Method for estimating clinical effectiveness of antiaggregant therapy in patients with acute coronary syndrome
SUBSTANCE: taken venous blood is separated into two samples. The first sample is stabilised with a solution of sodium citrate, the second one - with ethylene diamine tetraacetate. The first sample of whole blood is added with adenosine diphosphate as an aggregation inducer and tested for a peak amplitude of thrombocyte aggregation and a peak amplitude of adenosine triphosphate release profile by impedance method. The second sample is used to measure a fraction of thrombocytes and a fraction of blood corpuscles. It is followed by calculating a thrombocyte aggregation potential index by formula
EFFECT: improving the objective estimation of the clinical effectiveness of the antiaggregant therapy in the patients with acute coronary syndrome, and providing an opportunity for predicting the clinical course of the disease.
1 tbl, 3 ex
The invention relates to medicine and clinical and laboratory diagnosis. The method is designed to build short-term prediction of the effectiveness of combined antiplatelet therapy in patients with acute coronary syndrome (ACS) and allows to estimate the individual risk of recurrent thrombotic events (myocardial infarction with lifting and without ST-segment elevation, unstable angina) in patients on antiplatelet therapy for acute coronary syndrome.
The number of known ways of assessing the effectiveness of antiplatelet therapy based on the assessment parameters cessation of blood flow under conditions of high shear stress in vitro by formation of a platelet "plug" (PFA-100); the study of the adhesion of platelets under the influence of shear stress (IMPACT cone and plate analyzer); the study of the changes on the surface of platelet associated with its activation (flow cytometry, immunohistochemistry markers, such as P-selectin); the study of the reaction of the release of platelets (determination of thromboxane B2 in the serum, the products of its degradation in the urine, indirect determination of the induced arachidonic acid formation of thromboxane A2) [Hezard N, Tessier-Marteau A, Macchi L., Cardiovasc. Hematol. Disord. Drug Targets, 2010; 10 (3): 224-33; Gorlinger K, Jambor C, Dirkmann D, Dusse F, Hanke A, Adamzik M, Hartmann M, Philipp S, Weber AA, Rahe-Meyer N.Herz. 2008 Jun; 33 (4): 297-305.].
One is about describes how difficult and time consuming to use, have a high variability of the estimated parameters, low sensitivity, require special preparation of the sample of blood and experienced operator and expensive to use.
There is a method of predicting the course of acute myocardial infarction (see RF patent №2121691 on CL IPC G01N 33/86, publ. 10.11.1998), including the study of biochemical parameters of blood of the patient in the dynamics and subsequent prediction of unfavorable course of the disease. As the studied parameters using prothrombin index and the unfavorable course of the disease predict when the index decreases from the third to the fourth day from the onset of the disease.
The disadvantage of this method is the low sensitivity and low specificity for assessing the risk of recurrence of thrombotic events. This method is based on the study of prothrombin index, the value of which is determined by the functional activity and the number of coagulation factors II, VII, V and X and the sensitivity of the used device. That is, on the one hand, assessment is used exclusively coagulation component of the blood coagulation system and is not considered platelet mechanism of hemostasis, the impact on which is the essence of antiplatelet therapy aimed at preventing the re-development of the Tr is emboliceskih events. On the other hand, the specified parameter has a high variability and low reproducibility in different laboratories.
There is also known a method of estimating the risk of recurrence of thrombotic events in patients with acute coronary syndrome (see RF patent №2419800 on CL IPC G01N 33/86, publ. 27.05.2011), which consists in determining the level of hematocrit, number of erythrocytes, platelets, determining the time adenosintriphosphate-induced-ADP-induced sportivnie blood, the evaluation values in points.
However, the method does not take into account a number of features of platelets, in particular the characteristic reactions of the release of platelets, their volume fraction in whole blood. The described method is an integral assessment of the hemostatic system and can not adequately assess the risk of recurrent thrombotic events in patients with thrombocytopathy, including drug.
Also known "Method for diagnosis of userinitiated in patients with ischemic heart disease" (see RF patent for the invention №2348041 on CL IPC G01N 33/68, publ. 27.02.2009), for which prior treatment with aspirin platelets peripheral blood of patients determine the activity of the reactions of NAD - and NADP-dependent dehydrogenases. Then calculate the ratio cofactoring exchange platelets (CCOT) formulathat=RBI/IVN, where RBI is the ratio of the activity levels of NADP and NADPH-dependent dehydrogenase reactions, and UN - level activity of NADH-dependent dehydrogenase reaction to the level of activity of NAD-dependent lactate dehydrogenase reaction. And when the value CCOT above 0.3 diagnose aspirinarisoprodol, and a value of KCAT equal to or less than 0.3, - sensitivity to aspirin. The method allows to diagnose aspirinarisoprodol patients before treatment with aspirin and provides a high accuracy of the prediction.
