RU2338191C1 - Method of endogenous intoxication severity evaluation in newborn with hyperbilirubinemia - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention refers to medicine, namely to paediatrics and infectious diseases, and can be used for endogenous intoxication severity evaluation in newborn with hyperbilirubinemia. Substance of method: capillary blood in amount 0.02 ml is sampled and tested for erythrocyte membrane permeability by osmotic erythrocyte resistance. If erythrocyte membrane permeability is 12.05±0.06% slight endogenous intoxication is diagnosed. Erythrocyte membrane permeability 29.8±0.015% shows medium endogenous intoxication, and if this value is 58.23±0.24%, severe endogenous intoxication is diagnosed.

EFFECT: higher accuracy of endogenous intoxication severity evaluation at newborn hyperbilirubinemia.

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Description

The invention relates to medicine, namely to pediatrics and infectious diseases, and can be used to assess the severity of endogenous intoxication in children with hyperbilirubinemia.

Endogenous intoxication (EI) is a non-specific syndrome characteristic of many diseases. The prognosis of the course of diseases, the choice of a method of detoxification therapy is impossible without an objective assessment of the degree of endogenous intoxication. To date, many methods for diagnosing EI have been developed. The most objective of them are: the definition of a humoral agent responsible for the development of EI - the definition of the so-called intoxication marker. But we can distinguish too large a group of toxic substances of endogenous origin (bilirubin, bile acids, residual nitrogen fractions, circulating immune complexes, and so-called medium-sized molecules or molecules of medium weight (MSM) and an even larger group of exogenous toxic factors). Determining MSM is a rather expensive method and requires certain equipment. Methods for diagnosing EI by testing the patient’s blood on biological agents require certain working conditions and considerable time. Thus, the problem of intoxication marker is still far from the final solution. In most cases, the fact of the discovery of a toxic agent is not enough to adequately explain the clinical data. Detoxification mechanisms in each organism have an individual functional threshold. In practical activities, the doctor needs to determine the degree of intoxication in a matter of minutes and start resuscitation measures in a timely manner, the outcome of treatment often depends on the timeliness of the beginning of which.

In recent years, interest has increased in studying the mechanisms of the involvement of the blood system in the body's response to stress and extreme effects. Red blood cells are the first to come into contact with toxic agents, primarily due to their multiplicity and ability to change their shape and form aggregates. Contact with toxic agents damages red blood cell membranes. Thus, damage to erythrocyte membranes can be considered as an integral indicator of the toxicity of exo and endotoxins.

As a prototype, the authors propose a "Method for the diagnosis of endogenous intoxication syndrome" V. Lazoreva, patent N RU 2234095 from 08/10/2004, g, including an assessment of the permeability of erythrocyte membranes in the presence of an aqueous solution of NaCl in dilutions, the number of red blood cells is determined in the Goryaev’s chamber and with the number non-hemolyzed red blood cells, 10 times the control, diagnosed with endogenous intoxication syndrome.

Along with the undoubted advantages, the method has some disadvantages:

1) for this method, venous blood is used in an amount of 4 ml - 5 ml, which is not acceptable for pediatric practice (especially in neonatology);

2) the method ascertains the very fact of the presence or absence of endogenous intoxication in patients, without characterizing individual degrees of EI in hyperbilirubinemia, in particular;

3) the age of the subjects from 17 to 76 years - this is a significant scatter, which may affect the results of the determination of TEM;

4) patients taken for examination, with multiple organ pathology, which may also affect the results of the determination of TEM;

5) the blood of children has not been tested, and in children many biochemical processes proceed differently from adults due to the fact that children develop and form adaptation systems (nervous, immune and endocrine).

The technical result of the claimed inventors is to increase the speed and accuracy of determining the severity of endogenous intoxication with hyperbilirubinemia in newborns. It is proposed: 1) micromethod (0.02 ml of blood is enough); 2) the quantitative method (more accurate and faster: the degree of hemolysis was determined not in the Goryaev’s chamber, which is subjective, but on the FEK, by the concentration of free hemoglobin).

Red blood cells are diluted with saline in a ratio of 1: 2 (0.04 saline is added to 0.02 ml of capillary blood) and 0.01 ml of suspension are placed in 5 centrifuge tubes, 2.5 ml of working solution are added to each centrifuge tube increasing concentration of urea. The scheme for the preparation of working solutions for the determination of TEM is shown in table 1.

Table 1
The scheme of preparation of working solutions for the determination of TEM Urea solution 0.3 mol / l (ml) NaCl solution 0.87% (ml) The concentration of urea in the working solution (mol / l) 35 65 0.11 fifty fifty 0.15 55 45 0.165 60 40 0.18 65 35 0.195 one hundred 0 0.3

Hemolysis in mixtures of isotonic solutions of urea and sodium chloride is due to the ability of urea to quickly diffuse through the cell membrane and, creating a hyperosmolar concentration inside the red blood cell, cause it to swell, followed by hemolysis. The degree of hemolysis is expressed in units of optical density. TEM is defined as the ratio of the optical density of the sample to the optical density of the sample with 100% hemolysis (6th test tube), multiplied by 100%. The degree of hemolysis is proportional to the permeability of this population of red blood cells.

