Method for evaluating osmotic fragility

FIELD: medicine.

SUBSTANCE: evaluating osmotic fragility requires 10-fold blood dilution (by 0.02 ml), placing the samples in 5 test tubes with distilled water 0.2 ml containing Ca2+ 2.5 mM. They are kept at room temperature for 30, 45, 60, 90 and 120 s, respectively. The haemolysis process is terminated by adding 6% NaCl by 0.2 ml in the test tubes. Further, a microscopy is used to count unhemolysed erythrocytes to derive a water exposure time whereat the erythrocyte count decreases by 50 % from the reference level (T50).

EFFECT: higher accuracy of evaluating the osmotic fragility and more reliable interpretation of research results.

1 dwg, 3 tbl, 1 ex

 

The proposed development relates to the field of medical biology, namely, laboratory research methods in Hematology and physiology.

Determination of osmotic resistance of erythrocytes (DC) is one of the most accessible in the laboratory diagnosis method for estimating physico-chemical properties of erythrocyte membranes. Change the WEM is observed in a number of diseases such as hemolytic anemia, heart failure (decrease WEM), thalassemia, obstructive jaundice, arteriosclerosis (increase WEM) (ADO ROAD, Ishimov L.M. 1980). In physiology the definition of the wholesale electricity market is used to estimate adrenalectomy erythrocytes (Sominskii V.N., Perch C.V., 1981).

Closest to the proposed development is considered microscopic method Yanovsky (Baptist V.E., coast E.A., 1964). The essence of it is that capillary blood is diluted 200 times with four solutions of NaCl - 0,50%; 0,46%, 0.40% and 3%, and then in the counting chamber to determine the number phenolsulfonic of red blood cells in each sample. It is expressed as a percentage of the original number (i.e. a 3% solution of NaCl). The less remains of erythrocytes at 0.50%; 0,46% and 0.40% NaCl solutions, the lower DC.

The disadvantages of this method is the length of the run, the absence of a standard exposure time of red blood cells, the wrong choice of these g is plasmatechnik environments i.e. concentrations of NaCl, the difficulty in differentiation phenolsulfonic erythrocytes from them Strom-shadows, the complexity of the method associated with the counting of red blood cells, the lack of clear criteria for evaluation of the normal and the modified DC. Method has not found wide application in clinical and experimental practice.

There is a combined method of Wekesa (Baptist V.E., coast E.A., 1964). Capillary blood bred hypotonic NaCl solutions(0,42%, 0,46%, 0,52%, 0,62%, 0,70% and 0.82%) and after 6 hours in each sample count of red blood cells in the counting chamber. Measure DC is the concentration of NaCl at which erythrocytes are not already destroyed. The disadvantages of the method are the long duration of the observations, large (6) the number of media needed to determine the wholesale electricity market, the difficulty in differentiation phenolsulfonic erythrocytes from them Strom-shadows, the complexity of the method associated with the counting of red blood cells, the lack of clear criteria for evaluation of the normal and the modified DC. Method due to the complexity and low accuracy has not found application in clinical and experimental practice.

A method for determining the osmotic resistance of red blood cells (4) for changing the diameter of red blood cells placed in a 0.25% solution of NaCl, which is determined at the microscope at 400-fold magnification. Measure DC was the time, for which the diameter of erythrocytes increases in 2 times. The disadvantages of this method include greater complexity, the lack of evidence of the advisability of taking a 0.25% solution of NaCl and the lack of regulatory indicators DC.

There are various ways to determine the osmotic resistance of erythrocytes, based on the assessment of content in the solution of hemoglobin released from erythrocytes in their hemolysis. These methods can be considered as analogues. Among them, a technique based on visual assessment of hemolysis in intensity of colouring of hypotonic NaCl solution; the method is based on measuring the intensity of hemolysis by the degree of light absorption of a certain length; a method based on the direct determination of hemoglobin in a hypotonic environment.

