Differential diagnostic technique for hypoxia type

FIELD: medicine.

SUBSTANCE: blood plasma is analysed for cytochrome oxidase activity, erythrocyte 2,3- biphosphoglycerate and lactic acid concentrations. It is followed by calculating an oxygenation coefficient K by formula: K=(C1+C2):A, where C1 is the erythrocyte 2,3- biphosphoglycerate concentration, mol/l; C2 is the erythrocyte lactic acid concentration, mol/l; A is plasma cytochrome oxidase activity, mol/l. The K value withint 1.0 <K≤3.0 enables to diagnose tissue hypoxia, while in case of the K values being 3.0<K≤5.0 cardiovascular hypoxia is diagnosed. If the oxygenation coefficient is 5.0 and more, blood hypoxia is diagnosed.

EFFECT: use of the method enables the differential diagnostics of blood, tissue and cardiovascular hypoxia.

3 ex

 

The invention relates to medicine and can be used for differential diagnosis of the type of hypoxia in various pathologies in cardiology, obstetrics and gynecology, dermatology and other medical fields.

Hypoxia - a condition that occurs when the mismatch between the needs of the cells in oxygen delivery, or when compliance is achieved as a result of excessive voltage activity oxygen transport system of the organism (Spector VA Hypoxia. // The Bulletin of intensive therapy, 2007, No.1, p.12-15).

"Etiological factor" distinguish 4 types of hypoxia: respiratory, blood, tissue, and cardiovascular (Great medical encyclopedia Academy of medical Sciences of the USSR. - M., 1966, s-923.).

Respiratory type of hypoxia caused by reduced partial pressure of oxygen in the air we breathe, due to various environmental factors, such as low atmospheric pressure, raising the ambient temperature, etc. that reduce the diffusion of oxygen to the tissues.

Tissue type of hypoxia due to impaired absorption of oxygen by the tissues and occurs when the poisoning, violate tissue respiration (connection hydrocyanic acid), avitaminosis, hormonal deficiency.

Cardiovascular type of hypoxia on is due to the reduction of blood pressure in diseases of the heart and blood vessels, when the shock and collapse, which leads to a decrease in blood oxygen saturation.

Blood type hypoxia due to impaired transport of oxygen to the cell and occurs when a blood disease, bleeding, hypoplastic bone marrow.

Regardless of the type of hypoxia can lead to reduced mental and physical performance, incoordination, headache, nausea, disturbance to realistically assess ambient, slow speech, and in severe cases, convulsions, decreased blood pressure, respiratory arrest, loss of consciousness and death of the organism (Zbarskaya B. I., Ivanov I.I., Murdashev S.R. Biological chemistry. - L., Medicine, 1972, s; Shevchenko UL Hypoxia: adaptation, pathogenesis, clinic. - SPb.: LLC ALBI-SPb", 2000, p.12).

Differential diagnosis of respiratory type of hypoxia is based on the analysis of the data of the anamnesis of a patient's life and the peculiarities of its stay in places of high altitude and usually does not cause difficulties. Differential diagnosis of tissue, cardiovascular and blood types of hypoxia, based only on clinical symptoms, causes considerable difficulties.

In this regard, the development of new methods for differential diagnosis of the type of hypoxia based on the study of the mechanisms of its development is one of the urgent problems of health (Simonen the s A.P., Fedorov V.D., Kluyev VM Specification classification hypoxic conditions. // Bulletin of the Russian Academy of medical Sciences, 2004, No. 2, p.46), as the tactics of examination and management of patients should be carried out taking into account the specific type of hypoxia.

Conducted research on the medical-scientific and patent literature has identified various ways to diagnose the type of hypoxia.

Author's certificate of the USSR No. 1215657 (1986, BI No. 9), a protected Way to diagnose violations of oxygen exchange" consisting in the determination of the partial pressure and the rate of its utilization in the ischemic tissue. The study was conducted on polarography in constant current mode, so the cathode is a platinum electrode open type, is injected into the muscle of the forearm to a depth of 15-20 mm from the skin surface, and the anode - silver chloride electrode, placed on the back surface of the brush. Source current count value of the oxygen tension in the tissue, and then create an ischemia and to determine the rate of oxygen consumption, and after removal of ischemia - speed ratio of oxygen reduction. On the basis of the received data calculated oxygen balance of the fabric and when it is <1.0 to diagnose the tissue type of hypoxia.

The disadvantage of this method is that the process of its implementation causes certain difficulties, such as the presence of the equipment and trained personnel. This method is technically complex and requires the creation of regional ischemia tissue, which in some cases can worsen the General condition of the patient. In addition, this method is not sufficiently informative, because you can only determine the tissue type of hypoxia without differential diagnosis of other types of hypoxia.

