Method for predicting disorders in lipid exchange system in children

FIELD: medicine.

SUBSTANCE: the present innovation deals with testing and analyzing biological materials and could be applied for predicting disorders in lipid exchange system in children. One should apply a drop of urine onto a slide and cover it with a cover glass. Due to keeping constant environmental conditions the preparation should be kept till the appearance of pronounced typical structures on a slide. The preparation should be tested due to testing the whole surface of the sample in light field followed by polarization-optic survey. Testing results should be fixed. If there are no textures on a slide in light field and there are a great number of small double refracting lamellar and acicular crystals simultaneously it is possible to diagnose no disorders in lipid exchange system. If there are geometric dendrites and atypical crystals in light field on a slide simultaneously, and in polarized light one should simultaneously observe insignificant quantity or large double refracting lamellar and acicular crystals, spherolytes, lens-like crystals it is possible disorders in lipid exchange system should be diagnosed. The method excludes invasiveness and simplified the technique suggested.

EFFECT: increased accuracy of diagnostics.

2 ex

 

The invention relates to medicine, in particular to the study or analysis of biological materials, and can be used for the diagnosis of dysfunctions of lipid metabolism in children.

There is a method of determining the dysfunctions of lipid metabolism, including children, in accordance with which the conduct biochemical study of blood lipid spectrum by determining the amount of total cholesterol, lipoproteins of high and low density, triglycerides, and atherogenic index (Ministry of health guidelines of the USSR /edited by Angelova and Yeganian, 1975).

The disadvantage of this method lies in the complexity of the qualitative and quantitative determination of the above components and the large role of the subjective factor in obtaining this information. This reduces its validity, since in practice this leads to a discrepancy between the clinical symptoms, morphological substrate and lipid spectrum. From practice it is known that even in patients with severe clinical manifestation of the disease in approximately 1/3 of cases recorded normal levels of the individual components of lipid metabolism.

Furthermore, the method does not allow you to diagnose early stages of the disease, and diagnose the presence or absence of illness, because each criterion is valid is the limit, above which speaks of the violation in the system of lipid metabolism.

The disadvantage of this method is invasive, it does not exclude the possibility of infection in the patient's body.

Closest to the present invention is a method of determining dysfunctions of lipid metabolism, including children (A.S. USSR №1723527, G 01 N 33/92, 30.03.92), in accordance with which the patient take venous blood, extracted total lipids and register their content. In this extract total lipids crystallized on a slide, and then register the type of texture in polarized light. Upon detection of texture myelin forms and fan crystals detect the violation in the system of lipid metabolism.

The disadvantage of this method consists in the following. Use as an investigational biological substances serum complicates the known method and reduces its efficiency due to the necessity of preparation of the drug for research and because of the complexity of its preparation. This storage of the drug is not desirable. This is because unlike most components of laboratory specimens, in which the storage at low temperatures or in dried condition significantly increases the stability, the lipoproteins of human serum. It says Otomi, preparation serum should be investigated as soon as possible, because the time delay reduces the reliability of the data used for diagnostics. In addition, upon receipt of serum unexpected may be affected by the coagulation factor. For example, in the serum, not fully coagulated, formed of strands of fibrin or clot, sometimes within a few hours after the end of the process. This reduces the reliability of the method.

The disadvantage of this method is also invasive, that does not exclude the possibility of infection in the patient. In addition, the process of collecting venous blood from a patient, especially a child affects his mental state, puts it in a stressful situation, which in some cases may affect the diagnostic results, especially for patients suffering from diseases of the cardiovascular system.

Thus, the detected analog and the prototype of the present invention when the implementation does not achieve the technical result consists in increasing the reliability, the exception invasiveness, to simplify the method and to increase its efficiency.

The present invention solves the task of creating a method for the diagnosis of dysfunctions of lipid metabolism, the implementation of the ie which allows to achieve a technical result, consisting in the increase of reliability, the exception invasiveness, to simplify the method and to increase its efficiency.

