RU2328000C1 - Method of early detection of thromboplastin-formation - Google Patents

Abstract

FIELD: medicine; cardiology.

SUBSTANCE: patients are examined for activity and formation time of thromboplastin during three days running provided these systems are tested within multivariate analysis combined with evaluation of general thromboplastin potential. If value X_BiFTP=0.020 and less thromboplastin-formation is considered to be normal, and if value X_BiFTP within 0.021 to 0.030 start of increased thromboplastin-formation is diagnosed proceeding with evident thromboplastin-formation at value X_BiFTP=0.031 and higher.

EFFECT: application of invention enables to avoid complications for various patients.

3 ex

Description

The invention relates to medicine, namely to cardiology and hematology.

Analogues of the proposed method for early detection of thromboplastinogenesis does not exist.

A known method for assessing the activity and time of formation of thromboplastin (Biggs R., Douglas AS, Macparlane RG The formation of thromboplastin in human blood.// J. Physiol. 1953. - vol. 119, No. 1. - p. 89-104.) . However, never before data was obtained by this method for three consecutive days and was not processed in the subsequent system multivariate analysis with the calculation of the total thromboplastin potential.

The aim of the invention is the early detection of growth of thromboplastin formation in various groups of patients.

The essence of the proposed method is that to detect the growth of thromboplastin formation in patients, the activity and time of thromboplastin formation are evaluated for three consecutive days with data processing by a system multivariate analysis and calculation of the total thromboplastin potential.

The method allows to detect early growth of thromboplastin formation in various categories of patients. This allows you to prescribe the appropriate treatment in a timely manner, supporting thromboplastin formation in patients at a normal level. This will significantly improve the quality of life of patients with a risk of thrombosis, reduce their pathological burden, reduce the number of cases of temporary disability, speed up and improve the quality of inpatient treatment, eliminate disability in some cases, and also extend the life and reduce mortality of patients from heart attack and stroke.

The inventive method is as follows.

The principle of the method. The activity and time of formation of thromboplastin from the component of the plates, deprothrombin plasma and serum of the test blood according to the thrombin time of the test plasma in one-minute intervals. When this mixture is incubated with calcium for 3-6 minutes, highly active thromboplastin is formed (Biggs R., Douglas AS, Macparlane RG The formation of thromboplastin in human blood.// J. Physiol. 1953. - vol. 119, No. 1 .-- p. 89-104.).

To determine endogenous thromboplastin, the following ingredients are required:

1) platelet suspension;

2) deprothrombin plasma;

3) serum;

4) calcium chloride.

Reagents:

1) 0.1 M sodium oxalate solution;

2) 1/40 M calcium chloride solution;

3) 0.15 M suspension of barium sulfate;

4) saline solution (0.9% NaCl).

In a silicone tube that contains 0.6 ml of a 0.1 M sodium oxalate solution, 5.4 ml of blood is drawn from a vein with a dry needle. Carefully mix the contents of the tube. At the same time, 1-2 ml of blood is taken into another tube, from which serum is then obtained.

Preparation of a platelet suspension: oxalate blood in a silicone tube is centrifuged for 5-7 minutes at 1500 rpm. The plasma is aspirated with a silicone pipette into another silicone or plastic tube. Platelet-rich plasma is centrifuged for 30 minutes at 3,000 rpm. Platelets settle to the bottom of the tube. A plasma that contains virtually no platelets is aspirated into a separate tube, and physiological saline is added to the platelets, carefully mixed and centrifuged again for 30 minutes at 3000 rpm. Saline is aspirated and the platelet washing procedure is repeated again. After washing the platelets twice, physiological saline is added to their sediment, equal to 1/3 of the amount of oxalate plasma taken for centrifugation. The contents are well mixed, getting a working suspension of platelets.

Preparation of a solution of deprothrombin plasma: 0.5-1 ml of platelet-free plasma, which was collected after centrifugation for 30 minutes at 3000 rpm, is deprothrombinized with a barium sulfate suspension in a conventional manner. Deprothrombin plasma is diluted with saline in a ratio of 1: 4.

Preparation of serum solution: 1-2 ml of venous blood taken in a dry test tube, put in a water bath at 37 ° C for 2 hours. Serum is separated from the blood clot by centrifugation and diluted with saline in a ratio of 1: 8.

