Method for prediction of clinical course of autoimmune thyroiditis
SUBSTANCE: peripheral venous blood of the patients with autoimmune thyroiditis is examined for the absolute transferring receptor (CD71+) T-lymphocyte count and the interleukin-2 (IL-2) concentration. If the CD71+ T-lymphocyte count exceeds 0.3×109 cell/l, and the IL-2 concentration is less than 25 pg/ml, a favourable clinical course of the disease is predicted, while the CD71+ T-lymphocyte count less than 0.3×109 cell/l and the IL-2 concentration exceeding 25 pg/ml shows an unfavourable clinical course of autoimmune thyroiditis.
EFFECT: method provides more accurate and objective prediction with using the material, reagents and research methods widely applied in common clinical-laboratory practise.
The invention relates to immunology, endocrinology, laboratory diagnosis, and is intended for prediction of clinical course of autoimmune thyroiditis.
Autoimmune thyroiditis (AIT) suffer from 3-4% of the world's population, with age, the number of patients increases, while the prevalence in women is 3 to 6 % per year. Recently, clinicians in many countries the increasing number of patients with autoimmune thyroiditis among young people, especially among children and adolescents (thyroid Disease, Ed. of Bravermann LI - M.: Medicine, 2000). In General, the analysis of blood in patients with autoimmune thyroiditis possible lymphocytosis, monocytosis, leukopenia and increased ESR. Immunological blood test allows to detect the increase in the number and activity of T-helper and T-killer cells by reducing the number of T-suppressor cells, increased levels of immunoglobulins.
There is a method of evaluating the secretion of TSH, which includes sample with thyroliberin. The thyrotropin-releasing hormone stimulates the release of TSH thyroid-stimulating cells adenogipofiza, so normal after the introduction of thyroliberin TSH level increases. Sample with thyroliberin used to assess secretion of TSH reserve and the degree of suppression of TSH secretion (thyrotoxicosis). Response to thyrotropin-releasing hormone reduced or delayed amid suppressive therapy levotiroxina recent sample with thyroliberin rarely used, because of the new highly sensitive methods for the determination of TSH (Endocrinology edited Nlevine. - M., 1999).
There is a way to evaluate the effectiveness of treatment of AIT, including the definition in the serum of patients contents of interferon-gamma (Patent No. 2323443, Russia. Publ. 27.04.2008). After 1 month of treatment with the concentration of interferon-gamma less than 35 PG/ml appreciate the treatment as effective, and when the content is more than 35 PG/ml evaluate treatment as ineffective and adjust the dose of therapeutic drug or change the medication.
Also known is a method for predicting and diagnosing diseases of the thyroid gland (US 2010/0131286 A1, 27.05.2010), which investigated the level of expression of specific marker genes, indicating that the risk of the development and distribution of endocrine pathologies, including autoimmune thyroiditis. In the specified method is used, however, expensive equipment and reagents, which is not always economically feasible.
The prototype is to study the serum levels of cytokine BLyS, stimulating b-lymphocytes for the diagnosis, prognosis and screening the effectiveness of therapy of autoimmune diseases (WO 2009043848, 09.04.2009). The method includes comparing the concentration of BLyS with a reference value and the subsequent calculation of variances for the purpose of diagnosis, prognosis and screening. However, the content of titoki is and BLyS serum increases not only when AIT, systemic lupus erythematosus, rheumatoid arthritis and sjögren's syndrome, but also in patients with multiple myeloma, chronic b-cell leukemia that is not sufficiently objective criterion for predicting the nature of the flow AIT. We stated the invention improves the accuracy and objectivity of forecasting, using two criteria: the absolute content of T-lymphocytes with the transferrin receptor (CD71+) and the concentration of interleukin-2 (IL-2). When the content of lymphocytes CD71+ more than 0.3×109CL/l and the concentration of IL-2 less than 25 PG/ml predicts favorable course of the disease, and when the content of lymphocytes CD71+ less than 0.3×109CL/l and the concentration of IL-2 for more than 25 PG/ml predict unfavorable for AIT. Used in the way materials, reagents and methods are widely used in standard clinical and laboratory practice.