However, the implementation of this method requires considerable cost and extremely time consuming.
Closest to the claimed is a method of evaluating the effectiveness of antiplatelet therapy in the treatment of cardiovascular diseases (see RF patent №2379684 on CL IPC G01N 33/48, publ. 10.06.2009), which consists in determining the aggregation activity of platelets in whole blood impedance method, the determination of the amplitudes of the aggregation process before and after incubation in vitro with acetylsalicylic acid.
However, this method of evaluating the effectiveness antiagregantnoy therapy is not objective, because assessment of disaggregating action of acetylsalicylic acid after incubation in vitro, which does not take into account the impact of its deacetylase in the gastro-kiseon the m path.
On the other hand, the proposed method is based on the analysis of the integral parameter aggregation capacity of platelets and does not take into account the contribution of the reaction releases (status granules of platelets and cell membranes), as well as aggregate-the suspension state of the blood, which ultimately can lead to incomplete accounting of the parameters, leading to the risk of re-occurrence of thrombotic events in patients on antiaggregatory therapy.
The task of the invention is to increase the objectivity of the assessment of the effectiveness of antiplatelet therapy in patients with acute coronary syndrome due to the possibility of a comprehensive record of the aggregate-suspension condition of blood and aggregation capacity of platelets, including the reaction of the release of ATP with the technical simplicity and reducing the cost of the complete method.
To solve the problem in the method of evaluating the clinical efficacy of antiplatelet therapy in patients with acute coronary syndrome, which consists in sampling venous blood, stabilize it, the definition of the aggregation activity of platelets impedance method, the determination of the amplitudes of platelet aggregation according to the invention venous blood is separated into two samples of whole blood, one of which stabilize Rast is the PR of sodium citrate, and the second is a ethylenediaminetetraacetate, in the first sample of whole blood contribute as an inducer of platelet aggregation adenosine diphosphate, determine the maximum amplitude of platelet aggregation and the maximum amplitude of the curve of the release of ATP in the second sample to determine the proportion of the volume occupied by platelets and the proportion of blood volume that is attributable to the formed elements, determine the index of platelet aggregation capacity of blood and when the index value of less than 0.5% judge of the low efficiency of antiplatelet therapy, and when the index value of 1.5-2.5% - of its high efficiency, the index of platelet aggregation capacity of whole blood is determined by the formula
where Lmax- the maximum amplitude of the curve of the release of ATP
Ωmax- the maximum amplitude of platelet aggregation
PCT is the percentage of the volume of whole blood occupied by platelets,
STC - the proportion of blood volume that is attributable to the formed elements.
In well-known authors of the sources of patent and scientific and technical information, not the written method of assessment of the effectiveness of antiplatelet therapy, to objectively and with a high degree of probability to assess the degree of risk of recurrence of thrombotic events in patients with acute coronary syndrome due to the possibility of a comprehensive evaluation of the parameters of the aggregate-suspension condition of blood and aggregation capacity of platelets, including the reaction of the release of adenosine triphosphate.
Known impedance and Lumi-method aggregatometry whole blood (see, for example, W.Heptner, et. al. Thromb. Haemostas. 58, 1676 (1987); P.Sathiropas, et. al. Thromb. Res. 51, 55 (1988)), which allows us to estimate the aggregation activity of platelets in the sample of diluted whole blood in response to the introduction of aggregation inducers, including quantitative parameters of the reaction release Danilovich nucleotides (ATP, nmol). However, as defined by this method the parameters are highly variable, determined by the characteristics of the sample is blood, its type (venous or arterial), patient age, presence of a particular type of thrombophilia and/or thrombocytopathy, background and/or comorbidity, range of used medical therapies, including antithrombotic, concentration, time and method of making aggregation inductor, etc.
There is no standardized parameters estimates, almost not studied for their predictive power.
In the inventive ways the e by studying artificially simulated situation, the formation of a blood clot in the coronary arteries in vitro, the authors were able to find a parameter to evaluate the risk of recurrence of thrombotic events (index platelet aggregation capacity of whole blood), which, according to the Applicant, up to the present time was not known.
