RU2475747C1 - Method of predicting psychomotor development of children with perinatal affection of central nervous system - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: invention relates to problem of impairment of psychomotor development in children and can be applied for assessment of prediction of psychomotor development retardation in children with perinatal affection of central nervous system (PA CNS) of hypoxic-ischemic genesis in anamnesis. Activity of leukocyte elastase is determined, after that additionally spectrophotometrically activity of α1-protease inhibitor is determined and levels of LE and α1-PI activity are assessed comprehensively, and combination of initial LE level >225 nm/min/ml and initial level of α1-PI activity >30 IU/ml testifies to positive prediction of psychomotor children development. Combination of initial LI level >225 nm/min/ml and initial level of α1-PI activity <30 IU/ML testifies to negative prediction of psychomotor children development.

EFFECT: application of claimed method makes it possible to use immunologic parameters: level of LE activity and level of α1-protease inhibitor activity, as objective criteria for assessing prediction of psychomotor development dynamics in children.

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Description

The invention relates to medicine and relates to the problem of psychomotor development disorders in children and can be used to diagnose and predict the prognosis of psychomotor development delay in children with a perinatal lesion of the central nervous system (CNS) of a history of hypoxic-ischemic origin.

At an early age, various terms are used: "neuropsychic development", "psychomotor development", "mental development", which are identical for a given age period (Big Medical Encyclopedia).

Recent studies have convincingly demonstrated the involvement of the immune system in the development of various forms of central nervous system pathology, including those caused by impaired development of the nervous system. It was revealed, in particular, the participation of autoimmune reactions in the process of impaired brain development. This is evidenced by the identification in the blood of children of the first year of life with brain dysginesis and mental dysontogenesis of a high level of autoantibodies to the nerve growth factor - a protein of brain tissue that performs neurotrophic functions. It is very significant that the level of autoantibodies to the nerve growth factor correlates with the characteristics of the clinical status of patients. At the same time, the state and role of nonspecific (innate) immunity in cases of psychomotor developmental disorders remain poorly understood. However, it is known that the system of acquired and innate immunity is closely related, and the activation of one of these systems (in this case, autoimmune reactions) indicates certain changes in the system of innate immunity

Currently, the only methods for assessing the delay in psychomotor development (ZMPR) of young children are clinical, neuro-psychological methods that are based on the use of psychomotor development scales developed at different times: the “Gnome” scale (Kozlovskaya G.V., Goryunova A.V. ., Samokhvalova VI The methodology for determining the mental development of children under 3 years of age - GNOME // Journal of Neurology and Psychiatry, 1997; 97: 8: p. 38-42), "Development Tables" by A. Gesell, 1925, N. scales. Bailey, 1969, Denver Scale, 1973, Munich Functional Diagnostics, 1985, the scale proposed by L.G. Zhurba and E.M.Mastyukova, 1981, the scale developed by O.V. Bazhenova, 1986.

The relationship between the nervous and immune systems, neuroimmunological parallels and the dynamics of the neuropsychiatric status of the child can be traced by immunological indicators (Thesis "The formation of psychomotor functions and prediction of deviations in the development of deeply premature children in the 1-2 year of life". Author: Sakharova, Elena Stanislavovna, k. M.S., 2003).

A known method for predicting a delay in neuropsychiatric development in young children with perinatal CNS lesions by examining a child aged 1 year, according to which in the blood serum of a child with a normal level of neuropsychic development, the content of ceruloplasmin is determined and when its value is 20-41, 6 mg / dl, predict the IV-V group of neuropsychic development by 3 years of life (RF patent No. 2338203).

A known method for predicting perinatal hypoxic lesions of the central nervous system in newborns by immunological examination of the peripheral venous blood of a pregnant woman, while from the 35th week of pregnancy complicated by gestosis, the content of interleukin-1 is determined, and with its value> 40 pcg / l, the development of perinatal hypoxic lesions of the central nervous system (RF patent No. 2313095).

A known method for determining the activity of leukocyte elastase in blood plasma, including diluting the blood plasma with physiological saline, adding a buffer solution and substrate to it, followed by spectrophotometric determination of the rate of cleavage of the substrate, while the blood plasma is diluted 15 to 20 times, using Tris HCI, and as a substrate Vos Ala-O-Np, diluted in acetonitrile (RF Patent No. 2039983).