The claimed method is standard in the following way. Blood should be taken from the cubital vein in a minimal volume (1 ml) to 10 o'clock in the morning, on an empty stomach. The content of T-lymphocytes CD71+ were determined using indirect immunoperoxidase reactions with the use of monoclonal antibodies for the preparations of lymphocytes type "dried droplet" production RPC Medispectra Moscow. For the quantitative determination of IL-2 is used serum or PLA is mA blood analyzed by ELISA with the use of a diagnostic kit for ELISA. The results of the reaction are measured on a spectrophotometer with a wavelength of 450 nm.
In this way were examined persons rehabilitated in a medical company "Biochar" Arkhangelsk: 28 patients with autoimmune thyroiditis and 15 people have a course of hormone treatment of thyroid cancer and are in remission (control). Diagnosed by the doctors of the city endocrinology Advisory centre of Arkhangelsk, the survey was carried out in compliance with the basic norms of biomedical ethics. The results were processed using the software package Statistica 6" ("StatSoft, USA). Study type retrospective, random sample, simultaneously. General population - residents of the North of European Russia. The significance of differences between groups was evaluated using a parametric t-student test for independent samples and non-parametric criterion of Wilcoxon signed. Statistical significance was assigned at p<0,05.
It is established that under unfavorable course of the disease (increase in blood concentrations of antibodies to thyroperoxidase) patients the decrease of the absolute content of T-lymphocytes CD71+ from 0.47±0,05 to 0.26±0,02×l09to the/l and increased concentrations of IL-2 23,8±5.3 to 154,5±11,7 compared to controls (p< 0,05). Under favorable for AIT (decrease in the level of antibodies to thyroperoxidase) increases the content of lymphocytes CD71+ to 0.56±0,05×109CL/l and normalized index of IL-2 (less than 25 PG/ml).
There is information on the stimulating effect of thyroxine on the release of soluble alpha-receptors to IL-2 (Zwirska-Korczala K. Influence of thyroxine on serum soluble interleukin-2 receptor alpha levels in thyroid disorders / K. Zwirska-Korczala, A. Berdowska, J. Jochem et al. // J. Clin. Pharm.Therapeut. - 2004. - Vol. 29, N 2. - R. 151-156), the latter can neutralize the excess concentration of IL-2 and thereby reduce the level of autoimmune inflammation in the thyroid gland. It is known that IL-2 and transferrin, are powerful enough proliferative potential (Zinkernagel R. fundamentals of immunology: TRANS. with it. - M.: Mir, 2008. - 135 C.). At the same time, given that transferrin supports cell proliferation, and blockade of receptors for transferrin in transparentbikini cells induces in them apoptosis, reduction in the blood concentration of CD71+ cells can be considered as a compensatory mechanism in the regulation of proliferative process (Neckers L. Geldanamycin as a potential anti-cancer agent: its molecular target and biochemical activity/ L.Neckers, T.W.Schulte, E.Mimnaugh // Investigational New Drugs. - 1999. - V.17, No. 4. - S-373).
Examples of use of the method.
1. Patient P., aged 51 years. Diagnosis: autoimmune thyroiditis, confirmed high levels of antibodies to thyroperoxidase 56 IU/ml The patient received treatment with L-thyroxine in a dose of 100 mcg/day. During the course of hormonal therapy in peripheral venous blood of the patient set the absolute content of T-lymphocytes CD71+ 0,24×109CL/l, the level of IL-2 were 286 PG/ml Made a forecast of unfavorable course of the disease, which is confirmed by the further increase in the concentration of antibodies to thyroperoxidase.
