Method for evaluating severity of disturbed red blood cell aggregation

FIELD: medicine.

SUBSTANCE: blood plasma is examined in 4 minutes after the beginning of spontaneous red blood cell aggregation for free red blood cell count and cell count in aggregates. A percentage of non-aggregated red blood cells (PNA RBC) by formula PNA RBC=FRBSC×100/(TRBCA+FRBSC) wherein FRBSC is the free red blood cell count, TRBCA are total red blood cells in aggregates. If the PNA RBC is 56 to 30%, I degree of severity is stated, 30% to 4% - II degree of severity, less than 4% - III degree of severity.

EFFECT: use of the invention enables objectifying and increasing precision of evaluation of red blood cell aggregation, evaluating an intensity of patient's microcirculation disorders in a relatively short time, and thereby ensuring well-timed adequate complex of therapeutic measures or corrected therapy.

3 ex

 

The invention relates to medicine, namely to internal medicine, and can be used for rapid assessment of disorders of microcirculation.

In the last decade, both abroad and in Russia, significant progress has been made in the creation of fundamentally new approaches in the study of rheological properties of blood in clinical practice. The need for simple and informative diagnostic tests for detection of intravascular disorders of microcirculation is determined by the needs of practical health care in the problem solving methodology of diagnosis and correction of hemorheological disorders in patients with different pathologies, as well as to predict outcomes and monitoring the effectiveness of therapy.

One of the most important pathological phenomena that occur in the microcirculation system, is an intravascular aggregation of blood cells /mainly erythrocytes/(Firsov N.N., Lassie IV, Priezzhev A.V. Modern problems of whole blood aggregometry. Thrombosis, hemostasis and rheology. 2000, №2 (2), P.9-11; Ants L.V., I.A. Tikhomirov, D.V. Borisov analysis of the impact of plasma and cellular factors on the aggregation of erythrocytes of different age populations. Human physiology, 2002, V.28, No. 4, S-148). In this regard, the current study agregats the organizational capacity of erythrocytes and its correct interpretation.

The basis of the pathogenesis of many diseases and their complications are the processes violations of aggregation of red blood cells, initiating the development of thrombosis in the vessels. The need to study aggregation of erythrocytes is apparent and standardized instruments are missing. In different laboratories, there are various ways of assessing the aggregation of red blood cells, which complicates the interpretation of the resulting data. Frequently used method for quantitative determination of RBC aggregation is a photometric method for detecting decrease in the intensity of light scattered by blood after cessation of stirring the sample in a special cuvettes. Devices using this effect have been developed in our country (Levtov VA, Levkovich YU.N., Ashkenazi IA, I. Potapova About the study of the aggregation properties of blood. - Human physiology, 1978, V. 4, No. 3, s-513.; Lysov VA, Belousov SHE, Parfenov, AS the Method of determining the aggregation of erythrocytes. - Cardiology, 1978, No. 8, p.15-21; Sageline, GI Nazarenko, V. Ssize. Clinical aspects of the microcirculation. L., Medicine, 1985, s-106)and abroad (Schmid-Schnnbein N., Eline £A, Tolger E, Velocity of red cell aggregation (RCA): photometric determination of the half-teme and aggregation constant.-Bibl. Anat, 1975. N 13, p.71-92; Firsov N. N., A. V. Priezzhev, Ryaboshapka O., Sirko J.V. Aggregation properties of erythrocytes of whole blood under shear stress by backscattering nephelometry. Proc.of SPIE, 1993, v. 1884, p.283-290). However, dannysoar requires special equipment and involves complex calculations, so not allows the physician to quickly assess the severity of disturbances of microcirculation of the patient and, accordingly, to make adjustments in treatment. The lack of standardized instruments leads to differences in the geometry of the flow and in the ways of the Desk leaving a blood sample light that leads to the difference of the obtained results.

Widely known direct optical method for the determination of RBC aggregation, recommended for use by who expert Committee for standardization in Hematology (International committee for standardization in hematology (expert panel on blood theology). Guidelines on selection of laboratory tests for monitoring the acute phase response. J. Clin. Pathol. 1988., V.41, P.1203-1212). This method is convenient and simple in execution and meets the following requirements: erythrocyte aggregation is taken into account in the conditions excluding its artificial amplification, the aggregation is taken into account when the standard initial volumetric ratio of the mass of red blood cells and plasma, approaching physiological, for cultivation plasma-suspended red blood cells (and at the same time "fixing" units) use the same suspensorium environment (plasma±aggregation substance), quantitative determination of aggregation in the cell (Goriaev) allows you to simultaneously estimate the size and shape of aggregates visually (Fernandez de CM. et al., 1989).

To improve the measurement accuracy of the detail of measured parameters, evaluation vergennes the violations and the aggregation of erythrocytes have a question about the classification of the aggregation violations by severity. Attempt of classification of hemorheological disorders described in Firsova (Firsov N.N., Korotayev T.V., Vislova M.A. classification of the severity of hemorheological devices. Sat. "Rheological studies in medicine" (v.2), M, 2000, s-141). Taking as a prototype of the proposed Firsova method of separation of hemorheological disorders in degree I, II and III severity, we offer as a defining indicator of the degree of aggregation of violations, take the percentage of non-aggregated RBCs. This indicator is available and easy to identify in any laboratory, quite informative and does not require high material costs.