There is a method of determining the nature of pathophysiological disorders of the psychomotor development of children of the first year of life, according to which blood is taken, serum is separated, enzymatic and enzyme immunoassay are performed, and the activity of leukocyte elastase (LE) and autoimmune reaction are determined by the level of antibodies to nerve growth factor (NGF), with an increase in LE activity and the absence of an autoimmune reaction to NGF, perinatal encephalopathy is detected, while a simultaneous increase in LE activity and the level of autoantibodies to NGF are evidenced t about organic damage to the brain (RF Patent No. 2231074).

The method we have chosen as a prototype.

The objective of the invention is to develop a new method for predicting psychomotor development in children with perinatal damage to the central nervous system.

The technical result of the task is to select objective criteria for assessing the dynamics of the psychomotor development of children with perinatal damage to the central nervous system.

The essence of the invention lies in the fact that at the same time, the parameters of innate immunity are determined spectrometrically: the level of activity of the leukocyte elastase and the α1-protease inhibitor in peripheral blood. Then, the activity levels of LE and α1-PI are comprehensively assessed, while the combination of the initial level of LE> 225 nm / min / ml and the initial level of activity of α1-PI> 30 IE / ml indicates a positive prognosis of the psychomotor development of children with ZPMR and Central nervous system hypoxic a history of ischemic origin. The higher the activity indicator of LE and the lower the activity indicator of the α1-protease inhibitor, the more pronounced the delay in the psychomotor development of this category of children is indicated.

Leukocyte elastase (LE) - a proteolytic enzyme, a marker of the functional activity of neutrophils, is released into the extracellular space due to activation by various factors of an infectious, and most importantly, endogenous nature. In perinatal lesions of hypoxic-ischemic genesis, activators of the innate immune response are ligands of endogenous origin. By endogenous factors we mean cell breakdown products (necrosis and apoptosis). These immunopathological parameters play a key role in the development of secondary metabolic disorders in the brain with perinatal damage to the central nervous system of hypoxic-ischemic origin.

In close relationship with LE is its antagonist - the αl-protease inhibitor (α1-PI) - a glycoprotein synthesized by the liver. This protein controls the proteolytic activity of LE, the functional activity of α1-PI determines the course of many inflammatory and / or destructive processes. The lack of activity of this protein causes uncontrolled activation of LE and the inflammatory process associated with secondary metabolic damage to body tissues. The level of functional activity of α1-PI reflects the state of the compensatory potential of blood serum.

The method is as follows. The parameters of innate immunity are determined spectrometrically: the level of activity of leukocyte elastase (LE) and the level of activity of its antagonist, an α1-protease inhibitor (α1-PI). To do this, they take blood from children, separate the serum, dilute it with physiological saline, add a buffer solution and substrate to it, followed by spectrophotometric determination of the rate of cleavage of the substrate using the ELASTAZA and ALFA 1-PI reagents kit, manufactured by Biofarm Test LLC ", Moscow. Plasma separated from the cell pellet was diluted 15-20 times with physiological saline, added to a spectrophotometer cuvette, Tris-HCl buffer warmed up to 28-30 ° C was added, and then Boc-Ala-O-Np substrate dissolved in acetonitrile was added. After incubation, an increase in the optical density of the sample is recorded at a wavelength of 347.5-410.0 nm for 8-10 minutes and the enzyme activity is calculated using a specific formula.

In the SCCH RAMS, 90 patients aged 1-12 months with a delay in psychomotor development and perinatal CNS damage of a hypoxic-ischemic genesis in the anamnesis were examined: 70 full-term babies and 20 premature babies.

In 50 children, psychomotor development was re-evaluated after 6-12 months and the level of leukocyte elastase and α1-protease inhibitor in the blood serum was determined.

The average level of LE in the blood of healthy children from the comparison group, presented as: M (normal) ± SD (standard deviation), is 175 ± 25 nm / min / ml.

The LE level of 225 nm / min / ml was chosen as a criterion for dividing children into groups due to the fact that it is 175 + 2SD nm / min / ml.

The average activity of α1-PI in the blood of healthy children from the comparison group, presented as M ± SD, is 32.5 ± 2.5 IU / ml.

The value of α1-PI activity, equal to 30 IE / ml, was chosen by us as a criterion for dividing children into groups due to the fact that it is 32.5 - SD.

During the verification of this assumption, it was found that in the group of children with the initial level of LE> 225 nm / min / ml and the initial activity index α1PI (> 30 IE / ml), the specific gravity of children with an improvement in the MTCT score of 20% or more was significantly higher than in the group of patients with the same initial LE level, but with a low α1PI activity index (<30 IE / ml).