1. Visual, or macroscopic, the method of estimating the DC of Lembeke and Ribera (Baptist V.E., coast E.A., 1964), consisting in the fact that capillary blood (20 μl) is added to a test tube with 10 ml of hypotonic NaCl solution (from 0.56% to 0.28%). The mixture is incubated for 1 hour at room temperature (15-25°C), and then subjected to 3-minute centrifugation at 2000 revolutions per minute, after which mark the maximum and minimum levels of resistance, i.e. the NaCl concentration at which there is a partial and complete hemolysis of erythrocytes. Different is vidrascu this method is a visual assessment of the minimum and maximum stability of erythrocytes during their incubation for several hours in 0.9%, 0,8%, 0,7%, 0,6%, 0,5%, 0,4% and 0.3% NaCl solutions without the use of centrifugation (Baptist V.E., coast E.A., 1964). In General, this method has several disadvantages: the need for a large number of capillary blood, low precision estimates of the minimum and maximum hemolysis, uncertainty reduction degree of toychest solutions, the absence of universally accepted norms DC.

2. Colorimetric method for the estimation of the wholesale electricity market is based on the direct determination of the concentration of hemoglobin, leaving the red blood cells when they hemolysis (Starchenko GO, 1914; Barton JH, 1949). (Baptist V.E., coast E.A., 1964). Originally this method was used geometr Sali, in which the hemoglobin content was estimated by the dilution of hydrochloric acid hematin distilled water to a color standard vials of geometry. The essence of the method lies in the fact that in test tubes with solutions of NaCl(0,9%, 0,8%, ... 0,1%) add the red blood cells, and then determine the method Sali concentration of hemoglobin in each tube, whereby the build curve of hemolysis of erythrocytes with appropriate concentrations of NaCl.

The disadvantage of this method is the low accuracy of determination of hemoglobin by the method of Saly, the complexity and lack of standards.

3. The way Liebelson proposed to determine the WEM by quantifying the degree of hemolysis of eritrotsitov buffered hypotonic NaCl solutions according to the intensity of light absorption wavelength 500-560 nm (green filter) fotoelektrokalorimetry (Menshikov V.V. 1987). In two sterile tubes pre-filled with 2 drops of heparin was added 1.5 ml of venous blood; one sample is used for research, second left on the clock in thermostat at 37°C. In 14 of the centrifuge tubes poured into 5 ml of NaCl solution at a concentration of from 1.0% to 0.10%, and each of them add 0.02 ml of heparinized venous blood; the tubes incubated at room temperature for 30 min, and then centrifuged for 5 min at 2000 rpm, followed by photocolorimetry supernatant (in a cell with a layer thickness of 10 mm) against blank sample, i.e. the supernatant liquid in the test tube containing 1% sodium chloride solution. On the basis of the values of extinction calculate the intensity of hemolysis in each tube by the formula (Ex/E1)×100%, where Ex- the extinction of the supernatant liquid in the test sample, E1- extinction of the supernatant liquid in the test tube with 0.1% solution of sodium chloride (she is 100%). The next day repeat the study with blood, incubated 24 h at 37°C. In healthy individuals, the minimum resistance of erythrocytes is 0.45%-0,50% NaCl solution, a maximum of 0.35%-0,45% NaCl solution. The disadvantages of this method: the need for the use of venous blood in a large volume (3 ml); the complexity of preparation 28 solutions with various con is antracene NaCl, that requires time-consuming and fraught with possible inaccuracies in the hinge-plate; a large number of laboratory ware; a large number of measurements on fotoelektrokalorimetry, which increases the run time of the test, as well as the ability to change the intensity of light absorption depending on the form of hemoglobin in solution (oxyhemoglobin, restored hemoglobin). So far in the literature are not used to such regulations as the percentage of hemolysis based on the calculation of extinction. The literature also discussed the issue of the correct length oxen light when assessing the intensity of light absorption.

4. The method of study of osmotic resistance of erythrocytes (6) assumes that the red blood cells mixed with NaCl(0,9%, 0,7%, 0,5%, 0,3%, 0,1%) directly in the cuvette (10 mm layer) spectrophotometer and immediately at 20°C start recording the optical density at a wavelength of 650 nm to stable values. For 100% hemolysis take maximum readings of the optical density of the suspension of erythrocytes with 0.1% NaCl solution, and for zero hemolysis with 0.9% NaCl solution. Disadvantages of the method are: the availability of special equipment (spectrophotometer Unicam Sp 8000"), as well as the complexity of the method and the absence of indicators of osmotic resistance normal the erythrocytes.

