RU2673659C1 - Method of diagnostics of malignant neoplasms of the brain - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to the field of medicine, namely to diagnosis by examining the physical and chemical properties of blood, and can be used in clinical practice for the diagnosis of malignant neoplasms of the brain. Method includes the examination of blood plasma by instrumental methods and diagnosis. Investigate the free radical activity and the content of myoglobin in blood plasma, erythrocyte catalase, whole blood acetylcholinesterase. With an increase in free radical activity of blood plasma by more than 500 % of the physiological norm of 1.25±0.12 mV and an increase in the content of myoglobin by more than 25 % of the physiological norm of myoglobin in blood plasma 22.63±6.33 ng/ml, while reducing the activity of erythrocyte catalase by more than 33 % of the physiological norm 15.24±3.21 mmol/L and a decrease in the activity of whole blood acetylcholinesterase by more than 35 % of the physiological norm 24.36±5.72 mg/ml/hour/0.1 g Hb diagnose a malignant neoplasm of the brain.

EFFECT: invention provides early diagnosis of malignant neoplasms of the brain, without reducing its accuracy, specificity and objectivity, providing screening diagnostics for a large population of people, simplifying the method, reducing examination time and cost.

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Description

The invention relates to medicine, namely to diagnosis by examining the physical and chemical properties of blood and can be used in clinical practice for the diagnosis of malignant neoplasms of the brain.

To date, the prevalence of brain tumors is from 5 to 14 cases per 100 thousand people (Matsko D.E. Neurosurgical pathology. Guide. - St. Petersburg: B.I., 2012. - 405 p.). Around the world, about 240,000 new cases and about 175,000 deaths from brain tumors are recorded annually. Glial tumors are among the most common neoplasms of the brain and make up, on average, from 50 to 55% of all primary cerebral neoplasms (Zozulya Yu.A. Gliomas of the brain. - Kiev: UIPK "Exob", 2007. - 632 p.) . Most of them are malignant gliomas with a high degree of anaplasia — grade III-IV, and make up 55-60% of all primary glial tumors (Olyushin V.E. Glial brain tumors: a brief review of the literature and the protocol for treating patients // Neurosurgery. - 2005. - No. 4. - S. 41-47.). Early, reliable information about the degree of malignancy of the tumor will optimize the treatment process and significantly increase its effectiveness.

A known method for diagnosing the degree of anaplasia of malignant and benign brain tumors using proton magnetic resonance spectroscopy (PMRS) (Trufanov G.E., Tyutin L.A. Magnetic resonance spectroscopy. - SPb .: EL-BI-SPb, 2008. - 239 p.).

The method is based on determining the content of certain metabolites in tumor tissues and in the peritumoral zone. Choline, lactate, creatine, phosphocreatinin, myo-inosine, glutamate, glutamine, lipids, N-acetyl aspartate and their ratios are most often investigated.

The known method involves obtaining a spectrum from a cubic section (voxel) of brain substance or tumor 2 × 2 × 2 cm (8 cm ³ ) in size and analyzing the frequencies in the spectrum recorded from this voxel, resulting in the distribution of certain metabolites in the tissue on a chemical shift scale (rt). The ratio between the peaks of metabolites in the spectrum, a decrease or increase in the height of individual peaks of the spectrum, allows non-invasive assessment of biochemical processes occurring in tissues. Malignant tumors are characterized by a low ratio of N-acetyl aspartate / creatine, an increase in the ratio of choline / creatine and, in some cases, the appearance of a peak lactate.

The technical result of the known method is to enable the assessment of various volumetric formations of the brain and differential diagnosis of astrocytomas, ependymomas and primitive neuroepithelial tumors, presumably determining the type of tumor tissue. The known method allows you to diagnose with a high degree of probability the degree of malignancy of the process.