2. Patient W., age 54 years. Diagnosis: autoimmune thyroiditis, confirmed high levels of antibodies to thyroperoxidase 176 IU/ml the Patient is assigned to treatment with L-thyroxine in a dose of 75 mcg/day. During the course of hormonal therapy in peripheral venous blood of the patient set the absolute content of T-lymphocytes CD71+ 0,22×109CL/l, the level of IL-2 was 89 PG/ml Made a forecast of unfavorable course of the disease, which is confirmed by the further increase in the concentration of antibodies to thyroperoxidase.
3. Patient B., aged 54 years. The diagnosis of autoimmune thyroiditis with a characteristic increase in the level of antibodies to thyroperoxidase 186 IU/ml the Patient is assigned to treatment with L-thyroxine in a dose of 50 mg/day. During the course of hormonal therapy in peripheral venous blood of the patient set the absolute content of T-lymphocytes with the transferrin receptor (CD71+) of 0.62×109CL/l, the level of IL-2 was 17.3 PG/ml the forecast has been Made favorable for the it, that is confirmed by further reducing the concentration of antibodies to thyroperoxidase.
A method for predicting the clinical course of autoimmune thyroiditis, characterized in that the peripheral venous blood of patients with autoimmune thyroiditis determine the absolute content of T-lymphocytes with the transferrin receptor (CD71+) and the concentration of interleukin-2 (IL-2); when the content of lymphocytes CD71+ more than 0.3×109CL/l and the concentration of IL-2 less than 25 PG/ml predicts favorable course of the disease, and when the content of lymphocytes CD71+ less than 0.3×109CL/l and the concentration of IL-2 for more than 25 PG/ml predict an unfavorable course of autoimmune thyroiditis.
SUBSTANCE: patient's peripheral blood is examined for the content of cytotoxic lymphocytes, %: (CD8/CD16), (CP8/GranzymeB), (CP16/GranzymeB), (CDS), (CD16/CD56) and (CD8/HLA DR). It is conmbined with determining cytotoxic activity (CTA), %. It is followed by calculating a RA activity index (AI) by formula: If observing the condition 11.8%<AI≤18.2%, a moderate degree of rheumatoid arthritis activity is diagnosed. If observing the condition AI>18.2%, a high degree of rheumatoid arthritis activity is diagnosed.
EFFECT: using the technique enables high-reliability differential diagnosis of the degree of rheumatoid arthritis activity.
SUBSTANCE: there are involved recording clinical and laboratory manifestations of a CNS injury during the first days of the disease that is followed by the calculation of total diagnostic coefficients related to the detected graduation levels of pathognomonic signs of the disease. Total (+)7 points and more is related to predicting the developing mixed tick-borne encephalitis and borreliosis infection. Total (-)8 points and less shows the developing potential monoinfection of tick-borne encephalitis. If deriving the intermediate values of total diagnostic coefficients when none of said limits is reached, the prognosis is uncertain.
EFFECT: using the method for stating basic tendencies of the developing pathological process at the early stages of the developing disease.
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SUBSTANCE: with using indirect immunofluorescence and monoclonal antibodies, the patients suffering gastric ulcer (GU) are examined for an expression level of CD8, CDDR markers by peripheral blood lymphocytes with the CD8 level exceeding 24.5% and the CDDR level exceeding 15% enabling diagnosing a severe clinical course of GU.
EFFECT: use of the offered diagnostic technique provides high information value in diagnosing the severe clinical course of gastric ulcer, well-timed prediction of the severe clinical course of GU, instant diagnosing, provides well-timed and adequate complex of therapeutic measures.
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SUBSTANCE: method involves the peripheral blood analysis for T-lymphocyte count with receptor expression in relation to CD3+ and CD25+ cells by cell count in 1 mcl of blood before the beginning of treatment and 10 days after. If observing the count increased 7% and more in comparison with the reference, a therapeutic effect in non-psychotic versions of psycho-organic syndrome is predicted to be positive.
EFFECT: use of the method enables more precise prediction of the clinical effectiveness in the given disease that promotes selecting an adequate therapy and reducing length of treatment.