5. The method of determination of osmotic resistance of erythrocytes (7) based on recording the optical density of the solutions of NaCl with 0.1% to 0.9%)containing erythrocytes (5 ml + 0,02 ml of heparinized blood) with the passage of light wavelength 670-750 nm in a cell with a thickness of 10 mm in fotoelektrokalorimetry against blank sample (NaCl solution of appropriate concentration, without red blood cells). The measurement is carried out within a few minutes from the addition of red blood cells, and in contrast to the method Liebelson without centrifugation and 30-minute temperature control at room temperature. The percentage of hemolysis in each tube is calculated by the formula 100 × (Dmax-DX/Dmax-Dmin), where Dmax and Dmin - maximum and minimum optical density, and DF is the optical density of the investigated samples. The transition to wave 670-750 nm increased the accuracy of the method; this method also reduced the complexity, but still the uncertainty in the assessment of regulatory performance DC and the complexity of their interpretation.

6. The method of determination of osmotic resistance of erythrocytes (8), which is a modification of the method Liebelson. In three test tubes (2.5 ml of distilled water, and 0.9% and 0.45% NaCl) add 0.01 ml of capillary blood; the mixture is subjected to 10-minute centrifugation at 2000 rpm, and then evaluate the spectrophotometer PR is the wavelength of 414 nm, the light absorption of the supernatant liquid in a cell with a layer thickness of 10 mm against the cuvette with distilled water. The degree of hemolysis judged by the formula (Tu/EC)×100%, where Tu and EC is the optical density of the supernatant liquid, respectively in vitro with 0.45% NaCl and distilled water (the latter is taken for 100%). Normal DC adult for 0.9% NaCl solution makes 1.61±0,23%to 0.45% NaCl - 25,16±1,83%. Hemolytic States, these figures are respectively 10,13±1,57% and 93,58±is 3.08%. Although the method differs from the method Liebelson less complexity, less cost of laboratory glassware, preparation of solutions. But it has its drawbacks, among which the possibility to change the intensity of light absorption depending on the form of hemoglobin in solution (oxyhemoglobin, restored hemoglobin), as well as insufficient justification of the choice of hypotonic solution (of 0.45% NaCl).

Thus, there are many modifications of the methods of the wholesale electricity market, based on the assessment of content in a hypotonic environment of hemoglobin, including optical density of the supernatant liquid (method Liebelson), numerous modifications which are different wavelengths of light (from 414 nm to 750 nm), different solutions of NaCl and different principles of calculating a measure DC.

The aim of the proposed method is unification evaluation of osmotic resistance of erythrocytes (DC), increase the object is Vesti, the availability and wide use in clinical practice to diagnose pathology of erythrocytes, as well as reducing the complexity, the possibility of estimating the content in the erythrocyte receptors for adrenaline, acetylcholine, histamine and other hormones and biologically active substances according to the nature of changes in the wholesale electricity market in their effects on erythrocytes.

The proposed method enables quantitative determination of the number phenolsulfonic erythrocyte percent of their original content, placed in distilled water at a fixed time (30, 45, 60, 90 and 120 s), with the subsequent calculation time, which in this environment is still 50% of red blood cells (T50).

This goal is achieved by the fact that the red blood cells are placed in distilled water containing 2.5 mm CaCl21 l, on a strictly fixed time (30, 45, 60, 90 and 120 seconds), after which the hemolysis of erythrocytes stops by creating a hypertonic environment by adding distilled water 6% solution of NaCl at a ratio of 1:1, followed by counting the number phenolsulfonic erythrocytes (classical method in the camera Burger with mesh Goryaeva), expressed in percentage to the original level of red blood cells, and determine the duration of the exposure time in water, in which the number of red blood cells is reduced by 50% of the ex the underwater level (T 50).