In the patent of Russian Federation №2155533 (2000, TPMF No. 25) described "a Method for the diagnosis of disorders of the use of oxygen in acute poisoning". To implement this method, the patient carried out the sampling of arterial blood and mixed blood, as well as the fence mixed exhaled air. Using the standard analyzer to measure blood pH, gas composition of blood samples and air actual and expected consumption of oxygen expect a deficit of oxygen. If there is a deficiency of oxygen greater than 25, then diagnose the tissue type of hypoxia.

The disadvantage of this method is the low information content, because it can be diagnosed only tissue type of hypoxia. Furthermore, the method is technically challenging, since its implementation requires the simultaneous analysis not only of samples of arterial blood and mixed blood, but also the analysis of breath samples.

RF patent №2241378 (2004, BIPM No. 34) protected "Method of determining the degree of utilization of sour what kind of tissues in the body", which consists in determining the value of oxygen saturation of arterial and mixed venous blood with subsequent calculation of the coefficient of utilization of oxygen. The measurement of oxygen saturation of arterial blood and mixed blood carried out by the method of reflective spectrofotometry, for which the catheter fiber optic sensors mounted simultaneously in the femoral and pulmonary artery. The obtained value of the coefficient of oxygen utilization compared with the normal value, and when it is below the norm diagnose cardiovascular type of hypoxia.

The disadvantage of this method is its technical complexity and lack of information, because only allows you to diagnose cardiovascular type of hypoxia.

The closest technical solution chosen as a prototype, is protected by copyright certificates of the USSR (No. 1673041, 1991, IB No. 32) "Method for the diagnosis of hypoxia, which is the study of blood of the patient. In the arterial blood of the patient determine the amount of hemoglobin and oxygen before and after saturation of oxygen saturator under standard conditions. Then by the formulas calculate the integral indicator of oxygen deficiency and compared with normal values. If its normal value exceeds the value by 3-5% - diagnose respiratory type of hypoxia, and if its size exceeds the normal value by more than 5% are diagnosed with a blood type of hypoxia.

The disadvantage of the prototype is its complexity and difficulty, as it is necessary to use special equipment for oxygen saturation additional portion of the arterial blood in vitro, which requires the presence of a specially trained personnel. In addition, this method is not very informative, because it allows to diagnose only respiratory and blood types of hypoxia.

The aim of our invention is to simplify and expand the functionality of the method.

This goal is achieved by the fact that exploring the venous blood of the patient, in the plasma which depends on the activity of cytochrome C oxidase A. In erythrocytes of venous blood to determine: the concentration of 2,3-diphosphoglycerate C1and the concentration of lactic acid2. Then using the formula to calculate the value of the coefficient oxygenation (K):

K=(C1+C2):A,

where:

C1the concentration of 2,3-diphosphoglycerate in erythrocytes (mol/l);

With2- the concentration of lactic acid in the blood (mol/l);

And activity of cytochrome C oxidase in the plasma (mol/l).

If the value K is in the range of 1.0<K≤3,0, diagnose tissue type of hypoxia. If the value of the coefficient of the oxygenation To lie in the range of 3.0< To≤5,0, diagnose cardiovascular type of hypoxia. When the value of the ratio of oxygenation To>5,0, diagnose blood type of hypoxia.

Detailed description of the method.

The patient takes a 4.5 ml of blood from a vein in the centrifuge tube, add 0.5 ml of heparin. Then the sample is centrifuged in a centrifuge at 3000 rpm for 10 minutes. In the centrifugation isolated plasma and erythrocytes.

In the resulting plasma determine the activity And cytochrome C oxidase by a spectrophotometric method for Krivchenkov R.S. (Krivchenkov R.S. determination of the activity of cytochrome C oxidase in the suspension of mitochondria. Modern methods in biochemistry. M., 1977, p.47-49) in the reaction with dimethyl-para-phenylalanine at a wavelength of 536 nm. Activity is expressed in mol/L.

In erythrocytes obtained by the colorimetric method to determine the concentration of C12,3-diphosphoglycerate according to the method of Lagunovoi ACTING and M. Blinov. (Lukanova I.S., M. Blinov. Determination of 2,3-FGD non-enzymatic method and ATP in erythrocytes of patients with chronic lymphocytic leukemia. Laboratory business. 1975, No. 7, s-654). Calculated by a calibration curve obtained results are expressed in mol/L.