SUBSTANCE: invention describes a method of detecting the decreased phosphatidyl ethanolamine content in red-cell membranes in a pregnant woman suffered aggravated herpes-virus infection in their third trimester of pregnancy with underlying increasing peripheral blood TNFα, characterised by the fact that peripheral blood of the pregnant woman is analysed for the TNFα content to detect phosphatidyl ethanolamine in red-cell membranes by two-dimensional thin-layer chromatography, and a discriminant equation is presented. Df=+1.743×TNFα+(-1.079×phosphatidyl ethanolamine), and if Df is more than a boundary value 78.86, the decreased phosphatidyl ethanolamine content in red-cell membranes is detected.
EFFECT: higher red-cell membrane stability in peripheral blood of the pregnant woman.
SUBSTANCE: invention describes a method for prediction of developing suppurative complications of acute pancreatitis involving laboratory blood analysis wherein the patients are examined for the kinetics of leukocyte chemiluminescence with using an activated suspension of clinical strains Acinetobacter baumannii or Pseudomonas aeruginosa or methicillin-resistant Staphylococcus aureus - MRSA and combining maximisation time - T max more than 1400.0 sec., maximum intensity - 1 max less than 5131.5 standard units, light sum - S less than 144000.0 standard units for the reaction initiated by Acinetobacter baumannii, and combining the values T max>1435.0 sec., I max<4708.0 standard units, S<118000.0 standard units for the reaction initiated by Pseudomonas aeruginosa, or combining the values T max>3520.0 sec., I max<4257.0 standard units, S<169000.0 standard units for the reaction initiated by MRSA enables predicting developing suppurative complications.
EFFECT: higher sensitivity and reduced time of prediction of suppurative complications of acute pancreatitis ensured by enabled assessment of the specific properties of patient's phagocytic response to etiologically significant activators.
1 ex, 4 tbl
SUBSTANCE: invention describes a method of assessment of the effect of arginine dehydrogenase activity on the blood histamine level in pregnant women suffered aggravated herpes viral infection with herpes antibody titre 1:12800 in the third trimester of gestation wherein syncytiotrophoblast of placental villi are examined with a histochemical reaction; section regions are photographed, entered in the computer and cytophotometried; observing increasing arginine dehydrogenase activity up to 48.7±2.9 standard units enables stating increasing the peripheral blood histamine level of the pregnant woman up to 1.44±0.09 mcmole/l that exacerbates foetoplacental deficiency in pregnant women.
EFFECT: method allows assessing the effect of arginine dehydrogenase activity on the blood histamine level of the pregnant women suffered aggravated herpes viral infection in the third trimester of gestation.
SUBSTANCE: pre-therapy blood plasma is analysed for immunoglobulin E, adrenocorticotrophin and endorphin, the index SCORAD dynamics is determined by formula: Dsk=1.34147-0.000820741*IgE-0.00382642*ACTP+0.0209401*endorphin wherein IgE is the level of acute blood plasma immunoglobulin E mIU/ml; ACTP is the level of acute blood plasma adrenocorticotrophin ng/l. If observing the Dsk value exceeding 1.18, a prognosis of improving clinical dynamics of recovery is favourable; the conventional therapy is considered to be sufficient. The Dsk value being equal to 1.18 or less, the improving clinical dynamics of recovery is unfavourable, the intensified therapeutic scheme for atopic dermatitis is considered to be preferential.
EFFECT: higher prediction accuracy.
SUBSTANCE: what is presented is a method for prediction of a character of an uterine involvement in myomatous nodes in a female patient suffering hysteromyoma involving DNA recovery from peripheral venous blood, analysis of +36 A/G locus genotypes of type 1 tumour necrosis factor receptor TKFR1 and analysis of myomatous nodes, and also measurement of an uterus. If detecting the genotypes +36 GG and +36 AG TNFR1, a lesser degree of the uterine involvement is predicted. The genotype +36 AA TNFR1 enables to predict a greater degree of the uterine involvement.