Implementation of the method lies in the fact that we investigated the generally accepted method is the intake of 0.02 ml of capillary blood (for example, using a micropipette of geometry Sali). This volume of blood is added to a test tube containing 0.2 ml of 0.9% NaCl solution, heparin (1 IU/ml) and CaCl2(2.5 mm); in General, the blood is diluted 10 times. Then 0.02 ml of this dilution is placed in a test tube containing 0.4 ml of 3% NaCl solution; this gives a final dilution of blood in 200 times and allows you to determine the original content of erythrocytes in 1 l of blood (absolute control, taken as 100%). In addition, in 5 test tubes containing 0.2 ml of distilled water with CaCl2(2.5 mm), to be paid to 0.02 ml of 10-fold dilution of blood on a strictly fixed time (respectively at 30, 45, 60, 90 and 120 s) for osmotic hemolysis. Strictly in 30, 45, 60, 90 and 120 in each tube to stop the hemolysis is added 0.2 ml of 6% NaCl (eventually the blood subjected to hemolysis, also diluted 200 times). Then counting the number of red blood cells by the classical method using microscope type LOMO Biolam (R-11) in the counting chamber Alferov-Burker with mesh Goryaeva in 80 small squares. The number phenolsulfonic erythrocytes in each of 5 tubes is expressed in percentage to absolute control. On the basis of these values is based distorting what I (see the drawing), reflecting the rate of hemolysis, which is the duration of the exposure time in water, in which the number of red blood cells is reduced by 50% from baseline (i.e., the rate T50.). The proposed method estimates the WEM can be simplified through the use of only one exposure (e.g., 30 s) without calculation of T50. The method can be used to assess adrenalectomy erythrocytes (or reactivity to other hormones and biologically active substances) on the nature of changes in the wholesale electricity market in distilled water when adding adrenaline in appropriate concentrations, such as 10-9, 10-8, 10-6g/ml.

As an example, here is the data obtained in the study in winter healthy girls who are in folliculinum phase of the cycle. Absolute control were 5.0×1012/l At 30-, 45-, 60-, 90 - and 120-second exposures in distilled water phenolsulfonate left respectively 79%, 73%, 29%, 1% and 0% of the original number; T50- 50 C. In the presence of epinephrine in a concentration of 10-6g/ml, these figures were respectively 86%, 82%, 58%, 1% and 1%, and T50- 61 C. These data allow us to conclude that the adrenaline in the specified concentration increases DC.

Our proposed method estimates the WEM tested in the study of capillary the blood 87 female students of the second year of medical school. It was held during the school year and taking into account the phase of the menstrual cycle, which allowed us to assess the impact on the wholesale electricity market phase of the cycle (table 1) and season (table 2). In these same studies have measured the influence of adrenaline on the wholesale electricity market (table 3). In General, it was found that the source in 80 small squares contained 410,2±7.4 erythrocytes, which corresponds 4,10±0,74×1012erythrocytes in 1 l of blood. After 30-, 45-, 60-, 90 - and 120-second exposures in distilled water remained intact, respectively 76±1,6%, 60±1,7%, 43±1,5%, 20±1,5% and 8.5±1.0% of the initial level; a value of T50amounted to 54.3±1,5 S.

Presents the values proposed to be used as normal values characterizing the WEM. It is shown that DC erythrocytes does not depend on the phase of the menstrual cycle (table 1). At the same time, DC in spring was higher than in winter (table 2). It is established that the adrenaline in concentrations of 10-10g/ml does not affect the wholesale electricity market, at a concentration of 10-8and 10-6g/ml (table 3) improves it.

Table 3
The number of erythrocytes, naturgeschichte hemolysis in distilled water in the presence of epinephrine in a concentration of 10-6g/ml (M±m)
Hypotonicity what I Wednesday The initial number of red blood cells (×1012) in 1 lThe number phenolsulfonic erythrocytes, in % to the original levelT50with
30456090120
Distilled water, DW (n=11)3,86±1175±746±739±813±31±0,343,2±4,1
DV + adrenaline, 10-6g/ml (n=11)3,83±1268±959±10*44±1126±89±453,9±7,9
Note:*the difference with the LW was significantly (p<0.05) as criterion of student

The advantages of our proposed method is the use of microscopic amounts of blood, the possibility of multiple studies (sampling capillary blood), the relative speed of obtaining results, the relative neslon the efficiency of the complete method, high precision evaluation of the WEM, the simplicity and reliability of the interpretation of the results of a study estimating the dynamic destruction of red blood cells and gives the possibility of using this method to assess georectified erythrocytes (adrenergic, holino-, serotonin - and gistaminergicescuu; for these purposes in distilled water is added to ions of CA2+at a concentration of 2.5 mm/l as necessary intermediaries in the action of these substances). The most difficult way is to count the number of red blood cells. Modern microscopes or hemocytometer can greatly facilitate the assessment of the wholesale electricity market for the proposed method. The method can be used in any clinical conditions and in experimental studies related to the assessment of the WEM.