The index of platelet aggregation capacity of whole blood followed by its ranking for prognosis of the effectiveness of antiplatelet therapy was not used, because they were not known and were not identified (to the present invention), the interval of values that are characteristic of patients with acute coronary syndrome indicators, reflecting the ratio of aggregate-suspension state of the blood and the reaction of the release of platelet underlying conclusions on the assessment of the risk of recurrence of thrombotic events in this invention.
In addition, using a sample of whole blood, diphosphate as an inducer of platelet aggregation, no stage preanalytical sample preparation allows you to optimize the costs of implementing the method, which generally increases its adaptability and allows to evaluate the efficacy of antiplatelet therapy for more patients.
Claimed by the authors the way technically and technologically with the first manipulation until practically the last operation - processing of the data, since all manipulations: blood sampling, selection and preparation of samples, the effect of reagents on the sample, the incubation time, the procedure is OK action in the claimed method - signs technically actions.
This allows us to conclude about the presence in the invention of the criterion of "inventive step".
The method is as follows.
Taking venous blood, divide it into two samples, one of which stabilize 3,8% solution translesanas of citrate in the standard ratio of 9:1 and immediately assess the aggregation activity of platelets in diluted whole blood by the method of impedancometry with parallel determination of the reaction the release of ATP by the method of lomearogetyj as follows.
1. Include warm up aggregometer to the state of maintaining a stable temperature in the cells 37º in accordance with the instruction on its application.
2. Prepare and warm up the used reagents to 37º in the appropriate cells of aggregometry.
3. When using multi-channel aggregometer may conduct research in duplicate to increase the accuracy of the study.
4. Make each measuring cell of 0.45 ml of saline solution and a magnetic stirrer, and heated for 5 minutes.
5. Contribute to the cell of 0.45 ml of whole blood of the patient, followed by heating for 5 minutes, after which the bolus is made 0.1 ml of phosphor (CHRONOLOG LUME).
6. After sterowania device for STD is bound to the ATP solution and the baseline impedance, in accordance with the instructions for use of aggregometry, bolus way to make a working solution of ATP to a final concentration of 10 μl.
7. Write agregatogramme curve and the release of ATP for 5 minutes (maximum, depending on the time of the release of the designated parameters on the plateau).
8. Determine the maximum amplitude of agregatogramme and curve reactions release (using standard software).
The second part of the sample of whole blood stabilized by ethyleneterephthalate (potassium EDTA) in a standard ratio of 1.2 to 2 mg per 1 ml of whole blood and used for hematological studies using Hematology analyzers - counter flow type (in our case ABXMicros). Determine trombosit (PCT) - the proportion of the volume of whole blood occupied by platelets, and hematocrit (HCT) is the proportion of blood volume that is attributable to the formed elements.
The use of stabilizers whole blood in this version of the principle. Sodium citrate is the most gentle stabilizer in relation to cell membranes. It binds calcium ions in the blood plasma, thereby breaking the cascade of its collapse, which largely determine the kinetics and severity of thrombosis and minimally complexiry with coagulation proteins, so it is the stabilizer of choice for koagulologicheskih studies of blood.
The use of EDTA is fundamentally important to cytomorphometry and hematological analyses. For use of blood, stable EDTA, calibrated almost all used in clinical laboratories Hematology counters.
Evaluation of the effectiveness and prognosis of antiplatelet therapy is performed according to the index of platelet aggregation capacity of whole blood using the following parameters: Ωmax- the maximum amplitude of platelet aggregation, determined by the method of impedancometry, Lmax- the maximum amplitude of the curve of the release of ATP, indicators PCT-thrombocyte and NTS hematocrit (in shares).
The calculation of the index of platelet aggregation capacity of whole blood is done according to the following formula:
This formula was derived from analysis of the results of the study of the dynamics of the aggregation activity of platelets by the method of impedance and lumiere.html in patients with ACS (n=174)found on the standard antiplatelet therapy. The observation period was 12 months. On the basis of the forces in ormation and correlation of all evaluated parameters agregators and clinical outcomes at the end of the observation period, as well as pathophysiological entity clotting of blood was withdrawn index of platelet aggregation capacity of whole blood, which represents the ratio of the total number released during the initial stages of aggregation of ATP to the "prevalence" of platelet aggregation in notional volume of whole blood, expressed as a percentage.