EFFECT: using the presented method allows predicting the character of the early uterine involvement in myomatous nodes.
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SUBSTANCE: for the purpose of diagnosing metabolic compensation disturbances and predicting a risk of developing complications in the patients suffering type 2 diabetes mellitus (type 2 DM), peripheral venous blood is examined for a percentage of apoptotic lymphocytes An+/Pl-. The lymphocyte An+/Pl- value below 3.5 % enables diagnosing a condition of carbohydrate metabolism compensation and predicting a favourable clinical course of the disease for the following year. The lymphocyte An/PI- content within the range 3.5 to 6% provides diagnosing a condition of subcompensation and a moderate risk of developing complications. The An+/Pl- value exceeding 6 % described a condition of decompensation and a high risk of developing chronic complications of type 2 diabetes mellitus.
EFFECT: higher accuracy of prediction and reduction of the risk of developing complications in diabetic patients.
FIELD: medicine, ophthalmology.
SUBSTANCE: in lacrimal liquid one should detect the content of interleukin 8 (IL-8) and that of interleukin 1 beta (IL-1β) to calculate prognostic coefficient (PC) due to dividing the first value by the second one by the following formula: At PC value being below 10.0 one should predict favorable disease flow, and at PC value being above 10.0 - unfavorable flow.
EFFECT: higher accuracy of prediction.
FIELD: medicine, medicinal microbiology.
SUBSTANCE: method involves growing microorganism culture to be studied in solid nutrient medium followed by preparing microbial suspension and its incubation in the presence of lactoferrin. Control sample is prepared in parallel series. Control and experimental samples are incubated, supernatant is removed from bacterial cells and lactoferrin concentration is determined in supernatant of experimental and control sample by immunoenzyme analysis. Then anti-lactoferrin activity is calculated by difference of concentrations of residual lactoferrin in experimental and control samples. This method provides enhancing the sensitivity and precision in carrying out the quantitative evaluation of anti-lactoferrin activity in broad spectrum of microorganisms that is urgent in diagnosis and prognosis of diseases with bacterial etiology. Invention can be used in determination of persistent indices of microorganisms for assay of their etiological significance in pathological processes.
EFFECT: improved assay method.
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FIELD: medicine, biology.
SUBSTANCE: invention relates to nutrient medium used for accumulation of cells for the following cytological and/or immunocytochemical analysis carrying out. Invention relates to medium containing salts NaCl, KCl, anhydrous CaCl2, MgSO4 x 6 H2O, MgCl2 x 6 H2O, Na2HPO4 x 2 H2O, KHPO4, NaHCO3, and also glucose and Henx's solution, 10% albumin solution and polyglucin taken in the ratio 1:1:1. Invention provides enhancing the preservation of cells.
EFFECT: improved an valuable properties of nutrient medium.
FIELD: medicine, cardiology.
SUBSTANCE: in peripheral blood one should detect the level of CD95(+) and CD16(+) neutrophilic granulocytes and at combination of increased level of CD95(+) neutrophilic granulocytes by 4 times and more and CD16(+) neutrophilic granulocytes by 0.6 times against the norm with ECG signs of myocardial infarction one should predict lethal result of large-focal myocardial infarction.
EFFECT: higher accuracy of prediction.
FIELD: medicine, parasitology.
SUBSTANCE: one should carry out immunoenzymatic assay to detect diagnostic optic density and that of labeled immune complex in a plot's hole with tested serum measured in conventional units at wave length being 492 nm. One should calculate coefficient of antibodies concentration measured in conventional units by the following formula: CAC = (Odtsh - Odd) x 100, where CAC - coefficient of antibodies concentration, Odtsh - optic density of the hole with tested serum, Odd - diagnostic value of optic density, 100 - coefficient of serumal dilution. By CAC value one should detect the titer of antibodies to Lamblia intestinalis antigens to interpret results of the trial. The method enables to study the dynamics of disease flow.