Test procedure: 0.1 ml of platelet-free plasma, which remained after centrifugation for 30 minutes at 3000 rpm, is added to a series of agglutination tubes (7-8) in a water bath at 37 ° C (part of this plasma was taken for deprothrombinization). Separately, 0.3 ml of a platelet suspension, 0.3 ml of a solution of deprothrombinized plasma, 0.3 ml of a solution of serum, 0.3 ml of a 1/40 m solution of calcium chloride are poured into an empty tube, also in a water bath, and the stopwatch is turned on. After every minute, 0.1 ml of this mixture and 0.1 ml of a 1/40 m calcium chloride solution are simultaneously introduced into one of a series of tubes that contains platelet-free plasma and another stopwatch is switched on. Shaking the tube, note the coagulation time of the plasma. The minute study is continued until the thrombin time of the plasma is as short as possible. This prothrombin time characterizes the activity of thromboplastin, and the minute at which the mixture coagulated is the time of thromboplastin formation. Normally, thromboplastin is formed at 2-4 minutes, and its activity is 12-14 seconds.

When registering an acceleration in the formation and / or an increase in the activity of thromboplastin, patients are prescribed the necessary treatment.

For each laboratory, a survey of a group of healthy people is necessary in order to find the average standard values with finding S ² for subsequent calculations.

Thromboplastin formation time and thromboplastin activity were determined daily for 3 days for each patient with the calculation of arithmetic mean values, which are used in subsequent calculations with S ² calculations and data processing using a systematic multivariate analysis.

The method of multivariate system analysis allows you to translate multidimensional quantitative characteristics of the process under study with incomparable absolute values into comparable relative values (Application of morphometry and statistical analysis methods in morphological studies. Compiled by M.V. Uglova, B.A. Uglov, V.V. Arkhipov and al., Kuibyshev, 1982.- 47 s). The calculation was made according to the formulas:

- исследуемый параметр,

- нормативный параметр, X_j - относительная разность.Where

- investigated parameter,

is the normative parameter, X _j is the relative difference.

where Pi is the weight coefficient (coefficient of influence), a is a constant factor (in our studies a = 0.1), σ _j is the standard deviation of the value of X _j in relative units.

S _j ² is the variance of the studied parameter X _i , n _i is the number of observations in determining X _i , S ₀ ² is the variance of the normative parameter X ₀ , n ₀ is the number of observations in determining X ₀ .

where X _{Bi OTP} is a value that integrally characterizes the studied process of thromboplastin formation in a given period (in relative units).

If the total thromboplastin potential (OTP) is 0.020 or lower, thromboplastin formation is normal and there is no risk of thrombosis; when X _{Bi OTP} is within 0.021-0.030, the onset of thromboplastinogenesis begins with a risk of thrombosis; with an increase in X _{Bi OTP} to 0.031 and higher, there is a significant increase in thromboplastin formation, and at the same time, the risk of thrombosis becomes the risk of thrombosis, which requires immediate treatment.

Early detection of thromboplastin formation growth is the basis for the timely appointment of an effective treatment aimed at reducing the risk of thrombosis, and a guarantee of improving the quality of life of patients and its extension.

Example 1. Patient K., 56 years old, with a body weight of 79.0 kg, a body mass index of 29.4 kg / m ² , suffering from metabolic syndrome for 12 years, was examined in a planned manner. For three consecutive days, a patient in the morning on an empty stomach took blood from the ulnar vein with a determination of the formation time of 3.7 min, 3.3 min, 2.3 min and thromboplastin activity of 14.0 s, 12.4 s, 12.0 s processing the data obtained by systemic multivariate analysis. According to the calculation results according to the above formulas, it was found: X _j = 0.033, S _j ² = 0.005, σ _j = 0.0414, Pi = 58.82 (for the formation time) and X _j = -0.015, S _j ² = 0.020, σ _j = 0.0455, Pi = 50.00 (for thromboplastin activity). As standard values, X ₀ = 3.0 ± 0.01, (n = 22), S ₀ ² = 0.055 (for the time of formation) and X ₀ = 13.0 ± 0.12, (n = 22), S ₀ ² = 0.0088 (for thromboplastin activity). According to the calculations, the patient found that the total thromboplastin potential was X _{Bi OTP} = 0.011, which indicated normal thromboplastin formation in the patient's body. The patient was recommended to lead a healthy lifestyle and adhere to an individually selected hypocaloric diet (1946.7 kcal). After this, the patient was examined every month for 6 months. Simultaneously with the inspection was performed for three consecutive days to determine the activity and the time of formation of thromboplastin data processing system multifactorial analysis revealed no knots in all thrombocyte formation during 6 months - X _{Bi OTP} rose above 0.018.