Then is the ranking and rating specified index according to the following diagram for clarity in the table.
|The values of I||The risk of recurrent thrombotic events against the backdrop of ongoing antiplatelet therapy||The need for additional diagnostic action|
|0,00-0,50||High||Verification etiology of thrombophilia|
|1.5 to 2.5||Low||No|
|>2,5||Undefined||Evaluation of the aggregation capacity of platelets with other inducers is gregali|
The risk of recurrent thrombotic events against the backdrop of ongoing antiplatelet therapy was evaluated as a quantitative characteristic of the "risk" of re-occurrence of thrombotic events (this is the ratio of the number of adverse effects (the sum of fatal cases of recurrent non-fatal thrombotic events (n) to their possible number for a certain period (N) R=N(t)/Q(f), where N(t) - a quantitative measure of the frequency of undesirable events in a unit of time t; Q(f) is the number of risk objects (patients included in the study), subject to a specific risk factor (f).
In the ICU, Saratov research Institute of cardiology examined in accordance with the above-described method, the group of clinically healthy individuals and patients with acute coronary syndrome, including unstable angina, myocardial infarction, with the rise and without ST segment elevation. In patients with ACS test results were compared with the clinical outcome of the disease, death, complications, recurrence of the disease. The maximum amplitude of impedancometry for clinically healthy patients ranged from 6 to 18 Ohms at the maximum amount of released ATP from 0.38 to 1.1 nmol that corresponds to the declared reference intervals when used as induct the RA aggregation ADP at a final concentration of 10 Microm. The maximum extent of ADP-induced platelet aggregation ranged from 0 to 21 Ohms in patients with NS and from 0 to 17 Ohms at mi patients, all patients were on standard antiplatelet therapy at the maximum amount of released ATP from 0.0 to 1.71 nmol. Differences in these parameters among patients with and without repeated thrombotic events on the achieved degree of statistical significance, due to the high degree of variance of the data and non-equilibrium proportion of patients in the studied groups. Proposed by the authors, an index of platelet aggregation capacity of whole blood was found statistically significant differences in the group of patients with recurrent thrombotic events and unaccented (ManWhitney p level = 0.041), and revealed a negative moderate correlation relationship between the rank of the outcome and used an index of platelet aggregation capacity of whole blood (Kendall τ 0.69, plevel 0.002), while the variability of the index within each group does not exceed 17.2 per cent. Application of the proposed method of predicting the effectiveness of antiplatelet therapy results (based on the studied sample of ACS patients) to the correct classification of patients in 83.5% of cases.
Clinical example 1. Patient S., aged 65, was admitted to the hospital with a diagnosis of CHD, Q-myocardial infarction posterior wall of the left W ludacka, arterial hypertension stage III, the complication: acute heart failure 1 Killip class. Received standard therapy according to the recommendations of the Committee of experts of the all-Russian scientific society of cardiologists dual antiplatelet therapy, inhibitors of Ha factor, statins, ACE inhibitors, beta-blockers. According to dopplering identified areas of hypokinesis, preserved ejection fraction (61%), signs of congestion at rest no identified diastolic dysfunction. Biochemical indicators of blood count is within normal limits. Ωmax- 7 Ohm, Lmax- 0.01 nmol, PCT - 0,180 and STC - 0,456. The index of platelet aggregation capacity of whole blood of the patient was 0,056 (high risk). According to coronary angiography revealed occlusion of the right coronary artery, hemodynamically insignificant stenosis of the anterior descending artery. Conducted stenting of the right coronary artery with drug-eluting stent with good angiographic results - complete restoration of blood flow in the infarct-related artery. After 3 days after stenting amid continued in accordance with the recommendations of antithrombotic treatment suddenly deteriorated state - developed re-anginal attack, against which it was recorded elevation of ST segment in the front-the side from which edenia. The patient died after 1.5 hours from the onset of pain in the clinical picture of cardiogenic shock.
Clinical example 2. Patient A., aged 63. Was admitted to the hospital with Q-myocardial infarction of the anterior wall of the left ventricle. History of arterial hypertension, a history of myocardial infarction, angina FC III, chronic heart failure 3 tbsp., FC III, diabetes mellitus type II. The disease was complicated by the development of acute heart failure III Killip class, pneumonia, left hydrothorax. Received standard therapy according to the recommendations of the Committee of experts of the all-Russian scientific society of cardiologists dual antiplatelet therapy, inhibitors of Ha factor, statins, ACE inhibitors, beta-blockers, diuretics inside and intravenous inotropic drugs, insulin. If dopplerechocardiography identified areas of Hypo - or akinesia, aneurysm of the apex of the left ventricle, the signs of decompensation on both circles, the ejection fraction of the left ventricle 40%. General analysis of blood without pathology. Upon receipt of biochemical signs of decompensated diabetes. Coronary angiography was not performed. Biochemical indicators of blood count is within normal limits. Ωmax- 5 Ω, Lmax- 0.16 nmol, PCT - 0,201 and STC - 0,455. The index of platelet AGR is haciendolo capacity of whole blood of the patient was 1,413 (medium risk).