EFFECT: higher efficiency and accuracy of diagnostics.
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SUBSTANCE: the present innovation deals with studying and treating diseases of inflammatory, autoimmune and degenerative genesis. One should perform sampling of heparinized blood followed by its sedimentation to obtain blood plasma with leukocytes and centrifuging to isolate the latter which are washed against erythrocytic and serumal admixtures, and, also, it deals with calculating the number of cells in samples out of leukocytic suspension after incubation (B) for 1.5 h at 37 C in holes of plastic microplotting board, out of leukocytic suspension one should additionally prepare two samples, one should be applied to calculate total number of leukocytes before incubation (A), the second sample undergoes incubation at the same mode at addition of autoserum to calculate the number of cells remained after incubation (C). One should state upon adhesive properties of leukocytes by the index of spontaneous adhesion (D), where D=(A-B)/B.100%, and effect for enhanced cellular adhesion under the impact of autoserum should be detected by the value of K=(B-C)/C.100% at K ≥ 30%, where B - C - the number of cells undergone additional adhesion after addition of autoserum. The present innovation widens functional possibilities of the suggested method due to obtaining additional values depicting adhesive properties of blood leukocytes.
EFFECT: higher accuracy of detection.
FIELD: medicine, immunology.
SUBSTANCE: one should carry out reaction of blast-transformation, detect proliferation of T-lymphocytes activated with antibodies to CD3 in the presence of interleukin-7 (ACT IL-7) and in the presence of interleukin-7 and dexametazone (ACT IL-7 D), calculate the index for dexametazone action as the ratio of ACT IL-7 to ACT IL-7 D, moreover, the value of dexametazone action index being above 1.2 indicates increased production of cytokins that suppress T-lymphocytes in neonatals. The method enables to detect functional defect of immune system that characterizes neonatal period.
EFFECT: higher efficiency of detection.
SUBSTANCE: method involves measuring forced exhalation volume per 1 s (FEV1) in l, full right ventricle evacuation time (RVE) in ms and angiotensin II value (AII) in ng/l. Discriminant relationship is built as D=0.504·RVE+3.038·FEV1 - 2.0·AII. D being less than 83.88, pulmonary hypertension occurrence is predicted within 1 year. D being equal to or greater than 83.88, no pulmonary hypertension is predicted to occur.
EFFECT: enhanced accuracy of prediction.
FIELD: medicine, medicinal immunology.
SUBSTANCE: method involves determination of heterophilic antibodies in human serum blood by the Paul-Bunnel's method relatively the level of circulating immune complexes, complement-activating properties of heterophilic antibodies by incubation of standardized ram erythrocytes with 0.8% serum for 30 ± 5 min and the following measurement of the erythrocytes lysis degree. The measurement of the effector function coefficient of heterophilic antibodies is carried out by the complement system Keff.f.h.a.-c.s. by the formula: Keff.f.h.a.-c.s. = Y/Tg.a. wherein Y means a lysis degree, %; Tg.a. means a reverse titer of heterophilic antibodies to ram erythrocytes. The damage assay is carried out by comparison of the immune status with the relative level of circulating immune complexes in serum. Method provides detection of preclinic from of immunodeficiency and autoimmune diseases that opens the possibility for their prophylaxis at most early stages of development. Invention can be used for assay of damage in the immune status in human serum blood.
EFFECT: improved method for assay.
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SUBSTANCE: method involves concurrently examining anti-inflammatory IL-4 level in blood serum and lacrimal fluid. The value being within the limits of 60-70 pg/l in blood serum and 5-15 pg/l in lacrimal fluid, disease prognosis is considered to be unfavorable. The IL-4 concentration being within the limits of 90-100 pg/l in blood serum and 20-30 pg/l in lacrimal fluid, disease prognosis is considered to be favorable.
EFFECT: high accuracy of diagnosis.