The essence of the invention lies in the fact that in the method for the diagnosis of dysfunctions of lipid metabolism in children, including morphological study of the biological fluid by examining its texture liquid crystal structure in the dynamics in polarized light, the analysis of research results and the fact of the presence or absence of abnormalities in the system of lipid metabolism, in addition, the texture of the liquid crystalline structure of the biological fluid is studied in the light box, and as a biological fluid using the urine, which on the surface of the glass put a drop of urine and covered with a cover glass, and then, while maintaining the environmental conditions constant, the product can withstand, until on the subject the glass expressed typical textures, followed by a study of the drug by a visual inspection of the entire surface of the sample in a light box, and then produce a polarization-optical study of the drug, the results of the inspection record, if on a slide in the bright field not see the texture, and in polarized light at the same time see a large number of small birefringent plate and needle-shaped crystals, t is diagnosed in the absence of disturbances in the system of lipid metabolism, if on a slide in bright field at the same time see geometric dendrites and atypical crystals in polarized light at the same time see a small amount or large birefringent lamellar and acicular crystals, spherulites, lenticular crystals, diagnose deficiencies in the system of lipid metabolism.

The technical result is achieved in the following way. Many liquid biological environment of the human organism is able to crystallize, and in certain conditions - to move into the intermediate liquid state. In the liquid-crystalline state of the environment, while maintaining fluidity, manifests in polarized light of a specific crystalline pattern - texture. It is known that the biological fluid are multicomponent systems, most of them showing structural heterogeneity (heterogeneity) and high sensitivity to the composition and the form of existence of the components. The composition of the liquids adequately reflects the physiological state of an organism, as well as the functional usefulness of its various organs and systems. For example, regulatory mechanisms and pharmacological factors affect the quantitative content of protein and calcium salts in the urine, the ratio of saturated and unsaturated lipids in CIV is the pigs blood, the nature of the aggregation of the lipid complex of bile, the number of phospholipids, derivatives of cholesterol and its esters exhibiting liquid crystalline properties. These changes in thin molecular level are manifested, in particular, features of the aggregation of biological fluids at the level of microstructures. Morphology textures of liquid crystalline phases correlated with body condition and changes in the presence of pathology, which allows us to observe that the dynamics in the bright field and polarized light in a conventional optical magnification (A.S. USSR №1209168, And 61 In 10/00, 07.02.86; A.S. USSR №1486932, G 01 N 33/92, 15.06.89; A.S. USSR №1723527, G 01 N 33/92, 30.03.92; RF patent №2173462, G 01 N 33/48, 33/68, 10.09.2001; RF patent №2170432, G 01 N 33/48, 33/68, 10.07.2001).

In the proposed method for the diagnosis of dysfunctions of lipid metabolism in children use morphological study of the biological environment, namely urine. Due to the fact that urine can crystallize, passing through an intermediate liquid state, it is possible morphological examination of the urine by studying its texture liquid crystal structure in the dynamics of bright-field and polarized light by a visual inspection of the entire surface of the sample.

In the proposed method for studies prepare the drug from the urine, which on the surface of the slide cause is aplu urine and covered with a cover glass. Due to the fact that the drug serves a cover glass, allows the diagnosis of dysfunctions of lipid metabolism by examining the texture of liquid crystal structures of the investigated drops of urine. This is explained in the following. It is known that lipids are different in their chemical nature of the substance. The lipids are neutral fats - triglycerides, cholesterol (total, free and bound), phospholipids (lipids containing as a mandatory component phosphorus), and glycolipids (Vsemirnov "Clinical laboratory tests from a to z and their diagnostic profiles", Minsk: Belarusian Nauka, 1999, s). Self-organization of fluids during dehydration has a fairly clear pattern. First of all, it will be determined by the separation of salts and organic substances, which is explained by different degrees of hydrophilicity salt and organic substances. The drop has the form of a hemisphere, which has a different thickness in the center and the periphery. As a result, in blatant cover glass a drop of urine in the evaporation of water is uneven changes in the concentration of dissolved substances, namely: concentration in thin peripheral areas is increasing at a faster pace compared to the Central (thick) part drops. In these conditions manifests itself hydrophilic dissolved biojidkosti components. The first to begin to crystallize the salt. Salt, in the fight for the remaining water, they quickly move to the center of the drop, "squeezing" of organic matter on the periphery. Therefore, in the formation of typical textures in the outdoor drop of urine is dominated by salt crystals. On their background texture of organic substances subtle and not very informative.

A drop of urine, covered with a cover glass, the crystallization processes occur otherwise. In this case, the liquid is distributed on the glass evenly. Because the drug is covered by a cover glass and the environmental conditions constant support, when the evaporation of water is also uniform change in the concentration of dissolved substances. Furthermore, the molecules of organic and inorganic substances are not pushed out to the periphery, and are allocated between the part of the urine salt molecules, taking an active part in the formation of typical base textures. This allows us to reliably detect not only the presence in the drop of urine organic and inorganic substances, but their quantitative and qualitative changes, and therefore increases the reliability of the results of diagnostics of malfunctions in the system of lipid metabolism.