On a background of treatment decreased manifestations of chronic heart failure - increased tolerance to stress (at discharge level II FC), disappeared edema of the lower limbs, signs of stagnation in a small circle and hydrothorax (according to repeated x-ray examination), there were no strokes, according to the biochemical analysis of the achieved compensation of metabolic processes. The patient was discharged under the supervision of a cardiologist at the place of residence, within 11 months - no re-thrombotic events.
Clinical example 3. Patient C., 31 years. Was admitted to the hospital with Q myocardial infarction of the anterior wall of the left ventricle, the complication: acute heart failure 1 Killip class. Previously, diseases of the cardiovascular system was not. Received standard therapy according to the recommendations of the Committee of experts of the all-Russian scientific society of cardiology - thrombolytic therapy, dual antiplatelet therapy, inhibitors of Ha factor, statins, ACE inhibitors, beta-blockers. According to dopplering identified areas of Hypo - and akinesis, the ejection fraction of the left ventricle 48%, signs of congestion at rest no. Biochemical indicators of blood count is within normal limits. Ωmax- 14 Ohm, Lmaxis 0.65 nmol, PCT - 0,175 and the STC - 0,371. The index of the platelet and relacionado capacity of whole blood of the patient was 2,19 (low risk). According to coronary angiography revealed occlusion of the anterior descending coronary artery in the proximal segment. Conducted stenting anterior descending coronary artery with drug-eluting stent with good angiographic results - complete restoration of blood flow in the infarct-related artery. The course of the disease without complications. Written under the supervision of a cardiologist at the place of residence.
The above testifies to the adequacy of sufficient sensitivity and usefulness of the proposed method for predicting the clinical efficacy of antiplatelet therapy in patients with ACS.
Medico-social effect of the proposed method is pathogenetically justified sensitive assessment of the risk of recurrence of thrombotic events, i.e. clinical efficacy of antiplatelet therapy in patients with ACS.
First proposed based on the comprehensive assessment of options aggregate-suspension condition of blood and aggregation capacity of platelets, including reactions release, a method for predicting the clinical efficacy of antiplatelet therapy in patients with ACS. In addition, the novelty of the claimed method is the use of specific these four indicators simultaneously in the same method for predicting clinical effectively the ti antiplatelet therapy in patients with ACS.
The economic effect of the proposed method in comparison with currently used is on the one hand to reduce the costs of the research. For example, the average price of the cartridge PFA COLLAGEN/ADP CARTRIG for ways analogous to 20 measurements (two patients) is 10817,3 rubles, i.e. the price of the cartridge for the study per patient is about 1081,73 rubles. While the cost of reagents for the study 1 the patient is about 270,75 rubles, which is much cheaper at the expense of consumables. When compared with aggregatometry whole blood impedance and luminette advantage lies in obtaining additional information about the status of ACS patients with similar personnel, time and resource natrataste.
The method allows prediction of the disease in the first hours of receipt of the patient in any medical institution, which has a laboratory equipped with aggregometer (impedance and lunagraphica) and hematological counter and doctor-assistant, which can handle data of General blood analysis and research aggregation activity of platelets.
The developed technology by the present method extends the Arsenal of tools to objectively monitor the condition of patients with ACS.
Method of assessment of clinical effectiveness of Antia ehentai therapy in patients with acute coronary syndrome,
consisting in sampling venous blood, stabilize it, the definition of the aggregation activity of platelets impedance method, the determination of the amplitudes of platelet aggregation, characterized in that the venous blood is separated into two samples of whole blood, one of which is stabilized solution of sodium citrate, and the second - ethylenediaminetetraacetate, in the first sample of whole blood contribute as an inducer of platelet aggregation adenosine diphosphate, determine the maximum amplitude of platelet aggregation and the maximum amplitude of the curve of the release of ATP in the second sample to determine the proportion of the volume occupied by platelets and the proportion of blood volume that is attributable to the formed elements, determine the index of platelet aggregation capacity of the blood and when the value of the index is less than 0.5% judge of the low efficiency of antiplatelet therapy, and when the index value of 1.5-2.5% - of its high efficiency, the index of platelet aggregation capacity of whole blood is determined by the formula:
where Lmax- the maximum amplitude of the curve you who are aware of ATP,
Ωmax- the maximum amplitude of platelet aggregation
PCT is the percentage of the volume of whole blood occupied by platelets,
STC - the proportion of blood volume that is attributable to the formed elements.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine. A composition for stimulating the skin stem cell production containing interleukin-1 alpha and a dermatologically acceptable diluent or carrier.