In the work of N.N. Firsova (2000) describes the principle of separation of severity. The basis of the average value (M), obtained from healthy donors, and standard deviation (σ), which determines the scattering parameter around the mean. Standard deviation is a parameter that reliably separates one average value from the other, a distance doubled its size (2σ). In a normal distribution in the interval M±σ is about 70% of all cases, therefore, the overlap between neighboring distributions is only 15%.

The technical result of the proposed method of assessing the severity of the violation aggregation Erythro itow is that as a defining indicator of the severity of the aggregation disorders, calculate the percentage of non-aggregated RBCs (POE) according to the formula, and when POE from 56 to 30% I set severity, from 30% to 4% - II severity, less than 4% - III severity (see table).

A method of evaluating the severity of the violation aggregation of erythrocytes is that stable blood after 15 minutes of centrifugation at 3000 rpm divided by the plasma and the formed elements. The introduction of 0.2 ml were excavated plasma in plastic tablets, then add 0,02 ml washed in phosphate buffer RBC mass. After 4 minutes after the onset of spontaneous aggregation was carried out by counting the number of free (non-aggregated) of red blood cells using a light microscope in the cell Goryaeva, as well as the number of cells in aggregates. Counting was performed in 2 large (32 small) squares camera Goryaeva. Determined the percentage of non-aggregated RBCs (POE) according to the formula POE=CTU×100/(sea+CTU), where CTU - free erythrocytes, sea - sum of all cells in the aggregates.

Using the inventive method us when hemorheological survey 66 healthy people identified the average value and the standard deviation of the percentage of non-aggregated RBCs, sostaviv what I 69,0±13,0%. Thus, the distribution of the severity of the following: rate of 82%-56%, I degree 56%-30%, II degree 30%-4%, third degree less than 4%. Among surveyed 66 healthy individuals POE is within the normal range were observed in 42 people (64%), I severity - 24 (36%).

Of the 86 patients with essential hypertension without organs targets normal POE was observed in 24 (28%>), I the degree of violation of aggregation of erythrocytes was observed in 49 (57%), II degree-13 (15%). Of the 75 patients with ischemic heart disease and myocardial infarction normal POE was observed in 6 patients (8%), first degree violation of aggregation of erythrocytes was observed in 37 people (49%), II degree-30 people (40%), grade III in 2 (3%)

Clinical example 1

Patient L., 43 years old were treated in the medical ward of the clinic Yugma GOU VPO University with 20.10.2007, 4.11.2007, with a diagnosis of hypertension Art. I, AG 2 tbsp., dyslipidemia, risk 3. Hypertensive history 2 years.

When hemorheological examination level POE was 42,86%, which refers to I the severity of the aggregation disorders. Other rheological parameters are as follows: average unit size - 6,07; the rate of aggregation is 1.91; blood viscosity at the shear rate of 200 s-1- 4.5 MPa·s, at a speed of 50-1-5,5 MPa·s at a shear rate of 10-1

Clinical example 2

Patient B., 60 years were treated at the cardiology Department of the MUSES "the 3rd hospital" with 13.10.2007 on 2.11.2007 years with a diagnosis of ischemic heart disease, acute Q-myocardial infarction anterior-septal area of the left ventricle from 13.10.2007 year. Hypertension Art. III, HSN 2 tbsp., FC 3, diabetes mellitus type 2, moderate, decompensation, risk 4. CHD diagnosed in 2006.

When hemorheological examination level POE amounted comprised 17.54%, II refers to the severity of the aggregation disorders. Other rheological parameters are as follows: average unit size - 7,12, the rate of aggregation - 3,43, blood viscosity at the shear rate of 200 s-1- 5.1 MPa·s, at a speed of 50-1- 6,2 MPa·s at shear rate 10s-1-10,1 MPa·s, the viscosity of the plasma - 1.8 MPa·s; hematocrit - 41,2%.

Clinical example # 3

Patient L., 67 years of age, was treated at the cardiology Department of the MUSES "the 3rd hospital" with 14.10.2006 on 12.11.2006 years with a diagnosis of coronary artery disease, recurrent Q-myocardial infarction anterior-septal area of the left ventricle from 14.10.2006 years, complicated by pulmonary edema. Hypertension Art. III, HSN 2A Art., FC 3, diabetes mellitus type 2, moderate, decompensation, risk 4. CHD diagnosed in 2002.

When hemorheological examination POE was - 3.97%, which refers to III severity aggregation disorders. Other rheological parameters are as follows: average unit size - 11,46, the rate of aggregation - 8,10, blood viscosity at the shear rate of 200 s-1at 5.6 MPa·s, at a speed of 50-1to 6.5 MPa·s at a shear rate of 10-1is 19.2 MPa·s, the viscosity of plasma is 2.7 MPa·s; hematocrit - 49,8%.

Thus, these examples demonstrate the feasibility of using the percentage of non-aggregated RBCs to assess the severity of the violation aggregation of erythrocytes.

A method of evaluating the severity of the violation aggregation of erythrocytes, including the definition of clinical and laboratory parameters, characterized in that in the blood plasma after 4 min after the beginning of the spontaneous aggregation of red blood cells to determine the number of free cells, and the number of cells in aggregates, then calculate the percentage of non-aggregated RBCs (POE) according to the formula POE=CSE·100/(sea+CTU), where CTU - free erythrocytes, sea - sum of all cells in the aggregates, and POE from 56 to 30% - I set the severity, from 30% to 4% - II the severity, less than 4% - III severity.



 

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2 ex, 1 tbl

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