are detected, a benign brain formation is diagnosed. If a dense ring of foreign anisotropic structures is found around the dendrites and with inclusions in them, a malignant tumor is diagnosed.

This approach to research - high-speed and long-term centrifugation makes it possible to precipitate cells and all destructive elements in the form of organized particles. That is, conditions are created for the formation of morphotypes directly from the cerebrospinal fluid, which, upon transition to the solid phase, provide diagnostic information at the molecular level.

Another new and important aspect of the study is the creation of conditions for more active microaggregation of structures, which is achieved by placing cerebrospinal fluid in a water bath for 3-5 minutes at 42 ° C.

The inclusion of these actions in the study allows one to avoid the influence of artifacts on the formation of morphotypes and to transfer to the macrolevel all the connecting elements characterizing both physiological structures and atypical metabolites of a malignant tumor. All this makes it possible to significantly improve the accuracy of differential diagnosis of masses.

FIG. 1 shows a dendrite surrounded by single alien anisotropic structures (x50); in fig. 2 shows a dendrite, in the center of which there is a small accumulation of alien anisotropic structures (arrow), the same small accumulations of alien anisotropic structures are determined outside the dendrite (arrows) (x200); in fig. 3 shows a dense ring of alien anisotropic structures located around the dendrites and included in their composition (x150).

The method is carried out as follows:

Cerebrospinal fluid is collected from a patient with a mass of the brain and delivered to the laboratory, where it is centrifuged at 10,000 rpm for 30 minutes. Then 0.5 ml is taken from the supernatant into a test tube and placed in a water bath for 3-5 minutes at 42 ° C. Then, at least three drops of 0.02 ml of cerebrospinal fluid are applied to the surface of the slide, cover them with cover glasses for dehydration for 3-5 days at a temperature of + 25 ° C and a relative humidity of 55-60%. Then microscopy of analytical cells is carried out in polarized light and when single alien anisotropic structures (see Fig. 1, example 1) or small clusters that are located outside the dendrites and are part of them (see Fig. 2, example 2) are diagnosed in the fields of view benign formation of the brain, and upon detection of dendrites surrounded by a dense ring of anisotropic structures and included in them (see Fig. 3, example 3), a malignant tumor is diagnosed.

Diagnostics according to the proposed method is shown in the following clinical examples.

Example 1

Patient K., 62 years old, was admitted to the Department of Neurosurgery with a clinical diagnosis: volumetric brain mass. Cerebrospinal fluid was taken from the patient at the preoperative stage and examined by the method of marginal dehydration according to the proposed method. In this case, after high-speed centrifugation, a sample of the supernatant was placed in a water bath at + 42 ° C for 5 minutes and dehydrated for 3 days. Microscopy of analytical cells revealed single alien anisotropic structures in the form of ovals, spheres, not included in dendrites (Fig. 1).

Conclusion: benign brain formation.

Histological diagnosis after surgery: benign brain tumor - fibrillar astrocytoma.

Example 2

Patient S., 57 years old, was in the department of neurosurgery with a clinical diagnosis: volumetric formation of the brain.

The patient received cerebrospinal fluid before the operation and examined by the method of marginal dehydration according to the proposed method. In this case, after high-speed centrifugation, a sample of the supernatant was placed in a water bath at + 42 ° C for 3 minutes and dehydrated for 4 days. Microscopy of analytical cells revealed dendrites, inside and outside of which small accumulations of small alien anisotropic structures in the form of petals, ovals, triangles were revealed (Fig. 2).

Conclusion: benign brain formation.

During the operation, a cerebral aneurysm was established.

Example 3

Patient A., 66 years old, was in the department of neurosurgery with a clinical diagnosis: volumetric formation of the brain.

The patient received cerebrospinal fluid before the operation and examined by the method of marginal dehydration according to the proposed method. In this case, after high-speed centrifugation, a sample of the supernatant was placed in a water bath at + 42 ° C for 4 minutes and dehydrated for 5 days. Microscopy of analytical cells revealed dendrites, which were surrounded by a dense ring of foreign anisotropic structures and entered the dendrites (Fig. 3).

Conclusion: a malignant brain tumor.

Histological diagnosis after surgery: malignant tumor - glioblastoma.

Thus, the proposed diagnostic method provides the clearest signs that make it possible to differentiate benign formations from malignant tumors at the preoperative stage of patient examination. Such information is always in demand by the doctor to determine the further tactics of patient management, the optimal methods of his treatment and prevention of possible complications in the course of treatment.

It should be noted that a change in the permeability of the blood-brain barrier during the development of volumetric formations of the brain leads to the destruction of its structures with the subsequent development of immune responses directed against brain and tumor antigens, as well as the elimination of brain and tumor-specific substances into the blood and cerebrospinal fluid. These structures can be visualized during the transfer of cerebrospinal fluid to the solid phase by the method of edge dehydration. Elements of tissue metabolites of the brain form specific formations with distinctive features indicating the severity of a particular pathology causing the destruction process.

In the neurosurgical department, using the proposed diagnostic method, 44 patients with masses of the brain were examined at the preoperative stage. In the study of the morphotypes of the cerebrospinal fluid of patients, 28 cases of benign tumors and 16 cases of malignant tumors were diagnosed. Further comprehensive examination of patients, including a histological report, confirmed a malignant tumor in all 16 patients. Among 28 patients with benign tumors, 7 were diagnosed with a cerebral aneurysm, and 21 patients with a benign tumor.

Thus, the proposed method allows with high reliability to differentiate benign or malignant volumetric formation of the brain at the preoperative stage. The method is not laborious, does not require additional use of expensive drugs, is highly economical and can be widely used in neurosurgical practice to assess the prognosis of the course of the disease, select the optimal tactics and treatment method for patients with masses of the brain, aimed at improving the quality of life and increasing its duration.

Claims (1)

A method for diagnosing volumetric formations of the brain, including the study of cerebrospinal fluid by the method of edge dehydration in polarized light, identifying morphotypes in the form of dendrites and foreign anisotropic structures in the sample, characterized in that the cerebrospinal fluid is pre-centrifuged for 30 minutes at 10,000 rpm, then the sample the supernatant liquid is placed in a water bath at + 42 ° C for 3-5 minutes, dehydrated for 3-5 days and when single alien anisotropic structures outside the dendrites are detected in the visual fields or small accumulations of foreign anisotropic structures with inclusions in dendrites and outside the dendrites are detected diagnose a benign brain formation; when a dense ring of foreign anisotropic structures is found around the dendrites and with inclusions in them, a malignant tumor is diagnosed.

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