However, the disadvantages of this method include the lack of diagnostic accuracy, since only one section of a small volume of tissue (voxel) is examined. With multivoxel spectroscopy, the study is carried out only in one plane on a relatively small area of the neoplasm. However, it is known that a tumor is heterogeneous in its structure and activity of tumor cells. And to obtain information about the activity of the entire tumor using this method is not possible. In addition, the location near the study area of bone structures, cerebrospinal fluid spaces, large vessels and foci of hemorrhage leads to the appearance of artifacts that distort the results of studies (Podoprigora A.E. Proton magnetic resonance spectroscopy in the diagnosis of volumetric brain diseases: abstract. Diss. ... Candidate of Medical Sciences. - M., 2002 .-- 30 p.). In addition, similar changes in metabolites are detected with abscesses, with pilocytic astrocytomas, which does not allow using PMRS to clearly differentiate the degree of anaplasia, malignant and benign brain tumors (Kornienko, B.N., Pronin I.N. Diagnostic. - M .: IE “Andreeva T.M.”, 2006. - 1327 p.). In addition, the study is contraindicated to be performed with the built-in pacemaker, vascular clamps, metal implants, cochlert prosthesis of the inner ear, pregnancy, claustrophobia, and the patient’s general general condition. Sometimes an anesthesia is necessary for conducting PMRS (for young children, adult patients with mental disorders or with pain), since during the procedure, the subject should be absolutely motionless for a long time.

For the prototype of the invention, a well-known method for the diagnosis of malignant neoplasms of the brain is selected, including the study of biological material using instrumental methods and diagnosis (Koshkin F.A., Chistyakov D.A., Nikitin A.G., Konovalov A.N., Potapov A.A., Usachev D.Yu., Pitskhelauri D.I., Kobyakov G.L., Shishkina L.V., Chekhonin V.P. Study of the profile of microRNA expression in brain tumors of different degrees of malignancy // Bulletin of Experimental Biology and medicine - 2014 - T. 157. - No 6. - S. 794-797.)

The known method is as follows.

Total RNA was isolated from blood plasma using the TriReagent reagent (MRC, USA). The RNA pellet was dissolved in 12 μl of RNA eluent (Interlabservis, Moscow). Complementary DNA (cDNA) is prepared using the StemLoop technology using the OT-1 reverse transcription kit (Synthol, Moscow). The reaction is carried out separately with primers for the microRNA-21 gene and the reference U6 gene (small nuclear RNA). The reaction mixture contains 5 μl of reaction buffer (2.5 X Reaction mixture), 10 pMol of a specific reverse transcription primer (Table 1), 0.5 μl of an RNase inhibitor, 0.5 μl of the MMLV enzyme (Synthol, Moscow) and 5 μl RNA solution. The reaction is carried out in a DT-Light thermocycler (DNA Technologies, Moscow) according to the following program: 16 ° С - 30 min, 42 ° С - 30 min, 85 ° С - 5 min, followed by cooling of the mixture to 4 ° С. The microRNA-21 gene expression study was carried out using real-time polymerase chain reaction (PCR) using the 2.5x PCR-PB reaction mixture in the presence of EVA Green dye (Synthol, Moscow) in a final volume of 25 μl. The PCR mixture contains: 10 μl of 2.5x PCR buffer B (with SynTaq DNA polymerase), 10 pM mol of forward and reverse primers and 5 μl of the corresponding cDNA sample. Amplification is carried out on a DT-Lite instrument (DNA Technologies, Moscow) taking into account the level of fluorescence through the FAM channel (470/525 nm). The amplification program is as follows: 95 ° С - 5 min, then 45 cycles (95 ° С - 15 sec and 60 ° С - 1 minute). Calculation of the relative level of gene expression of miRNA-21 is carried out according to the standard method ΔΔCt. Evaluation of the expression level of miRNA-21 by a semi-quantitative method is based on its comparison with the expression of the small nuclear RNA gene (snRNA) U6, which is used as a control due to the stability of its expression level. Signal intensity is expressed in relative fluorescence units (OEF). For each sample, two accumulation curves of the fluorescent signal of miRNA-21 and sniRNA U6 are built. The result of PCR-RV is the Ct value (threshold cycle) - the number of cycles required for the fluorescent signal to cross the threshold (background level). The Ct level is inversely proportional to the amount of the target nucleic acid in the sample (the lower the Ct level, the higher the number of target nucleic acid in the sample). With a more than twofold increase in the level of expression of micro-RNA-21 (compared with practically healthy people), a III-IV degree of malignancy of a brain tumor is diagnosed.