Sources of information

1. Pathological physiology. Edited Addo and Lamisilbuy M.: Medicine, 1980.

2. Sominskii V.N., Perch C.V. Increased osmotic resistance of red blood cells under the influence of propranolol. // Laboratory work. - 1981. No. 9. - S-527.

3. Guide to clinical and laboratory research, based Veeramachaneni edited Eaast. - "Medicine" 1964 - the prototype.

4. Kolomiytsev A.K. Copyright certificate of the Russian Federation No. 2049998 C1, G01N 33/48, published 10.12.1995. Bull. No. 34

5. Laboratory methods in the clinic. Edited Won ikova. M.: Medicine, 1987, p.119

6. Lipina O.V., Shrago M.I., Morozova TV USSR Author's certificate No. 1097949, G01N 33/48, published 15.06.1984. Bull. No. 22.

7. Ryabov, S., Shostka GD, Spiridonov V., DMITRY Bochkov. USSR author's certificate No. 1647403 A1, G01N 33/49 published 07.05.1991. Bull. No. 17.

8. Gorshkov M.A., Miller D.A., Egorov E.N., Fedotov T.A. Copyright certificate of the Russian Federation No. 2328741, G01N 33/49 published 10.07.2008. Bull. No. 19.

The method of evaluation of osmotic resistance of erythrocytes (DC), including the study of capillary blood (0,02 ml), microscopic counting the number phenolsulfonic erythrocytes in hypotonic solution, characterized in that as a hypotonic solution used distilled water containing 2.5 mm CaCl2, as a means of terminating the hemolysis of erythrocytes in a strictly specified time - 6%NaCl solution, and measure the WEM is the percentage phenolsulfonic erythrocytes in 30-, 45-, 60-, 90 - and 120-second exposures (or one of them), as well as the duration (in seconds) exposure of erythrocytes in the water, where their number is reduced by 50% from baseline (T50).



 

Same patents:

FIELD: veterinary science.

SUBSTANCE: in newborn calves with initial dyspepsia symptoms blood samples are taken to identify aggregative activity of platelets using solutions of ADP, collagen, ristomycin and adrenalin as inducers by mixing of 0.1 ml of plasma rich in platelets and standardised by platelets quantity on a slide plate with 0.1 ml of one of aggregating agents and identifying time of platelet aggregates appearance in the form of whitish grain visible with the naked eye. The reference is the average result of identified aggregation of platelets in 10 healthy newborn calves. If platelets aggregation acceleration over 7% compared to the reference is detected in newborn calves with initial dyspepsia symptoms, development of toxic dyspepsia is forecasted. Early forecasting of toxic dyspepsia development in newborn calves has signification practical value, since it makes it possible to more thoroughly treat animals.

EFFECT: reduced loss of calves sick with dyspepsia and improved economic efficiency of stock raising.

1 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, particularly to gastroenterology. Diagnosing active opium addiction in patients with chronic viral hepatitis type C is ensured by evaluating: basal, stimulated level and stimulation index of tumour necrosis factor α (TNF-α); basal and stimulated level of interferon-γ (IFN-γ); stimulation index of interleukin-2 (IL-2); basal level of interleukin-4 (IL-4) If basal TNF-α is 12 pg/ml to 50 pg/ml, stimulated TNF-α is 15 pg/ml to 80 pg/ml, TNF-α stimulation index is 0.65 to 1.78, basal IFN -γ is 20 pg/ml to 22 pg/ml, stimulated IFN -γ is 20 pg/ml to 22 pg/ml, stimulation index of IL-2 is 1 to 1.45, basal IL-4 is 9.1 pg/ml to 11.8 pg/ml, active opium addiction is diagnosed.

EFFECT: technique provides higher accuracy of diagnosing active opium addiction in patients with chronic viral hepatitis type C.

3 ex, 1 tbl

FIELD: medicine.

