SU1642395A1 - Method for diagnosis of hyperthymism in children - Google Patents

Abstract

The invention relates to practical medicine, in particular to clinical immunology and pediatrics, can be used in practical health care when examining children with suspected hyperplasia of the thymus gland and their further treatment with the use of immunomodulators. The purpose of the invention is to simplify the method, reduce the time and reduce the complications. For this, an immunological examination of children is carried out and with an increase in the absolute number of lymphocytes to 6.5-8.5, the absolute number of T-lymphocytes to 2.5 4,0 "1 O9 / l and B lymphocytes up to 1.2-2.6 10 / l or an imbalance of the relative and absolute content of T and B lymphocytes, which consists in reducing the relative number of T lymphocytes to 15-30% with normal values of their absolute number and increased the absolute number of lymphocytes up to 6.5 8.5 "109 / l and B-lymphocytes up to 1.2-2, diagnosed ie the presence of Megalith timo- children. S to

Description

The invention relates to practical medicine, in particular to clinical immunology and pediatrics, can be used in practical public health when examining children with suspected hyperplasia of the thymus gland and their further treatment using immunomodulators.

The purpose of the invention is to simplify, reduce the time and likelihood of complications.

The method is carried out as follows.

DGH implementation of the method, you must calculate the total number of leukocytes, lymphocytes and determine the number of T-and B-lymphocytes in the blood.

Determining the number of T- and lymphocytes.

1. Blood collection Blood is obtained from a fingerless finger. After ethanol treatment, the finger is punctured with a scarifier and blood in a volume of 0.2 ml is gravity-fed into a tuberculin syringe (pre-moistened with a solution of heparin with a concentration of 200 units / ml and dried at 37 ° C in a thermostat). The blood is placed in a I0 ml glass centrifuge tube and shaken thoroughly.

2. The selection of material for research. The number of leukocytes and

About 4

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Lymphocytes are determined by standard procedures, for which 0.02 ml of blood is taken. The remaining blood is centrifuged at 200 g for 1 min. The exfoliated plasma is poured, and the sediment is used to isolate leukocyte suspension.

3 ". Preparation of leukocyte suspension. Leukocyte suspension is obtained by lysis of erythrocytes with a hemolytic fluid of the following composition; 0.04 g / l of EDTA was added to a 0.083% ammonium chloride solution. The pH of the solution is adjusted to 7.2 "7.4 7; sodium bicarbonate solution. To the blood cells left after centrifugation, 5 ml of hemolytic fluid is added for 30 seconds while stirring constantly with Yath then ml of medium 199 is added to stop the hemolytic effect. cells per cent, centrifuged with iO min at 200 g, the supernatant was poured: the precipitate was gently shaken. 1-2 ml of hemolytic liquid was added for 3-5 seconds, then 10 ml of medium 199 was added, 0 was centrifuged min at 200 g. The supernatant is drained. The remaining liquid is removed with filter paper. To the resulting cell pellet, 0.2 ml of medium 199 "is added.

4J T-cell count and lymphocyte counting, Lymphocyte T-B cell count is carried out using the rosette pattern. The reactions are set up in 96 well plates for single-use immunological reactions.

To the well of the tablet, 0.05 ml of the obtained leukocyte suspension is added, 0905 ml of a sheep erythrocyte suspension is added to determine T-lymphocytes or 0.05 ml of a 0.5% mouse erythrocyte suspension to determine B-lymphocyte The cap on the plan is fixed and centrifuged at 200 g for 5 minutes. The plate is incubated for 30 minutes at ° C, then a drop of 3% glutaraldehyde solution is carefully added to the wells, kept at room temperature for 5 minutes. After that, the supernatant is removed, and 0.1 ml of distilled water is poured into the wells. The sediment is carefully re-suspended and smears are prepared in

5 Determination of the absolute content of rosetting cells in 1 μl of blood

ttys / mkd -r ° L%

ROL

thousand / mm

100%

five

where O is one thousand / μL the absolute number of rosetting

10 lymphocytes in 1 μl

blood;

five

THO / ICA

ROL%

- absolute lymphocyte content

1 µl blood;

- the number of rosettes - forming lymphocytes,% a

A total of 369 children aged from four months to four 0 years were examined. Of these, 52 people had normal T and B lymphocytes, the absolute values of lymphocytes were significantly higher than normal.