EFFECT: invention provides improving the stem cell stimulation.
SUBSTANCE: method involves dissolving 855 mg of a crystalline hydrate of copper chloride (CuCl2·2H2O) in 100 ml of distilled water (concentration of Cu2+ ions in the prepared solution is 50 mmol/l) and adding 1 ml of the prepared solution to 100 ml of a standard reagent used in glucose oxidase test. The ascorbic acid oxidant used is copper chloride solution in end concentration in the glucose oxidase reagent of 500 mcmol/l.
EFFECT: method enables correct determination of glucose content.
1 tbl, 1 ex
SUBSTANCE: workers' blood serum is analysed for interleukin 4, protein S-100β, protein S-100 autoantibodies, voltage-dependent Ca-channel autoantibodies, glutamate receptor autoantibodies, γ-aminobutyrate receptor autoantibodies, dopamine receptor autoantibodies; diagnostic coefficients F1 and F2 are calculated; if the value F1 is less than F2, the early changes of the nervous system are diagnosed for the chronic exposure to vinyl chloride; F1 more or equal to F2 enables stating the absence of any signs of the chronic exposure to vinyl chloride. The developed method may be used in the periodic medical screenings, medical examinations of workers to diagnose some occupational diseases.
EFFECT: use of the invention improves higher accuracy of identifying the various signs of the chronic exposure to vinyl chloride through the use of a complex of the immunological structures of nerve tissue in the chronic exposure to vinyl chloride.
1 tbl, 2 ex
SUBSTANCE: invention may be used to predict a developing myocardial dysfunction in the children with acute lymphoblastic leukemia (ALL) at different stages of polychemotherapy (PCT). The method involves the blood examination for the iron metabolism parameters, namely before the beginning of polychemotherapy (1) and after the induction of remission (2), blood serum ferritin, hepcidin and iron are evaluated in the patients; the derived values are inserted into the equations to calculate varying ECG, IMS, B(E-Ea) NT-pro-BNP after the completion of the intensive PCT course (3) and the total coefficient K is calculated by formula K=ECG3* IMS3* B(E-Ea)3* NT-pro-BNP3, wherein a probability of the myocardial dysfunction is stated by the total coefficient, namely: the coefficient K> 0.24 ensures predicting the developing cardiac complications, while K <0.24 show a lower risk of the cardiac complications.
EFFECT: possibility to detect a risk of the developing myocardial dysfunction accompanying the early PCT by the biochemical parameters, namely in terms of iron metabolism.
1 tbl, 1 ex
SUBSTANCE: method consists in determining a characteristic profile of a test sample of a human biological fluid. It is concentrated off-line. Biologically active substances are separated using complexing additives, and a 'reference' is determined. Steroid hormones are taken as the analysed biologically active substances. The hormones are separated by performed by reversed-phase HELC in gradient elution using a diode array detector. The steroid profiles are used to form a matrix of the analytical signal intensities and the retention factors of each steroid. Each sample is graphically imaged by method of principal components, and the graphical images are used to form 'reference' and deviation clusters. The 'reference' and deviation clusters are corrected by soft independent modelling of class analogy taken as a reference. The pathologies are diagnosed by an ability of the patient's image to come with a 'reference' or a deviation.
EFFECT: reliable diagnosis of the pathologies associated with adrenal cortical diseases.
6 dwg, 2 ex
SUBSTANCE: what is presented is a method for prediction the efficacy of the anti-TNF therapy in the patients with rheumatoid arthritis on the basis of genetic typing the polymorphisms of TNF-alpha proinflammatory cytokine. The allelic polymorphism of the TNF-alpha gene promoter is studied in position 857. If the heterozygous state (genotype - 857ST) or the homozygous T allele carriers (genotype - 857TT) is identified, a high probability of the successful infliximab therapy is predicted. If identifying the homozygous allele C carrier in position - 857 of the TNF-alpha gene promoter (genotype - 857SS), a high probability of the failed infliximab therapy is predicted.
EFFECT: invention enables the rapid and effective prediction of the clinical outcome of the anti-TNF therapy in the patients with rheumatoid arthritis by one polymorph position.