The concentration of C2lactic acid in the blood venous blood was determined by the colorimetric method by Menshikov V.V. (Menshikov V.V. Laboratory methods of research the Oia at the clinic. M., 1987, s). Calculated by a calibration curve, the results are expressed in mol/L.

Then calculate the ratio of oxygenation To the formula:

K=(C1+C2):A,

where:

C1the concentration of 2,3-diphosphoglycerate in erythrocytes (mol/l);

With2- the concentration of lactic acid in the blood (mol/l);

And activity of cytochrome C oxidase in the plasma (mol/l).

If the value K is in the range of 1.0<K≤5,0, diagnose tissue type of hypoxia. If the value of the ratio of oxygenation To lie in the range of 3.0<K≤5,0, diagnose cardiovascular type of hypoxia. When the value of the ratio of oxygenation To>5,0, diagnose blood type of hypoxia.

Practical feasibility of the proposed method is illustrated by examples from clinical practice.

Example 1: patient M, 49, case history No. 2006 went to the polyclinic №6 of Rostov-on-don in the office of the dermatologist with a preliminary diagnosis: onychodystrophy. The patient complained of stratification and thinning of the nail plates hand, the presence of longitudinal grooves on their surface. Diagnosis - onychodystrophy confirmed. In an interview with the patient, it was also revealed that she suffers from dizziness and fatigue with little exertion. When examination revealed clinical signs of hypoxia: pallor of agnihotramv, hypothermia, acrocyanosis and to determine the type of hypoxia, the patient survey was conducted according to the claimed method.

The patient took from Vienna to 4.5 ml of blood from a vein in the centrifuge tube was added 0.5 ml of heparin. The obtained blood was centrifugally at the Central laboratory centrifuge-1 at 3000 rpm for 10 minutes. In the centrifugation allocated plasma and erythrocytes.

In the obtained plasma was determined activity And cytochrome C oxidase by a spectrophotometric method for Krivchenkov R.S. (Krivchenkov R.S. determination of the activity of cytochrome C oxidase in the suspension of mitochondria. Modern methods in biochemistry. M., 1977, p.47-49) in the reaction with dimethyl-para-phenylalanine at a wavelength of 536 nm. The activity of cytochrome C oxidase, A=13.5 mol/L.

In the resulting erythrocytes identified C1the concentration of 2,3-diphosphoglycerate according to the method of Lagunovoi I.S. and M. Blinov. (Lukanova I.S., M. Blinov. Determination of 2,3-FGD non-enzymatic method and ATP in erythrocytes of patients with chronic lymphocytic leukemia. Laboratory business. 1975, No. 7, s-654). The value of C1=8.1 mol/L.

In erythrocytes was determined by the concentration of C2lactic acid colorimetric method for Menshikov V.V. (Menshikov V.V. Laboratory methods in the clinic. M., 1987, s). The value of2=10.4 mol/L.

Then calculated the ratio of oxygenation To the formula:

K=(C1+C2):And=(8,1+10,4):13,5=1,4.

As the value of the coefficient oxygenation K=1,4 is in the range of 1.0<K≤3,0, the patient was diagnosed with tissue type of hypoxia and prescribes medication. The results of the treatment and no complications have confirmed the reliability of the delivered according to the claimed method of diagnosis.

Example 2: patient N., 29, the case history No. 18926 was admitted to the maternity ward MUSL GB No. 1 of Rostov-on-don with the preliminary diagnosis of placental insufficiency, preeclampsia. The diagnosis of placental insufficiency, preeclampsia has been confirmed. The patient complained of weakness, dull pain in the chest, shortness of breath with little exertion. When examination revealed clinical signs of hypoxia: pale skin, acrocyanosis, shortness of breath, and the violation of uteroplacental blood flow and to determine the type of hypoxia, the patient survey was conducted according to the claimed method.

The patient took from Vienna to 4.5 ml of blood from a vein in the centrifuge tube was added 0.5 ml of heparin. The obtained blood was centrifugally at the Central laboratory centrifuge-1 at 3000 rpm for 10 minutes. In the centrifugation allocated plasma and erythrocytes.

In the obtained plasma was determined activity And cytochrome C oxidase with antropologicheskii method on Krivchenkov R.S. (Krivchenkov R.S. determination of the activity of cytochrome C oxidase in the suspension of mitochondria. Modern methods in biochemistry. M., 1977, p.47-49) in the reaction with dimethyl-para-phenylalanine at a wavelength of 536 nm. The activity of cytochrome C oxidase And=3.8 mol/L.