The content of lymphocytes in the blood de 5 TI with timomegaly are given in Tables e

High absolute indices of T and B lymphocytes, an increase in the absolute value of lymphocytes characterize their functional immaturity due to thymomegaly. Thus, the absolute number of lymphocytes in children with thymomegaly is increased to 6.5-8, the absolute number of T-lymphocytes to 2.5-4.0 10 / L, and B-lymphocytes 1.2-2. In addition, there may be an imbalance between the relative and absolute number of lymphocytes, which consists in the relative number of T lymphocytes up to 15-30% with normal values of their absolute and elevated absolute numbers lymphocytes up to 6.5 8, and B-lymphocytes up to 1.2-2.6 09Agi

Example 1, a patient with HC, 2 da6 A child entered the nursing ward of a children's regional hospital with a diagnosis of pneumonia, prolonged

five

0

Radiology of a child

revealed: the median tissue in the upper and middle third of the chest is dilated in both directions due to the shadow of the thymus CTT-index 0} 58

An immunological examination revealed: lymphocytes 8, / l, T-ROL 52%, abs. 4.22 CH09 / L, V-ROL 38%, abs. 2.81 109 / l Increase in absolute lymphocyte counts, and

In addition, T and B-Mn cells also indicate their functional immaturity and is due to thymomegaly, which was confirmed by an X-ray ™ nevsky examination

Example 2 “Patient K,

1year 3 months ", Case History No. 335 Entered the skin department of Children's Hospital No. 3 with diagnosis: detskaya eczema

An immunological examination of the patient revealed: Lymphocytes 7.94 109 / L; T-ROL 52%, abs. 4.13 x / 109 / l, V-ROL 20%, abs. , 59 109 / l, a pronounced increase in absolute lymphocyte counts, and B-ROL indicates a functional immaturity of lymphocytes characteristic of thymomegaly, which was confirmed by X-ray (CTT-index 0.45),

Example Sick S,,

2 years old, Entered the skin department of Children's Hospital No. 3 with a diagnosis of detskaya eczema,

An immunological examination established that: lymphocytes 8.6-10 / T-ROL 8%, absp 1.55-109 / l, B-ROL 15%, abs. 1.29-109 / l The apparent imbalance between relative and absolute indicators of T and B lymphocytes is due to functional non-maturity of lymphocytes, revealed during thymomegaly. However, during x-ray examination of a patient in direct projection, the thymus thymus shadow was not determined and KGT index did not exceed 0} 38 "When X-ray examination in the lateral projection, an increase in the thymus gland

Analyzing the data presented, it can be concluded that the changes in the display

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the body’s immunlotus status in the form of a significant increase in T and B lymphocyte counts, as well as an imbalance between relative and absolute indicators with an increased absolute number of lymphocytes, can serve as a basis for making a timomegali diagnosis.

ten

The use of the proposed method for the diagnosis of timomegaly in children has the following advantages: there is no need to carry out X-ray

15 physical examination of children; reduction of research time and complexity (research time is reduced to 4 hours; no R-setting, film is needed); with mass immunological examinations

20 dr. Children gives the opportunity to identify children with timomegaly and carry out timely preventive treatment; there is no side ,, effect.

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Claims (1)

Invention Formula Method for the diagnosis of timomegaly in children by clinical research ZS Vani, distinguished by the fact that, in order to simplify, reduce time and reduce complications, a blood test is conducted to determine the number of lymphocytes of T and B lymphocytes and 3f with lymphocyte count (6, 5-8.5) 10 cells / l, absolute number of T-lymphocytes (2.5-4.0) -I O9l / l with normal values of the relative number or relative number 4C T-lymphocytes 15-30% with a normal absolute number, the number of cells (1, 2-2.6) -10 cells / l diagnose the presence of thymomegaly in children.