2 tbl, 1 ex
SUBSTANCE: menopausal women with an endometrial hyperplastic type having complaints about spotting undergo biopsy of the lining of the uterus to determine endometrial progesterone and testosterone, if observing progesterone falling within the range of 2.0 to 7.0 ng/g of tissue and testosterone falling within the range of 4.0 to 8.8 ng/g of tissue, developing endometrial cancer is predicted, while progesterone within 24.0 to 29.6 ng/g of tissue and testosterone within 16.8 to 22.4 ng/g of tissue enable predicting developing uterine fibroid. The technical and economic effectiveness of the method consists in the fact that the detected levels of progesterone and testosterone in the intact endometrial tissue in the menopausal patients with a hyperplastic type are high-information laboratory indicators of the presence of either malignant, or benign uterine pathology, which can be used to form the groups of patients with the high risk of malignancy in the body of the uterus.
EFFECT: method is available, quick to implement.
1 tbl, 2 ex
SUBSTANCE: laboratory diagnostic technique for the small-dose poisoning with organophosphorous agents consists in assessing catalytic activity of blood plasma aryl esterase heated to 55°C for 5 min with indophenyl acetate used as a substrate. Catalytic activity of the enzyme and its increase after heating are assessed once after a contact with a organophosphorus agent; the effect of increasing catalytic activity of blood plasma aryl esterase after heating is expressed in %, taking catalyst activity of blood plasma aryl esterase before heating as 100%. In case activity of plasma aryl esterase after heating is increased by more than 20%, the small-dose poisoning with organophosphorus agents is diagnosed.
EFFECT: use of the declared technique enables stating effectively the fact of the small-dose poisoning with OFAs in the absence of any clinical signs of poisoning.
1 tbl, 2 ex
SUBSTANCE: method for prediction the recurrence-free survival period in cervical cancer (CC) is implemented by blood plasma catalase activity test, and the local process of cervical cancer (FIGO stage Ib-IIa) with activity 0.041 to 0.113 mmol/min/l in the patients enables predicting 50% probability of the 18-month period of the recurrence-free survival period, while blood plasma catalase activity 0.008 to 0.035 mmol/min/l shows 80% probability of the recurrence-free survival period.
EFFECT: prediction of the recurrence-free survival period in local cervical cancer.
SUBSTANCE: patient with advanced cervical cancer (FIGO stage IIb-IV) is examined for tumor tissue glutathione reductase activity. When tumour tissue glutathione reductase activity is 10.9 to 14.9 mmol/min/mg of protein, the patient is expected to have 18% probability of the 18-month recurrence-free survival period, while tumour tissue glutathione reductase activity being 15.2 to 18.5 mmol/min/mg of protein shows 60% probability of the 18-month recurrence-free survival period.
EFFECT: use of the declared method enables predicting the recurrence-free survival period in local cervical cancer.
FIELD: medicine, hepatology.
SUBSTANCE: one should detect the level of hepato-specific enzymes (HSE) in blood plasma, such as: urokinase (UK), histidase (HIS), fructose-1-phosphataldolase (F-1-P), serine dehydratase (L-SD), threonine dehydratase (L-TD) and products of lipid peroxidation (LP), such as: dienic conjugates (DC), malonic dialdehyde (MDA). Moreover, one should detect the state of inspecific immunity parameters, such as: immunoregulatory index (IRI) as the ratio of T-helpers and T-suppressors, circulating immune complexes (CIC). Additionally, one should evaluate the state of regional circulation by applying rheohepatography (RHG), the system of microhemocirculation with the help of conjunctival biomicroscopy (CB) to detect intravascular index (II). In case of increased UK, HIS levels up to 0.5 mcM/ml/h, F-1-P, L-SD, L-Td, LP products, CIC by 1.5 times, higher IRI up to 2 at the norm being 1.0-1.5, altered values of regional circulation, increased II up to 2 points at the norm being 1 point, not more one should diagnose light degree of process flow. At increased level of UK, HIS up to 0.75 mcM/ml/h, F-1-P, L-SD, L-TD, LP products, CIC by 1.5-2 times, increased IRI up to 2.5, altered values of regional circulation, increased II up to 3-4 points one should diagnose average degree of process flow. At increased level of UK, HIS being above 0.75 mcM/ml/h, F-1-P, L-SD, L-TD, LP products, CIC by 2 and more times, increased IRI being above 2.5, altered values of regional circulation, increased II up to 5 points and more one should diagnose severe degree of process flow.
EFFECT: higher accuracy of diagnostics.
FIELD: medicine, infectology, hepatology.
SUBSTANCE: in hepatic bioptate one should detect products of lipid peroxidation (LP), such as: dienic conjugates (DC), activity of antioxidant enzymes, such as: catalase (CAT)and superoxide dismutase (SOD). One should calculate by the following formula: C = DC/(SOD x CAT)x100, where DC - the content of dienic conjugates, SOD - activity of superoxide dismutase, CAT - activity of catalase. At coefficient (C) values being above 65 one should predict high possibility for appearance of cirrhosis, at 46-645 - moderate possibility and at 14-45 -low possibility for appearance of cirrhosis.