SUBSTANCE: invention refers to medicine. A matrix mass spectrometry technique is used to detect the presence of a residue in an imprint. A powder containing hydrophobic particles of silicon dioxide, as well as a metal, metal nitride, a metal oxide or carbon is applied on the imprint. An average particle diametre of said metal, metal nitride, metal oxide or carbon introduced in the hydrophobic particles of silicon dioxide is 65 to 90 mcm or 400 to 500 nm preferentially. The investigated imprint is lifted from the application area with using a lifting tape and brought in contact with a sample substrate of a mass spectrometre, then the powder is applied on the imprint. The powder is magnetic or paramagnetic and additionally contains a fluorescent or pigmented dye molecule. The mass spectrometry is conducted with using a matrix specified in a group including MALDI-TOF-MS-MS or SALDI-TOF-MS-MS and their combinations. The residue represents an endogenous residue, particularly endogenous and/or exogenous metabolites and a contact residue. The endogenous metabolite represents squalene, while the exogenous metabolite is nicotine metabolite, e.g. cotinine, and the contact residue is the drug cocaine.

EFFECT: method allows executing the imprint residue analysis on a human imprint directly that eliminates it to be replaced.

21 cl, 14 dwg

FIELD: chemistry.

SUBSTANCE: sodium fluoride is added to an analysed sample in amount of 10% of the mass of the biological object and infused twice in 45 minutes with portions of ethyl acetate, the mass of each of which is twice higher than the mass of the biological material. Separate extractions are combined, filtered through anhydrous sodium sulphate. The solvent from the filtrate is evaporated at temperature 50-60°C. The residue is dissolved in a mixture of hexane-dioxane-propanol-2 solvents. Chromatography is performed in a column with silica gel L 40/100 µ using a hexane-dioxane-propanol-2 mobile phase. The eluate fractions which contain the analysed substance are merged. The eluate is evaporated. The residue is dissolved in a mixture of hexane-dioxane-propanol-2 solvents and the analysed substance is determined via high performance liquid chromatography (HPLC) in a 64x2 mm column filled with Silasorb 600 sorbent using a hexane-dioxane-propanol-2 mobile phase and a UV detector.

EFFECT: invention shortens the duration of detecting tetraethyl thiuram disulphide in blood and increases its sensitivity.

3 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: in patients with progressive myopia if glaucoma suspected, lachrymal fluid is examined for the concentration lactic (LA) and pyruvic (PA) acids for the right and left eyes. Then for each eye, hypoxia coefficients are calculated by formulae: HCOD=LAOS/PAOD, HCOS=LAOS/PAOS, where HCOD is the hypoxia coefficient of the right eye; LAOS is the LA concentration in the LF of the right eye, mmol/l; PAOD is the PA concentration in the LF of the right eye, mcmol/l; HCOS is the hypoxia coefficient of the left eye; LAOS is the LA concentration in the LF of the left eye, mmol/l; PAOS is the PA concentration in the LF of the left eye, mcmol/l. If the hypoxia coefficient HCOD exceeds 1.0, glaucoma on the corresponding eye is diagnosed.

EFFECT: use of the technique enables high reliable diagnosis of glaucoma in patients with progressive myopia.

2 ex

FIELD: medicine.

SUBSTANCE: diagnostic technique for chronic viral hepatitis type C associated with opium addiction consisting in needle biopsy of a liver that is followed by optical microscopy to analyse the prepared biopsy materials for a basal, stimulated level of interleukine-10 (IL-10), and if observing the basal IL-10 within 218.5 pg/ml to 288.5 pg/ml, and the stimulated IL-10 within 468 pg/ml to 588 pg/ml, the presence of fibrosis of the central vein, then chronic viral hepatitis type C associated with opium addiction is diagnosed.

EFFECT: improved diagnostic accuracy.

2 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to diagnosing malignant processes in a human body. A diagnostic technique for a malignant process in a human body consisting in sampling a patient's living tissue, grinding, mixing with physiologic saline to a state of suspension, keeping and agitating under certain conditions, then centrifuging, separating supernatant and detecting cancer-specific markers by an immunochemiluminisent assay.

EFFECT: method exhibits a simple and high degree of malignant process detection in the human body, both at the early, and following stages of disease.