The data obtained indicate that the level of LE> 225 nm / min / ml in combination with normal and high activity of α1-PI (α1PI> 30 IE / ml) can be regarded as a favorable prognostic factor in the dynamics of psychomotor development of children with perinatal damage to the central nervous system .

Clinical examples of the method

Example 1. Boy A., age - 5 months. The main diagnosis: The consequences of perinatal damage to the central nervous system. Hyperactivity Syndrome. A disorder of the autonomic autonomic nervous system. Concomitant diagnosis: Hypotrophy of the 1st degree. Anemia of the 1st degree. Bilateral hydrocele. Examined by a neurologist, conducted a psychological and pedagogical examination. A moderate delay in psychomotor development was revealed: the total MTCT score on the Gnome scale was 50. By the spectrophotometric method using the ELASTASA and ALFA 1-PI reagents, we determined the indicators of innate immunity: leukocyte elastase (LE) level and its antagonist level - α1 protease inhibitor (α1-PI). All determinations were carried out in blood samples taken from a finger, heel, or from a vein into dry plastic tubes. After the formation of a blood clot, a blood sample was centrifuged at 2.5 thousand rpm, serum was taken, which was used for analysis. The child was examined in the dynamics of psychomotor development after 8 months. He was again consulted by a neurologist and a psychological and pedagogical examination was conducted. Revealed compensation ZPMR. The total MTCT score during the second examination on the “Gnome” scale was 90.

Table 1 presents the indicators of innate immunity in this patient during the initial and repeated examination.

Table 1 Indicators PMR score Indices of innate immunity LE activity level (nm \ min \ ml) The level of activity of α1-PI (IE / ml) Primary examination fifty 315 52 Re-examination (after 8 months) 90 201.4 34.5 Comparison Group (healthy children) 95-100 175 ± 25 32.5 ± 2.5

The presented clinical case demonstrates good dynamics of psychomotor development, i.e. full compensation for a moderate delay in psychomotor development, associated with a significant drop in the level of LE and normalization of α1-PI activity in a child with an initially moderate ZPMR and central nervous system PP with a history of hypoxic-ischemic origin.

Example 2. Girl A., age 7 months. The main diagnosis: The consequences of perinatal damage to the central nervous system. The syndrome of muscle dystonia hypertonic type. Prematurity 32 weeks. Examined by a neurologist, conducted a psychological and pedagogical examination. A gross delay in psychomotor development was revealed: the total PMR score on the Dwarf scale is 25. The indicators of innate immunity are estimated. The child was examined again after 11 months. He was again consulted by a neurologist and a psychological and pedagogical examination was conducted. A gross delay in psychomotor development has persisted. The total MTCT score during the second examination on the “Gnome” scale was 30.

Table 2 presents the indicators of innate and acquired immunity in this patient during the initial and repeated examination.

table 2 Indicators PMR score Indices of innate immunity LE activity level (nm / min / ml) Activity level
α1-PI (IE / ml) Primary examination 25 240 eighteen Re-examination (after 11 months) thirty 268 36 Comparison Group (healthy children) 95-100 175 ± 25 32.5 ± 2.5

The presented clinical case demonstrates the absence of dynamics of psychomotor development, i.e. the child retained a gross delay in psychomotor development, associated with an increase in the level of LE and activity of α1-PI in a child with an initially severe ZPMR and central nervous system PP of a history of hypoxic-ischemic genesis and an increased level of LE and at the same time a decreased activity of α1-PI in blood serum compensatory potential).

The proposed method allows the use of immunological indicators: the level of activity of LE, the level of activity of the α1-protease inhibitor as objective criteria for assessing the prognosis of the dynamics of psychomotor development of children with a delay in psychomotor development and perinatal damage to the central nervous system with a history of hypoxic-ischemic origin.

Claims (1)

A method for predicting the psychomotor development of children with perinatal damage to the central nervous system, including determining the activity of leukocyte elastase (LE), characterized in that the activity of the α1-protease inhibitor (α1-PI) is additionally determined spectrophotometrically, then the activity levels of LE and α1-PI are comprehensively assessed, the combination of the initial level of LE> 225 nm / min / ml and the initial level of activity of α1-PI> 30 IE / ml indicates a positive prognosis of the psychomotor development of children, and the combination of the initial level E> 225 nm / min / ml and the initial level of activity of α1-PI <30 IU / ml indicates a negative prognosis psychomotor development of children.