It was experimentally established that the most informative for the determination of TEM were the optical density of hemolysis with a working solution of 60:40 (4th test tube).

The material for the study was the capillary blood of 96 newborns from 0 to 8 months., Who were in the intensive care unit of the children's hospital No. 1 in Tver, taken as planned to monitor their condition. It was found that normal (clinically healthy newborns) erythrocyte membrane permeability (TEM) is 5.04 ± 0.2%. TEM indices in newborns with hyperbilirubinemia had a wide scatter. With a mild degree of hyperbilirubinemia, the TEM indices amounted to 12.05 ± 0.06%, with moderate severity - 29.8 ± 0.015%, and in severe form - 58.23 ± 0.24%.

The results of the study are shown in table 2.

table 2
TEM indices of dependence on the stage of endogenous intoxication (EI) No. Measurable indicators Control n = 13 Light degree of EI n = 28 Moderate degree of EI n = 34 Severe degree of EI n = 21 one The number of red blood cells (million / ml) 5.3 ± 0.26 5.3 ± 0.27 4.8 ± 0.24 4,5 ± 0,23 2 The degree of hemolysis (units of optical density) 0.5 ± 0.026 1.6 ± 0.79 1.98 ± 0.01 2.5 ± 0.1 3 TEM (erythrocyte membrane permeability)% 5.04 ± 0.2 12.05 ± 0.06 29.8 ± 0.015 58.23 ± 0.24

Clinical trials of "a method for determining the severity of endogenous intoxication with hyperbilirubinemia in newborns" showed that the method allows to predict the degree of endotoxemia with a forecast accuracy of 85%. Diagnostically significant for the transition to a new state (healthy, that is, no intoxication - mild - moderate - severe severity of intoxication) is an approximately twofold increase in TEM (5.04 ± 0.2-12.05 ± 0.06-29, 8 ± 0.015-58.23 ± 0.24). There is a kind of qualitative leap.

Clinical example No. 1

Patient S., 18 days old, case history 1409, was admitted to the neonatal pathology unit of the Children's City Hospital No. 1 for severe anxiety (monotonous crying), slight yellowness of the skin and sclera, liver +3 cm, hyperthermia up to 38 degrees C. PEM was determined immediately upon admission (result within 30 minutes: red blood cells in a child 4.7 Giga \ l, the degree of hemolysis in the experimental sample (60:40) = 1.393 TEM = Eop. / E (100% heme.) * 100% = 1.393 / 2,401 * 100% = 58,017%, which indicated a severe degree of endogenous intoxication). The patient was transferred to the intensive care unit, where he received intensive care. The final diagnosis: hemoliquorodynamics disturbance, stage 3 hyperbilirubinemia, severe. The forecast has been fully confirmed.

Clinical example No. 2

Patient M., 2 months old, medical history 1287, was admitted to the neonatal pathology unit of the Children's City Hospital No. 1 due to severe sclera jaundice, body temperature 38 degrees C, vomiting once with a fountain, liver +2 cm. TEM was determined immediately upon admission (result within 30 minutes: red blood cells 5.1 Giga \ l, the degree of hemolysis in the experimental sample (60:40) = 0.1201, TEM = 0.1201 / 1.001 * 100% = 11.99%). A mild degree of endogenous intoxication was diagnosed. After detoxification measures, the child was transferred to the infectious diseases department. By the end of the first day of hospitalization, the temperature returned to normal, the child is active, sucks well, the stool is normal, there was no vomiting, and the yellowness of the sclera decreased significantly. Final diagnosis: ARVI: enterovirus infection, stage 1 hyperbilirubinemia, mild. The forecast was confirmed.

Clinical example No. 3

Patient D., 4.5 months, medical history 1187, was admitted to the intensive care unit of the city hospital No. 1 due to yellowness of the sclera and palms, repeated vomiting of the fountain, dehydration (sunken fontanel), and elevated body temperature up to 38.5 degrees C. Liver +3 cm. The chair is light. The boy’s mom is on the 8th day in the infectious diseases unit for hepatitis A. Grandmother was sitting with the child. According to the grandmother, he fell ill acutely. Lack of lethargy, fever, vomiting. Ambulance delivered to the hospital. On admission, a TEM was determined in a child (result within 30 minutes: erythrocytes 4.7 Giga \ L, degree of hemolysis in the experimental sample (60:40) = 0.58, TEM = 0.59 / 1.98 * 100% = 29.798 %).

A moderate severity of intoxication was diagnosed. After detoxification measures, the child was transferred to the infectious diseases department a day later. The final diagnosis: hepatitis A, stage 2 hyperbilirubinemia, moderate to severe severity. The forecast has been fully confirmed.

Claims (1)

A method for determining the severity of endogenous intoxication in newborns with hyperbilirubinemia, including a blood test, characterized in that they take 0.02 ml of capillary blood, determine the permeability of erythrocyte membranes by the osmotic resistance of red blood cells, and when the permeability of erythrocyte membranes is 12.05 ± 0, 06% diagnose a mild degree of endogenous intoxication, with a value of 29.8 ± 0.015% - diagnose a moderate degree of endogenous intoxication, and with a value of 58.23 ± 0.24% - a severe degree of endogenous intoxication.