EFFECT: higher accuracy of prediction.
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: clinical medicine, pulmonology.
SUBSTANCE: one should carry out complex estimation of interleukin-1β) concentration in blood, saliva, bronchoalveolar liquid. Moreover, one should detect distribution coefficient (DC) for IL-1β as the ratio of IL-1β blood content to IL-1β salivary content. At increased IL-1β blood content by 10 times and more, by 2 times in saliva, unchanged level of bronchoalveolar IL-1β, at DC for IL-1β being above 1.0 one should predict bronchial obstruction. The method enables to conduct diagnostics of the above-mentioned disease at its earlier stages.
EFFECT: higher efficiency of prediction.
FIELD: medicine, diagnostics.
SUBSTANCE: the present innovation deals with genetic trials, with diagnostic field of oncological diseases due to analyzing DNA by altered status of gene methylation that take part in intracellular regulation of division, differentiating, apoptosis and detoxication processes. One should measure the status of methylation in three genes: p16, E-cadherine and GSTP1 in any human biological samples taken out of blood plasma, urine, lymph nodes, tumor tissue, inter-tissue liquid, ascitic liquid, blood cells and buccal epithelium and other; one should analyze DNA in which modified genes of tumor origin or their components are present that contain defective genes, moreover, analysis should be performed due to extracting and purifying DNA out of biological samples followed by bisulfite treatment of this DNA for modifying unprotected cytosine foundations at keeping 5-methyl cytosine being a protected cytosine foundation followed by PCR assay of bisulfite-treated and bisulfite-untreated genes under investigation and at detecting alterations obtained according to electrophoretic result of PCR amplificates, due to detecting the difference in the number and electrophoretic mobility of corresponding fractions at comparing with control methylated and unmethylated samples containing normal and hypermethylated forms of genes one should diagnose oncological diseases. The method provides higher reliability in detecting tumors, detection of remained tumor cells after operation.
EFFECT: higher efficiency of therapy.
1 cl, 3 dwg, 4 ex
FIELD: medicine, gastroenterology.
SUBSTANCE: one should carry out diagnostic studying, moreover, on the 5th -6th d against the onset of exacerbation in case of gastric and duodenal ulcerous disease one should detect the content serotonin, histamine and acetylcholine in blood, then during 2-3 wk one should conduct medicinal therapy to detect serotonin, histamine and acetylcholine level in blood again and at serotonin content being by 2-3 times above the norm, histamine - by 1.15-1.4 times above the norm and acetylcholine - by 20-45% being below the norm one should predict the flow of gastric and duodenal ulcerous disease as a non-scarring ulcer.
EFFECT: higher accuracy of prediction.
SUBSTANCE: method involves taking blood from ulnar vein (systemic blood circulation) and from large vein of the injured extremity proximal with respect to lesion focus (regional blood circulation). Spontaneous NST-test value is determined and difference is calculated in systemic and regional blood circulation as regional-to-systemic difference. The difference value is used for predicting clinical course of pyo-inflammatory disease in extremities.
EFFECT: high accuracy of diagnosis.
4 cl, 2 tbl
FIELD: medicine, gastroenterology.
SUBSTANCE: one should introduce biologically active substance, moreover, in patient's blood serum one should detect the content of acetyl choline and choline esterase activity followed by 2-h-long intragastric pH-metry at loading with biologically active substance as warm 40-45%-honey water solution at 35-40 C, and at increased content of acetyl choline being above 1.0 mM/l, choline esterase being above 0.5 mM/l/30 min and pH level being 6.0-6.9 it is possible to consider apitherapy to be useful for treating ulcerous duodenal disease.
EFFECT: higher efficiency and accuracy of detection.
FIELD: medicine, gastroenterology.
SUBSTANCE: it has been suggested a new method to detect pharmacological sensitivity to preparations as acidosuppressors. After the intake of the preparation a patient should undergo fibrogastroduodenoscopy 3 h later, then, through endoscopic catheter one should introduce 0.3%-Congo red solution intragastrically and the test is considered to be positive at keeping red color that indicates good sensitivity to the given preparation, and in case of dark-blue or black color the test is considered to be negative that indicates resistance to this preparation. The suggested innovation widens the number of diagnostic techniques of mentioned indication.
EFFECT: higher efficiency of diagnostics.