In the resulting erythrocytes determined the concentration of C12,3-diphosphoglycerate according to the method of Lagunovoi I.S. and M. Blinov. (Lukanova I.S., M. Blinov. Determination of 2,3-FGD non-enzymatic method and ATP in erythrocytes of patients with chronic lymphocytic leukemia. Laboratory business. 1975, No. 7, s-654). The value of C1=5.7 mol/L.

In erythrocytes was determined by the concentration of C2lactic acid colorimetric method for Menshikov V.V. (Menshikov V.V. Laboratory methods in the clinic. M., 1987, s). The value of2=10.4 mol/L.

Then calculated the ratio of oxygenation To the formula:

K=(C1+C2):And=(5,7+10,4):3,8=4,2.

As the value of the ratio of oxygenation To=4,2 lies in the range of 3.0<K≤5,0, the patient was diagnosed with a cardiovascular type of hypoxia and prescribes medication. The results of the treatment and no complications have confirmed the reliability of the delivered according to the claimed method of diagnosis.

Example 3: patient M, 43 years old, medical history, No. 8202 addressed in the gynecological Department of the Rostov state what about the medical University with a preliminary diagnosis: uterine fibroids, complicated by bleeding. The patient complained of bleeding, heavy menstruation for 2 weeks. Diagnosis of uterine fibroids, abnormal bleeding, was confirmed. When examination revealed clinical signs of hypoxia: pale skin, acrocyanosis, cyanosis of nasolabial triangle, shortness of breath, tachycardia, decreased blood pressure 100/75 mm Hg and to determine the type of hypoxia, the patient survey was conducted according to the claimed method.

The patient took from Vienna to 4.5 ml of blood from a vein in the centrifuge tube was added 0.5 ml of heparin. The obtained blood was centrifugally at the Central laboratory centrifuge-1 at 3000 rpm for 10 minutes. In the centrifugation allocated plasma and erythrocytes.

In the obtained plasma was determined activity And cytochrome C oxidase by a spectrophotometric method for Krivchenkov R.S. (Krivchenkov R.S. determination of the activity of cytochrome C oxidase in the suspension of mitochondria. Modern methods in biochemistry. M., 1977, p.47-49) in the reaction with dimethyl-para-phenylalanine at a wavelength of 536 nm. The activity of cytochrome C oxidase And=3.1 mol/L.

In the resulting erythrocytes determined the concentration of C12,3-diphosphoglycerate according to the method of Lagunovoi I.S. and M. Blinov. (Lukanova I.S., M. Blinov. Determination of 2,3-FGD non-enzymatic method and ATP in erythrocytes Bo is lnyh chronic lymphocytic leukemia. Laboratory business. 1975, No. 7, s-654). The value of C1=14,85 mol/L.

In erythrocytes was determined by the concentration of C2lactic acid colorimetric method for Menshikov V.V. (Menshikov V.V. Laboratory methods in the clinic. M., 1987, s). The value of2=to 4.38 mol/L.

Then calculated the ratio of oxygenation To the formula:

K=(C1+C2):And=(14,81+4,38):3,1=6,2.

As the value of the ratio of oxygenation To=6,2>5, the patient was diagnosed with a blood type of hypoxia, and prescribes medication. The results of the treatment and no complications have confirmed the reliability of the delivered according to the claimed method of diagnosis.

We examined 112 patients, including 20 pregnant women with placental insufficiency, 32 patients with onychodystrophy, 60 patients with uterine myoma. All patients had clinical signs of hypoxia and to determine the type of hypoxia them a survey was conducted according to the claimed method. The survey found that 40 patients had blood type hypoxia, 34 patients had tissue type of hypoxia, 38 patients had cardiovascular type of hypoxia. The diagnosis was confirmed by the results of the treatment.

Thus, the claimed "a Method of differential diagnosis of the IPA hypoxia" is easy to implement and allows for differential diagnosis of blood, tissue and cardiovascular types of hypoxia.

The method of differential diagnosis of the type of hypoxia, which consists in the study of blood, wherein investigate venous blood plasma which determine the activity of cytochrome C oxidase And, in erythrocytes is the concentration of 2,3-diphosphoglycerate C1and the concentration of lactic acid2then calculate the ratio of oxygenation K according to the formula:
K=(C1+C2):A,
where C1the concentration of 2,3-diphosphoglycerate in red blood cells, mol/l;
With2- the concentration of lactic acid in the blood, mol/l;
And activity of cytochrome C oxidase in the plasma, mol/l,
and if the value of K lies in the range of 1.0<K≤3,0, diagnose tissue type of hypoxia, if the value of K lies in the range of 3.0<K≤5,0, diagnose cardiovascular type of hypoxia, when K>5,0 diagnose blood type of hypoxia.



 

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4 ex

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.

3 ex

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