The known method allows to increase the sensitivity and specificity of detecting malignant forms of brain tumors and obtain reliable results that reflect the activity of invasive growth and the infiltrative process of this neoplasm.

The disadvantages of the method include its complexity and the complexity of the implementation. The known method is limited in application due to the lack of practical healthcare equipment in hospitals for its implementation. Another significant drawback of the method is the possibility of its use only for gliomas, and not for all brain tumors.

The task of the invention is to provide a method devoid of the disadvantages of the prototype.

The technical result of the invention is to provide early diagnosis, without reducing its accuracy, specificity and objectivity, providing the possibility of screening diagnostics of a large population of people, simplifying the method, reducing research time and cost.

The technical result is achieved by the fact that in the known method for the diagnosis of malignant neoplasms of the brain, including the study of blood plasma by instrumental methods and diagnosis, the free radical activity and content of blood plasma myoglobin, erythrocyte catalase, whole blood acetylcholinesterase, and with an increase in free radical activity of blood plasma more than by 500% of the physiological norm of 1.25 ± 0.12 mV, and an increase in the content of myoglobin by more than 25% of physiological blood norm of myoglobin in blood plasma is 22.63 ± 6.33 ng / ml, with a simultaneous decrease in the activity of erythrocyte catalase by more than 33% of the physiological norm of 15.24 ± 3.21 mM / l and a decrease in the activity of whole blood acetylcholinesterase by more than 35 % of the physiological norm 24.36 ± 5.72 mg / ml / hour / 0.1 g HB diagnose a malignant neoplasm of the brain.

The present invention meets the criterion of "novelty", since when conducting patent information research, sources of scientific, technical and patent literature that would discredit the novelty of the invention were not identified.

It is known from the prior art that a violation of free radical activity in malignant neoplasms is of a diagnostic nature and an increase in the level of free radicals is one of the triggers of carcinogenesis (Voleikov V.L.The beneficial role of reactive oxygen species // Biochemistry, 2004. No. 1, pp. 27-38 .). The brain is particularly vulnerable to free radicals because of its high oxygen consumption and high concentrations of readily oxidized polyunsaturated fatty acids. Therefore, the brain is most susceptible to oxidative damage.

Myoglobin is a multifunctional hemprotein, normally expressed mainly in myocytes. The main function of this protein is reversible binding to oxygen, storage and transport of it from hemoglobin. However, according to recent literature data, myoglobin is also detected in non-muscle tissues that lack oxygen (Kanatous S., MammenP., 2010). So, Flonta et al. (2009) showed in their works a high level of myoglobin in malignant tumors of human epithelial tissues (breast, ovary, intestine cancer) already at the very early stages of the disease. In this case, the tumor-suppressing role of myoglobin due to inhibition of mitochondrial activity is discussed.

Catalase is a heme-containing enzyme with a molecular mass of about 250 kDa that destroys hydrogen peroxide without the participation of oxygen acceptors, and hydrogen peroxide itself serves as an electron donor. The structure of the enzyme is a tetramer containing one tightly bound heme prosthetic group per subunit. The function of the enzyme is to prevent the accumulation of hydrogen peroxide in the cells, which is formed, in particular, during the dismutation of the superoxide anion and during the aerobic oxidation of reduced flavoproteins. Catalase refers to enzymes that retain their highest activity for the longest time, almost do not require activation energy, and the reaction rate catalyzed by this enzyme limits only the diffusion rate of the substrate to the active center of catalase (Pavlova R.N., Golovanova N.E. et al. Study of adaptogenic and antioxidant properties of Transfer Factor Advance // St. Petersburg State Medical Academy named after II Mechnikov, St. Petersburg, 2009).