1 tbl

FIELD: medicine.

SUBSTANCE: method of hepatic fibrosis assessment in patients with chronic viral hepatitis type C consisting in evaluating CB56+ phenotype in blood lymphocytes with the value range of blood CD3+/CD56+, CD3+/CD56+/CD4+, CD3+/CD56+/CD8+, CD56+/CD94+, CD56+/NKG2D+, CD56+/CD107a+ cell percentage showing third stage (pre-cirrhotic) or cirrhosis.

EFFECT: method allows higher diagnostic accuracy in fibrous hepatic changes.

7 dwg, 3 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: determining individual genome sensitivity to radon exposure is ensured by genetic blood test to identify predisposing and protective genotypes: marker Arg280His of gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/His; marker Argl94Trp of gene XRCC1 - predisposing genotype Arg/Arg, protective genotype Arg/Trp; marker Asnl48Glu of gene APE1 - predisposing genotype Glu/Glu, protective genotypes Asn/Asn, Asn/Glu; marker A2455G of gene CYP1A1 - predisposing genotype A/G, protective genotypes A/A and G/G; a deletion marker in gene GSTM1 - predisposing genotype o/o, protective +. High individual sensitivity to high radon dose is stated by observing the quantitative prevalence of predisposing genotypes or the equal quantity of predisposing and protective genotypes. High individual resistance to high radon dose is determined by the quantitative prevalence of protective genotypes.

EFFECT: use of the method allows estimating genetically determined predisposition to formation of high level of chromosomal aberrations even before exposing to a radiating factor.

5 tbl, 2 ex

FIELD: medicine.

SUBSTANCE: patient examination involves visual evaluation of a periodontal pocket depth, a tooth mobility degree, gum tissue state with using periodontal indexes. X-ray examination aims at evaluating an alveolar bone resorption degree. It is followed with biochemical blood analysis for the parathormone and calcitonin concentrations. If simultaneously observing the blood calcitonin concentration less than a lower physiological norm, while the parathormone concentration exceeding an upper physiological norm, the presence of chronic aggressive generalised periodontitis is concluded.

EFFECT: use of the technique allows advanced detection of chronic aggressive generalised periodontitis, early identification of said disease among the other inflammatory diseases of periodontium at initial morbidities.

3 dwg, 2 tbl, 2 ex

FIELD: medicine, psychiatry.

SUBSTANCE: one should isolate DNA out of lymphocytes of peripheral venous blood, then due to the method of polymerase chain reaction of DNA synthesis one should amplify the fragments of hSERT locus of serotonin carrier gene and at detecting genotype 12/10 one should predict the risk for the development of hallucino-delirious forms of psychoses of cerebro-atherosclerotic genesis.

EFFECT: more objective prediction of disease development.

3 ex

FIELD: medicine, urology.

SUBSTANCE: one should conduct subcutaneous prevocational tuberculin test and, additionally, both before the test and 48 h later it is necessary to perform the mapping of prostatic vessels and at decreased values of hemodynamics one should diagnose tuberculosis. The information obtained should be documented due to printing dopplerograms.

EFFECT: more reliable and objective information.

1 ex, 1 tbl

FIELD: molecular biology.

SUBSTANCE: the suggested innovation deals with the fact that nucleic acids should be isolated directly out of the sample without pipetting stage but with the help of interconnected reservoirs being prepared beforehand. The above-mentioned vessels should be applied either separately or being interconnected according to standard microtitrating format. The sample should be mixed with a lyzing buffer and nucleic acids are bound with matrix in closed system including, at least, two interconnected reservoirs. Forced movement of sample's mixture and buffer back and forth from one reservoir into another one for several times through narrow passage provides their thorough intermixing. The method provides quick and safe isolation of nucleic acids.

EFFECT: higher efficiency.

44 cl, 4 dwg, 1 ex

FIELD: medicine, phthisiology, microbiology.

SUBSTANCE: diagnostic material is poured preliminary with chlorohexidine bigluconium solution, homogenized, kept at room temperature for 10-12 h and centrifuged. Precipitate is poured with Shkolnikova's liquid medium, incubated at 37oC for 3 days, supernatant part of Shkolnokova's medium is removed, fresh Shkolnikova's medium is added, and precipitate is stirred and inoculated on the dense cellular egg media. Sensitivity of the strain is determined in 3 weeks by the presence of growth in the control tube only. Invention provides enhancing precision and reducing time for assay. Invention can be used in assay for medicinal sensitivity of tuberculosis mycobacterium.

