RU2266545C1 - Method for diagnosing atherothrombotic ischemic stroke subtype - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: method involves selecting cell suspension enriched in leukocytes in 50 000 cells per 1 mcl concentration from venous blood of a survived ischemic stroke patient. The suspension is cultured in sterile flasks during 62 h. Supernatant fluid is collected and general cholesterol length is measured. The value being greater than 0.150 mmole/l, atherothrombotic ischemic stroke subtype is to be diagnosed.

EFFECT: high accuracy of diagnosis.

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Description

The invention relates to medicine, namely to neurology, and can be used for retrospective diagnosis of an atherothrombotic subtype of an accomplished stroke, which is necessary for pathogenetically substantiated prophylaxis of repeated cerebrovascular accident.

A known method for the diagnosis of atherothrombotic subtype of stroke based on the totality of clinical data and ultrasound results (duplex scanning) of the carotid, vertebral and intracranial arteries (Stroke. Principles of diagnosis, treatment and prevention / Edited by N.V. Vereshchagin, M.A.Piradova, Z .A.Suslina. - M .: Intermedika. - 2002. - P.113). The main indicator in this case is the presence of an atherosclerotic lesion of the extra- and / or intracranial arteries in the form of a pronounced stenosing, occlusal process, an atherosclerotic plaque with an uneven surface, with an adjacent thrombus on the side corresponding to a focal brain lesion (ibid., P. 63).

The disadvantages of the prototype: duplex scanning requires expensive equipment, which limits its use. In addition, it does not give an accurate idea of the state of intracranial and vertebral arteries, which reduces the accuracy of the method as a whole.

The invention is aimed at solving the problem: improving the availability and accuracy of the method.

The solution to this problem is achieved by examining the patient's venous blood in the early recovery period of ischemic stroke. After culturing the cell suspension enriched in leukocytes for 62 hours at 37 ° C under sterile conditions, the concentration of total cholesterol in the supernatant is determined. When the value of this indicator is more than 0.150 mmol / l, an atherothrombotic subtype of ischemic stroke is diagnosed.

The method is as follows.

A patient taking ischemic stroke takes blood from the ulnar vein in an amount of 10 ml into a sterile tube containing a solution of heparin (500 PIECES). Cell suspension containing leukocytes is isolated by sedimentation in a polyglucin solution. The ratio of whole venous blood and polyglucin solution is 5: 3. The cell suspension is washed twice (once in physiological saline of sodium chloride and once in Eagle’s medium), resuspended in the culture medium, the number of leukocytes and red blood cells in the Goryaev’s chamber is counted, the leukocyte concentration is adjusted to 50,000 in 1 μl (erythrocytes no more than 0.6-0.8 million in 1 μl) and placed in sterile vials of 0.2 ml.

Incubated for 62 hours at 37 ° C. The amount of total cholesterol in the supernatant is determined by the cholesterol oxidase method. To do this, in order to increase the sensitivity of the method when determining low concentrations of total cholesterol, the ratio of the studied biological fluid (serum, supernatant) and the working solution of the reagents, specified in the description of the method by the manufacturer (LACHEMA) 1: 101, is changed to a ratio of 1:10 , and calculate the total cholesterol content as the ratio of the optical density of the experimental sample and the standard sample, multiplied by a factor of 0.517.

Diagnosis of an atherothrombotic subtype of stroke is carried out in the presence of appropriate clinical manifestations and an increase in the level of total cholesterol released by leukocytes above 0.150 mmol / L. With other subtypes of ischemic stroke, a significant increase in this indicator relative to the corresponding age control group does not occur.

Examples of specific performance.

Example 1

Patient Ch., 53 years old, was admitted to the clinic for rehabilitation treatment 11/19/03 with complaints of weakness in the right extremities, which developed upon awakening in the morning of 10/22/03. After 5 days, the strength in the arm and leg began to recover. Upon admission - mild right-sided hemiparesis and hemihypesthesia on the right. I did not measure blood pressure before the disease. In the process of dynamic observation - arterial hypertension 3 tbsp. with changes in the fundus, ECG. When duplex scanning of the main arteries of the head pathology was not detected. On a computer tomogram of the brain - lacunar infarction in the subcortical parts of the left hemisphere.

In the study of total cholesterol released by leukocytes, 12/17/2000 the result was 0.137 mmol / L.

Diagnosis: hypertension 3 tbsp., Lacunar (non-atherothrombotic) stroke in the left hemisphere, early recovery period.

Example 2

Patient D., 50 years old, was admitted to the hospital on 01/13/2004 with complaints of weakness in the right limbs, difficulty speaking. Symptoms developed within a few hours of December 6, 2003. On admission, mild right-sided hemiparesis, hemihypesthesia, motor aphasia. Blood pressure 120/80 mm Hg On a computer tomogram of the brain - on the left at the border of the frontal and temporal lobes, the site of ischemia is determined. Duplex scanning of the main arteries of the head revealed a diffuse atherosclerotic lesion of the common carotid artery on the left, stenosis of the common carotid artery on the left, occlusion of the internal carotid artery on the left.

In the study of total cholesterol released by leukocytes, 02/08/04 the level was 0.258 mmol / L.

Diagnosis: atherosclerosis of the main arteries of the head, occlusion of the left internal carotid artery, atherothrombotic stroke in the basin of the left middle cerebral artery, early recovery period.

Example 3

Patient U., 55 years old, was admitted to the clinic on 6.01.04 with complaints of mild weakness in the right limbs, which developed on 12/11/03 within a few minutes. She reached the degree of plegia, was accompanied by a speech disorder and a decrease in visual acuity in the left eye. He did not suffer from arterial hypertension; at the time of the development of a stroke, his blood pressure was 190/100 mm Hg. Objectively at admission - a light optic pyramidal syndrome. Duplex scanning of brachiocephalic vessels revealed small plaques with signs of instability in the bifurcations of both common carotid arteries without stenosis.

When examining the level of cholesterol released by leukocytes, 02/05/04 the level was 0.200 mmol / L.

Diagnosis: atherosclerosis of brachiocephalic vessels with signs of process instability. Aterothrombotic stroke in the pool of the right internal carotid artery. Early recovery period.

The positive effect of the proposed method consists in the use of an additional new diagnostic criterion for the atherothrombotic subtype of ischemic stroke - the level of total cholesterol released by leukocytes, which increases the availability and accuracy of diagnosis. This indicator has clear quantitative criteria, reflects the participation of certain pathogenetic factors, which has the advantage of determining patient management tactics in the recovery period for the prevention of re-stroke.

Claims (1)

A method for diagnosing an atherothrombotic subtype of ischemic stroke, characterized in that a cell suspension enriched in white blood cells at a concentration of 50,000 in 1 μl is isolated from the venous blood of a patient who has had an ischemic stroke, it is cultured in sterile vials for 62 hours, the supernatant is collected, determined in it, the level of total cholesterol, and with its value exceeding 0.150 mmol / l, is diagnosed with an atherothrombotic subtype of ischemic stroke.