Е, Montenegro MF, Cabezas-Herrera J. et al. Cholinergic system and cell proliferation. Chem. Biol. Interact. 2016; 259(Pt B): 257-265). Ацетилхолинэстераза выполняет роль опухолевого супрессора в культурах клеток опухолей даже при отсутствии ферментативной активности и может влиять на апоптоз раковых клеток негидролитическим путем, экспрессия и активность холинэстераз в тканях онкологических больных коррелирует с выживаемостью больных и связана со степенью злокачественности опухоли (Jin Q.H., Не H.Y., Shi Y.F., Lu Н., Zhang X.J. Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK. Acta Pharmacol. Sin. 2004; 25(8): 1013-1021).Recently, papers have appeared that discuss cholinesterase as potential tumor markers and the prospects of using changes in blood cholinesterase activity to predict the fate of patients after cancer (Moon J, Chun B. Utility of red blood cell acetylcholinesterase measurement in mechanically ventilated subjects after organophosphate poisoning. Respir Care. 2014; 59 (9): 1360-1368). Separate data were obtained on the involvement of acetylcholinesterase in the control of proliferation, the transition of cells to apoptosis, the development of inflammatory reactions and the formation of immunity (Campoy FJ, Vidal CJ,

E, Montenegro MF, Cabezas-Herrera J. et al. Cholinergic system and cell proliferation. Chem. Biol. Interact 2016; 259 (Pt B): 257-265). Acetylcholinesterase plays the role of a tumor suppressor in tumor cell cultures even in the absence of enzymatic activity and can affect cancer cell apoptosis non-hydrolytically, the expression and activity of cholinesterase in the tissues of cancer patients correlates with patient survival and is associated with the degree of tumor malignancy (Jin QH, He HY, Shi YF, Lu N., Zhang XJ Overexpression of acetylcholinesterase inhibited cell proliferation and promoted apoptosis in NRK. Acta Pharmacol. Sin. 2004; 25 (8): 1013-1021).

However, in the scientific and medical literature, information about the possibility of diagnosing malignant neoplasms of the brain using an integrated analysis of the activity of catalase, acetylcholinesterase and the content of myoglobin and free radical activity of blood plasma is not detected, which allows us to conclude that the proposed invention meets the criterion of "inventive step".

The technical result of the proposed method consists in a comprehensive assessment of the physico-chemical parameters of the blood with the determination of marker proteins and free radical activity. The present invention when used allows you to:

- provide early diagnosis of malignant neoplasms at the prehospital stage;

- provide the possibility of screening diagnostics during professional examinations;

- in the preoperative period, provide a prognosis of a benign or malignant nature of the tumors;

- provide high accuracy diagnostic tests; monitoring during antitumor therapy;

- to ensure the identification of metostases in the postoperative period and

- significantly reduce the time and complexity of the method;

- reduce the economic costs of diagnostics, because, unlike the prototype, expensive equipment and reagents are not required.

The technical result is confirmed by clinical and laboratory tests. The work examined the blood of 12 patients with malignant brain tumors - glioblastomas, astrocytomas (Grade III-IV) - 6 women and 6 men in the age category from 39 to 53 years. 7 patients with benign brain tumors - meningiomas, astrocytomas (Grade I-II) - 3 women and 4 men aged 37 to 59 years, before treatment. Blood from 10 practically healthy people (5 men and 5 women in the age category from 35 to 65 years old) was used as control.

The results of the study are presented using graphic material.

In FIG. Figure 1 shows a diagram of the free-radical activity of blood plasma in brain tumors: with benign brain tumors, with malignant brain tumors, in practically healthy people (control). * - differences with control group indicators are significant (p <0.05)

In FIG. 2. shows a diagram of the content of myoglobin in blood plasma in brain tumors: with benign brain tumors, with malignant brain tumors, in healthy people (control)

In FIG. Figure 3 shows a diagram of the activity of erythrocyte catalase in brain tumors: with benign brain tumors, with malignant brain tumors, in healthy people (control). * - differences with control group indicators are significant (p <0.05)

In FIG. Figure 4 shows a diagram of the total activity of whole blood acetylcholinesterase in brain tumors: with benign brain tumors, with malignant brain tumors, in healthy people (control). * - differences with control group indicators are significant (p <0.05)

The free-radical activity of blood plasma significantly increased in malignant neoplasms of the brain compared with practically healthy people and people with benign neoplasms more than 15 and 2.5 times, respectively (Fig. 1).