Education expressed a typical basic textures on a slide means the end of the aggregation process. what then makes it inappropriate to further increase the exposure time of the drug determines the start time for the study of the texture.

In the claimed invention the study of the preparation can be done by a visual inspection of the entire surface of the sample in a light box, and then produce a polarization-optical study of the drug, the results of the inspection record. Inspection of surface preparation twice: in the bright field and polarized light allows you to get complete information about the texture of the investigated drops of urine, because some of the crystal structure occur only in polarized light.

If on a slide in the bright field not see the texture, and in polarized light at the same time see a large number of small birefringent plate and needle-shaped crystals, then diagnose the absence of disturbances in the system of lipid metabolism.

If on a slide in bright field at the same time see geometric dendrites and atypical crystals in polarized light at the same time see a small amount or large birefringent lamellar and acicular crystals, spherulites, lenticular crystals, diagnose deficiencies in the system of lipid metabolism.

Experienced by the inventors proved that unsaturated lipids included in the composition of the studied biological environment - urine, the formation of texture is rolled in the form of a small plate crystals, which manifest that the are under polarized light and are birefringent. It is known that a healthy child's body is characterized by a high level of metabolism. Therefore, in healthy children, in which the compensatory ability of the kidneys high amounts of unsaturated lipids in the urine more than patients. This explains the greater number of lamellar crystals in the texture of the investigated drops of urine of healthy children, in comparison with sick children, and therefore, allows you to diagnose the violation or no violation in the system of lipid metabolism. In the normal presence in the texture of the urine in large amounts in the aggregate only small plate and needle-shaped birefringent crystals indicates the absence of anomalies in the qualitative and quantitative composition of the investigational drops of urine.

The appearance in the texture of the investigated drops of urine abnormal crystals: bright field - of atypical crystals and geometric lead, polarized-light - spherulites and lenticular crystals, evidence of qualitative changes in the composition of urine, is not characteristic of the composition of urine in norm. Reducing the number and increasing size of the plate and needle-shaped birefringent crystals also tells about the deviation from the norm qualitative and quantitative composition of the investigational drops of urine.

From the above it follows that the study of the dynamics of liquid crystalline structure of the tours of the investigated liquid - urine bright field and polarized light allows high accuracy to diagnose deficiencies in the system of lipid metabolism, since the texture of the liquid crystalline structure of the investigated drops of urine reflects the dynamics of currency in the system of lipid metabolism. It gives a complete picture of the qualitative and quantitative composition of urine at this time. In this examination on a slide texture investigated drops of urine in the bright field and polarized light allows you to get complete information about the morphology of the investigated drops of urine, because some of the crystal structure occur only in polarized light, which also increases the reliability of diagnostic results. In addition, it proves the possibility of using urine as the study of the biological environment, to diagnose deficiencies in the system of lipid metabolism. The result is a non-invasive way. Noninvasive method makes it safe for the patient against infection infection and has no emotional impact. The latter is especially important when working with children. Furthermore, the reduced risk of contamination of personnel through the study medication, as, unlike urine, blood preparation (in our case, serum) may be a carrier of AIDS.

The use of urine as the e of the investigated biological fluid simplifies the claimed method of diagnosis, unlike the prototype, which is used as a biological fluid is serum, as the preparation process of the preparation for the study, the proposed method is reduced to the fence urine of a patient and the location of the investigated drops of urine between the objective and the cover glass. The ease of preparation of the drug for research simplifies the method and increases its efficiency.

Easy sampling of biological fluids for research and the lack of need for special preparation of the drug for research increases the efficiency of the proposed method in comparison with the prototype.

Thus, the present invention "Method for the diagnosis of dysfunctions of lipid metabolism in children during implementation ensures the achievement of the technical result consists in increasing the reliability, the exception invasiveness, to simplify the method and to increase its efficiency.

In addition, the present invention in comparison with the prototype, achieving additional technical result consists in the possibility of assessing the compensatory ability of the kidneys. This is because in the proposed method, lipid metabolism is controlled by exploring the biological fluid is urine, which is the product of work by the EC and its structure adequately reflects their functional state. If this violation of lipid metabolism controlled by quantitative changes in the content of unsaturated lipids in the urine, which are the product of the kidneys. Because the authors have proved empirically that unsaturated lipids included in the composition of the studied biological environment - urine, the formation of texture is rolled in the form of a small plate crystals, the texture reflects the number of unsaturated lipids in the drop of urine at this time. This allows analyzing the nature of the texture of urine, to ascertain not only the absence or disruption of lipid metabolism, but also to assess the compensatory ability of the kidneys, namely: the more the texture is fine lamellar crystals (unsaturated lipids), the compensatory ability of the kidneys better.