EFFECT: improved assay method.

3 ex

FIELD: medicine, biotechnology, pharmacy.

SUBSTANCE: invention relates to agents used for treatment of pathological states associated with disorder of synthesis of neuromediating substances. Method involves the development of pharmaceutical composition and a method for it preparing. Pharmaceutical composition represents subcellular synaptosomal fractions: synaptic membranes, "light" synaptosomes and "heavy" synaptosomes prepared from gray matter of cerebral hemispheres from experimental animals based on the goal-seeking modification of humoral mediators of nerve endings transformed to synaptosomes in development and regression of malignant processes. The composition provides inhibiting the growth of tumor cells, to elevate span-life of patients with ascite Ehrlich's sarcoma, breast adenocarcinoma Ca-755, Wolker's carcinosarcoma-256.

EFFECT: valuable medicinal and anti-tumor properties of composition.

12 cl, 3 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: method involves carrying out microscopic examination of blood serum samples taken from femoral vein and cubital vein. Femoral vein sample is taken on injured side. The examination is carried out before and after treatment. The blood serum samples are placed on fat-free glass slide in the amount of 0.01-0.02 ml as drops, dried at 18-30°C for 18-24 h. The set of pathological symptoms becoming larger or not changed after the treatment in comparison to sample taken before treatment, and morphological picture of samples under comparison taken from the cubital vein showing no changes or being changed to worse, the treatment is considered to be effective.

EFFECT: enabled medicamentous treatment evaluation in course of treatment to allow the treatment mode to be changed in due time; avoided surgical intervention (amputation); retained active life-style of aged patients.

4 dwg

FIELD: medicine, clinical toxicology.

SUBSTANCE: at patient's hospitalization one should gather the data of clinical and laboratory values: on the type of chemical substance, patient's age, data of clinical survey and laboratory values: body temperature, the presence or absence of dysphonia, oliguria being below 30 ml/h, hemoglobinuria, erythrocytic hemolysis, exotoxic shock, glucose level in blood, fibrinogen and creatinine concentration in blood serum, general bilirubin, prothrombin index (PTI), Ph-plasma, the state of blood clotting system. The state of every sign should be evaluated in points to be then summed up and at exceeding the sum of points being above "+20" one should predict unfavorable result. At the sum of "-13" prediction should be stated upon as favorable and at "-13" up to "+20" - prediction is considered to be doubtful.

EFFECT: higher accuracy of prediction.

2 ex, 3 tbl

FIELD: medicine, juvenile clinical nephrology.

SUBSTANCE: disease duration in case of obstructive pyelonephritis should be detected by two ways: either by detecting the value of NADPH-diaphorase activity, as the marker of nitroxide synthase activity in different renal department and comparing it to established norm, or by detecting clinico-laboratory values, such as: hemoglobin, leukocytes, eosinophils, urea, beta-lipoproteides, lymphocytes, neutrophils, the level of glomerular filtration, that of canalicular reabsorption, urinary specific weight, daily excretion of oxalates, arterial pressure, and estimating their deviation against average statistical values by taking into account a child's age.

EFFECT: higher efficiency of detection.

7 dwg, 1 ex, 6 tbl

FIELD: medicine, urology.

SUBSTANCE: the present innovation deals with differential diagnostics of prostatic cancer and other prostatic diseases at the stage of primary inspection. The method includes the detection of PCA and calculation of probability coefficient for prostatic cancer (PCC) by the following formula: where e - the foundation of natural logarithm (e=2.718…), PCA - the level of total blood PCA in ng/ml, V - patient's age in years. At PCC value being above 0.2 one should diagnose prostatic cancer and to establish final diagnosis one should perform polyfocal prostatic biopsy. The method enables to increase accuracy of diagnostics at decreased number of unjustified prostatic biopsies.

EFFECT: higher efficiency of diagnostics.

2 ex

FIELD: medicine, biology.

SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.

EFFECT: improved an valuable properties of nutrient medium.

3 ex

Up!