The level of myoglobin in blood plasma in malignant brain tumors exceeds those in practically healthy people of the control group by 24%. Moreover, the amount of hemprotein in plasma with gliomas with a low degree of anaplasia and benign neoplasms does not significantly differ from normal indications (Fig. 2)

The activity of catalase in red blood cells was significantly lower in brain tumors compared with practically healthy people (Fig. 3). With malignant neoplasms of the brain, catalase activity decreased more significantly - more than 3 times than with benign tumors - 1.4 times.

The total activity of blood cholinesterase was significantly lower (29%) in patients with malignant brain tumors compared with practically healthy people (24.36 ± 5.72 mg / ml / hour / 0.1 g HB) (Fig. 4) .

The proposed method is as follows.

A 10 ml blood sample is taken from a cubital vein into a test tube with citrate as an anticoagulant. The determination of acetylcholinesterase activity is carried out in whole blood. 0.5 ml of blood is diluted with distilled water to a total volume of 10 ml (20-fold dilution). 1.5 ml of 1/15 M phosphate buffer pH = 7.8; 0.5 ml of water (in the control for reagents - 0.3 ml of water and 0.2 ml of a 1% solution of proserin) and 1 ml of blood diluted 20 times. The tubes are thermostated for 15 minutes at 25 ° C, then 1 ml of 0.4% acetylcholinhydrochloride is added to them and incubated again for 30 minutes. At the end, add 1 ml of 25% trichloroacetic acid, the contents are mixed, centrifuged for 15 minutes at 3000 rpm. 2 ml of a mixture of equal amounts of 2M hydroxylamine hydrochloride and 3.5 N sodium hydroxide, 1 ml of hydrochloric acid (1: 3) and 1 ml of 0.37M ferric chloride are added to 1 ml of a centrifugate. The optical density of the solutions is measured on a photoelectrocalorimeter, wavelength 490 nm; 5 mm cuvette against water. The calculation is made according to the formula.

The determination of the concentration of myoglobin of free radical activity is carried out in blood plasma. For this, whole blood is centrifuged at 3000 rpm for 10 minutes and the red blood cells and plasma are separated.

The determination of myoglobin is carried out by the erythrocyte method (to determine the dilution ratio) and immunoturbodimetric - for the final analysis of the concentration of myoglobin. 50 μl of a 0.9% sodium chloride solution are added to 24 wells of a polystyrene tablet. 50 μl of plasma is added to the first well, and 50 μl of the sample solution is successively transferred from the well to the well, resulting in a 1: 2 dilution; 1: 4; 1: 8 etc. The last well is a control, a blood sample is not added there. Then, 25 μl of resuspended suspension of formalin erythrocytes sensitized with anti-myoglobin antibodies is successively added to each well. Then the contents of the wells are mixed and allowed to incubate for 30 minutes. The results are taken into account on the basis of a visual assessment of the presence of hemagglutination depending on the degree of dilution of the test sample using the table of correspondence of the amount of myoglobin and the titer value.

Having determined the concentration of myoglobin in a semi-quantitative way, assess the need for dilution of the sample and conduct an immunoturbidimetric test.

To do this, 60 μl of plasma is poured into a test tube and 1800 μl of reagent 1 containing 1.5% glycine solution in pH 8.3 buffer is added, after which it is thoroughly mixed and incubated in a water bath for 5 minutes. Then add 600 μl of reagent 2 containing latex particles coated with antibodies to myoglobin in a 1.5% glycine solution in pH 7.3 buffer, mix thoroughly and absorbance is measured for 30 seconds on a spectrophotometer at a wavelength of 580 nm.

Then incubated in a water bath for 5 minutes and again measured the optical density at a wavelength of 580 nm. The calculation of the difference in optical densities is performed according to the formula: delta D = D2-D1.