The claimed method for the diagnosis of dysfunctions of lipid metabolism in children is as follows. As a biological fluid using the urine. Spend a morphological study of the biological fluid by examining its texture liquid crystal structure in the dynamics of bright field and polarized light. On the surface of the glass put a drop of urine and covered with a cover glass. Then, while maintaining the environmental conditions constant, the product can withstand up appeared which I slide expressed typical textures. After conducting a study of the drug by a visual inspection of the entire surface of the sample in a light box, and then produce a polarization-optical study of the drug. The results of the inspection record. If on a slide in the bright field not see the texture, and in polarized light at the same time see a large number of small birefringent plate and needle-shaped crystals, then diagnose the absence of disturbances in the system of lipid metabolism. If on a slide in bright field at the same time see geometric dendrites and atypical crystals in polarized light at the same time see a small amount or large birefringent lamellar and acicular crystals, spherulites, lenticular crystals, diagnose deficiencies in the system of lipid metabolism.

In all examples, the execution method for the preparation of drugs from urine were taken pre-processed subject and cover glasses. On the glass put a drop of urine, then covered with cover glass. The drug was sustained, maintaining constant environmental conditions, to appear on the surface of glass slides of typical textures.

In all examples, the execution method for the preparation of drugs from urine were taken pre-processed subject article is Klah. Pay attention to the quality of processing slides, in order to avoid artifacts in the research. A glass slide was washed with distilled water, then degreased by immersion in 96% of medical alcohol and wipe dry in the same direction dry sterile cloth.

View bright field produced when divorced Nikolaj with increasing h... × 250. Inspection of the entire surface of the sample was made longitudinally transverse scanning with a step equal to the magnitude of the field of view.

View in polarized light produced when crossed Nikolaj with increasing h... × 250. Inspection of the entire surface of the sample was made longitudinally transverse scanning with a step equal to the magnitude of the field of view. All detected features were fixed.

Research can be used microscopes series BIOLAM (with polarized filters, FLOORS, MBI.

For a small number of crystals took the case when the crystals take no more than 20% of the area of the field of view at 150-fold increase and no more than 2 from 5-7 fields of view.

For the small size of the crystals took the case when the crystal is in1/4the field of view and takes less than 0.1 from him.

Example 1. Patient K., age 7, examination.

When the survey closed drops of urine in the bright field texture is not observed. When you view the e in polarized light in a closed drop of urine are observed simultaneously in a large number of small birefringent needle-shaped crystals and lamellar crystals.

Diagnosis: lack of dysfunctions of lipid metabolism.

Example 2. Patient C., age 8, examination.

When the survey closed drops of urine in the bright field in the drug detected geometric dendrites and atypical crystals of various shapes. When viewed in polarized light in the drug at the same time there are large birefringent lamellar and acicular crystals in a small amount, and birefringent spherulites and lenticular crystals.

Diagnosis: the presence of disturbances in the system of lipid metabolism in this patient.

The diagnostic results are confirmed by conventional laboratory tests.

Method for the diagnosis of dysfunctions of lipid metabolism in children, including morphological study of the biological fluid by examining its texture liquid crystal structure in the dynamics in polarized light, the analysis of research results and the fact of the presence or absence of abnormalities in the system of lipid metabolism, characterized in that, in addition, the texture of the liquid crystalline structure of the biological fluid is studied in the light box, and as a biological fluid using the urine, which on the surface of the glass put a drop of urine and covered with a cover glass, and then, while maintaining the environmental conditions constant, p is apart stand until you see slide expressed typical textures after conducting a study of the drug by a visual inspection of the entire surface of the sample in a light box, and then produce a polarization-optical study of the drug, the results of the inspection record, if on a slide in the bright field not see the texture, and in polarized light at the same time see a large number of small birefringent plate and needle-shaped crystals, then diagnose the absence of disturbances in the system of lipid metabolism, if on a slide in bright field at the same time see geometric dendrites and atypical crystals in polarized light at the same time see a small amount or large birefringent lamellar and acicular crystals, spherulites, lenticular crystals then diagnose deficiencies in the system of lipid metabolism.



 

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