The patient was asked to follow the recommendations given to her in the future.

Example 2. Patient A., 58 years old, with a body weight of 94.0 kg, a body mass index of 32.2 kg / m, suffering from metabolic syndrome for 18 years, was examined in a planned manner. For three consecutive days, the patient took blood from the ulnar vein on an empty stomach in the morning with a determination of the formation time of 3.6 min, 3.4 min, 2.4 min and thromboplastin activity of 13.6 s, 13.3 s, 12.3 s processing the data obtained by systemic multivariate analysis. According to the results of calculations according to the formulas presented found: X _j = 0,043, S _j ² _{= 0,009, σ j = 0,0415,} Pi = 58,14 ( for forming time), and X _j = 0,005, S _j ² = 0,003, σ _j = 0.0418, Pi = 57.14 (for thromboplastin activity). As standard values, X ₀ = 3.0 ± 0.01, (n = 22), S ₀ ² = 0.055 (for the time of formation) and X ₀ = 13.0 ± 0.12, (n = 22), S ₀ ² = 0.0088 (for thromboplastin activity). According to the calculations, the patient found that the total thromboplastin potential was X _{Bi OTP} = 0.025, which indicated an increase in thromboplastin formation in the patient's body. The patient was prescribed an individually selected hypocaloric diet (1620.6 kcal), a system of daily individually selected physical activity, and metformin 500 mg 2 times a day. The patient was examined a month after the start of treatment. Simultaneously with the examination, the activity and time of thromboplastin formation were determined three days in a row with data processing by a system multivariate analysis, which showed the normalization of the total thromboplastin potential X _{Bi OTP} . Observation of the patient for 6 months, following the recommendations given earlier, revealed that thromboplastin formation was within normal limits, X _{Bi OTP} did not exceed 0.017.

The patient was asked to follow the recommendations given to her in the future.

Example 3. Patient C, 59 years old, with a body weight of 81.0 kg, a body mass index of 33.4 kg / m, suffers from metabolic syndrome for 30 years, was examined in a planned manner. For three consecutive days, the patient in the morning on an empty stomach took blood from the ulnar vein with a determination of the formation time of 3.4 min, 3.3 min, 2.9 min and thromboplastin activity of 13.8 s, 13.3 s, 12.2 s processing the data obtained by systemic multivariate analysis. According to the calculation results according to the above formulas found: X _j = 0,067, S _j ² = 0,020, σ _j = 0,0419, Pi = 56,82 (for the time of formation) and X _j = 0,008, S _j ² = 0,005, σ _j = 0.0423, Pi = 55.55 (for thromboplastin activity). As standard values, X ₀ = 3.0 ± 0.01, (n = 22), S ₀ ² = 0.055 (for the time of formation) and X ₀ = 13.0 ± 0.12, (n = 22), S ₀ ² = 0.0088 (for thromboplastin activity). According to the calculations, the patient found that the total thromboplastin potential was X _{Bi OTP} = 0.038, which indicated a significant increase in thromboplastin formation in the patient's body. The patient was immediately prescribed an individually selected hypocaloric diet (1513.9 kcal), a system of daily individually selected physical activities, metformin 500 mg 2 times a day and lisinopril 10 mg 1 time per day. The patient was examined a month after the start of treatment. Simultaneously with the examination, the activity and time of thromboplastin formation were determined three days in a row with data processing by a system multivariate analysis, which showed normalization of the total thromboplastin potential - X _{Bi OTP} . Observation of the patient for 6 months, following the recommendations given earlier, revealed that thromboplastin formation was within normal limits, X _{Bi OTP} did not exceed 0.016.

The patient was asked to follow the recommendations given to her in the future.

Using the proposed method for early detection of thromboplastin formation in cardiology and endocrinology will help to avoid many complications in various categories of patients, reduce the number of cases of temporary disability in them, reduce the duration of hospitalization, reduce disability and mortality.

Claims (1)

A method for early detection of thromboplastin formation growth, characterized in that thromboplastin formation duration and thromboplastin formation activity are determined daily for three consecutive days by processing the data obtained by system multivariate analysis and calculation of the total thromboplastin potential: when X _{BiOTP value} = 0.020 or less, thromboplastin _{formation is} considered normal, at X _B from 0.021 to 0.030 mark the beginning of an increase in thromboplastin formation, which turns into pronounced thromboplastin _generation with X _BiOTP = 0.031 and higher.