To calculate the concentration of myoglobin, a calibration curve is used. Free-radical activity of blood plasma is analyzed by the method of induced biochemiluminescence. To do this, 0.1 ml of blood plasma is added to the measuring cell; 0.4 ml of phosphate buffer (pH = 7.5) and 0.4 ml of iron sulfate solution (0.05 mmol). 0.2 ml of a 2% hydrogen peroxide solution is added to the cuvette installed in the cuvette holder of the biochemiluminometer, then the handle of the cuvette block is turned 180 degrees counterclockwise until it stops. The “Measurement” window appears on the display and the registration of the kinetic curve of the chemiluminescence signal begins. At the end of the set time (30 sec.), The measurement is interrupted and the necessary parameters of the chemiluminescent signal are calculated. The program goes to the processing window, on which the experience map appears, on which a normalized graph of the chemiluminescence kinetics and calculated signal parameters are shown (Imax, S, a, Z, tg1, tg2, Inc, Dec). In this paper, we consider the indicator Imax.

Catalase activity is determined in red blood cells. The red blood cells are washed by centrifugation at 3000 rpm with a 0.9% sodium chloride solution.

With washed red blood cells / homogenate, the following dilutions are obtained:

> I dilution - 0.2 ml of “washed” red blood cells / homogenate are poured into a test tube, 3.8 ml of Tris-acetate buffer (TAE) is added, pH = 8.0

> II dilution - 0.1 ml of I dilution and 4.9 ml of phosphate buffer are poured into a test tube, pH = 7.5

Next, make the following measurements:

> Pour into a cuvette for a spectrophotometer:

1. 1.0 ml of II dilution and 2.0 ml of phosphate buffer. After 60 seconds, the obtained value of the activity of catalase E1 is fixed.

2. 1.0 ml of pre-prepared hydrogen peroxide and 2.0 ml of phosphate buffer. The obtained numerical value of E2 should be in the range from 0.320 to 0.340 μatmg /.

3. 1.0 ml of I dilution, 1.0 ml of phosphate buffer and 1.0 ml of H2O2 - the result of spectrophotometry is the obtained value of the activity of catalase E3.

After these measurements, the value of catalase is calculated by the formula:

E = (E1-E3) * V * t * K - for blood catalase

E = (E1-E3) * V * t * Km - for catalase of brain tissue,

where E is the activity of catalase (μatmg /),

V is the volume of the applied sample (0.003 L), t is the incubation time (60 s), m is the mass of tissue in the applied sample (mg). K is the coefficient of millimolar extinction of hydrogen peroxide (22.2⋅103 mM-1⋅cm-1).

Conversion: 1 mmolmin / = 0.01667 mkat.

Free radical activity and plasma myoglobin activity, erythrocyte catalase content and whole blood acetylcholinesterase catalase are studied and with an increase in the free radical activity of blood plasma by more than 500% of the physiological norm of 1.25 ± 0.12 mV, and an increase in myoglobin content of more than 25% from the physiological norm of myoglobin in blood plasma 22.63 ± 6.33 ng / ml, while reducing the activity of erythrocyte catalase by more than 33% from the physiological norm of 15.24 ± 3.21 mM / l and a decrease in the activity of whole acetylcholinesterase vi more than 35% of the physiological norm 24.36 ± 5.72 mg / ml / hour / 0.1 g Hb diagnose malignant brain tumor.

Examples of specific performance are given in the form of extracts from the medical history

Example 1

Patient O., 43 years old. Was admitted to a neurosurgical clinic with a diagnosis of volumetric formation (glioma?) Of the left frontotoparietal-temporal lobe.

Complaints: weakness in the limbs on the right, rare bouts of epilepsy.

Anamnesis: sick for about a year. When cramps first appeared. Over the past month, weakness in the extremities has joined. After detecting a tumor on MRI, the patient is referred to a neurosurgeon.

Neurologically: consciousness is clear, oriented, critical, anisoreflexion D> S, Barre symptom on the right is positive, hemiparesis on the right is up to 4.5 points.

According to MRI with contrast enhancement from 12/07/16 a tumor of 9X5 cm in the posterior-frontal region with medial spread.

Upon admission to the hospital before the start of therapeutic measures, an analysis of the biochemical parameters of the blood of patient O., aged 43, was carried out: catalase activity = 10.98 mmol / l, acetylcholinesterase = 19.2 mg / ml / hour / 0.1 gNv, myoglobin concentration = 5 ng / ml and free radical activity Imax = 6.96 mV. In this way:

- the observed decrease in the activity of the red blood cell catalase of patient O. is equal to 27.95%, which is less than 33%;

- the observed decrease in the blood acetylcholinesterase activity of patient O. is 21.18%, which is less than 35%;

- the content of myoglobin in blood plasma is not increased;

- the observed increase in the free radical activity of the blood plasma of patient O. is 457%, which is less than 500%;

which indicates the absence of a tumor of a high degree of malignancy.

Preliminary diagnosis: a tumor (glioma) of a low degree of anaplasia.

The patient was operated on 12.12.16 - removal of the tumor.

Histological examination from 12/21/16: histological picture and tumor immunophenotype correspond to fibrillar astrocytoma, ICD-O code 9420/3, grade 2.

Thus, the patient on the 9th day in the postoperative period was diagnosed with low grade glioma.

Example 2

Patient B., 71 years old. Was admitted to the hospital with a diagnosis of volumetric formation of the left temporal lobe.

Complaints: severe headache, nausea, speech impairment.

Anamnesis: has been ill for 2 months, when a headache began to bother, speech disorders joined for about a month. MRI revealed a temporal lobe tumor.

Neurologically: the consciousness is clear, oriented, critical, elements of sensory-amnestic aphasia, anisoreflexion D> S, a positive Barre symptom on the right, a fall in the Romberg position.

According to MRI with contrast enhancement from 12/06/16, a 6X6 cm tumor in the posterior frontal region is spreading in the direction of the Wernicke zone with the cystic component. Upon admission to the hospital before the start of therapeutic measures, an analysis of the biochemical parameters of the blood of patient B. was performed, 71 years old: catalase activity = 5.48 mM / L, acetylcholinesterase = 11.4 mg / ml / hour / 0.1 gNv, myoglobin concentration = 67 ng / ml and free radical activity Imax = 9.01 mV. In this way:

- the observed decrease in erythrocyte catalase activity of patient B. is equal to 64.05%, which is more than 33%;

- the observed decrease in the blood acetylcholinesterase activity of patient B. is 53.20%, which is more than 35%;

- the observed increase in the content of myoglobin in the blood plasma of patient B. is 347%, which is more than 25%;

- the observed increase in the free radical activity of the blood plasma of patient B. is 621%, which is more than 500%;

which indicates the presence of a tumor of a high degree of malignancy.

Preliminary diagnosis: a tumor (glioma) of a high degree of malignancy.

Operation 12/20/16. - removal of the tumor. The postoperative period was uneventful. Conducted anti-inflammatory, analgesic therapy: Amoxiclavi 1.2 g intravenously, Ketonali 2 ml IM.

Histological examination of 12.29.16 histological picture of glioblastoma, ICD-O code 9440/3, grade 4.

Thus, the patient on the 8th day in the postoperative period was diagnosed with glioma of a high degree of malignancy.

Claims (1)

A method for the diagnosis of malignant neoplasms of the brain, including the study of blood plasma by instrumental methods of research and diagnosis, characterized in that they study the free radical activity and content of myoglobin in blood plasma, erythrocyte catalase, whole blood acetylcholinesterase and with an increase in free radical activity of blood plasma by more than 500% of physiological norm 1.25 ± 0.12 mV and an increase in myoglobin content by more than 25% of the physiological norm of blood plasma myoglobin 22.63 ± 6.33 ng / ml, with a simultaneous decrease in the activity of erythrocyte catalase by more than 33% of the physiological norm 15.24 ± 3.21 mM / L and a decrease in the activity of whole blood acetylcholinesterase by more than 35% of the physiological norm 24 , 36 ± 5.72 mg / ml / hour / 0.1 g HB diagnose malignant neoplasm of the brain.

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