Educational video for training specialists in clinical laboratory diagnostics.

The adaptation of educational programs under restrictions during the SARS-CoV-2 coronavirus pandemic and after it is based not only on the widespread use of video lectures, but also on the introduction of a number of new educational technologies. Hybrid learning will become the cornerstone of future educational technologies in clinical laboratory diagnostics and will contribute to the creation of virtual practical classes with examples of analysis of laboratory testing results based on case histories (Case Technologies). The key aspect in the implementation of video cases into the educational process is the development of video materials. The use of educational video cases developed by Vector-Best in the process of training specialists in clinical laboratory diagnostics during the cycles «Retraining¼ and «General Improvement¼ at the Department of Clinical Laboratory Diagnostics of the Saratov State Medical University named after V. I. Razumovsky of the Ministry of Health of Russia was a convenient format and received positive assessment of cadets. Currently, the «IVD gallery¼ section has appeared on the FLM website and the placement of an additional educational resource - a library of educational video cases.

Specific features of T- and NK-cellular immunity in chronic lymphocytic leukemia.

Profound immunological dysfunction is the key factor determining the development of infectious complications in chronic lymphocytic leukemia (CLL). The aim of this work is to assess the features of the subpopulation composition of T-lymphocytes (T-helpers (Th), cytotoxic T-lymphocytes (Tcyt), T regulatory cells (Treg), T-NK cells, naive Th, Th-memory, activated T-lymphocytes, TCRγδ cells) and NK cells in peripheral blood of patients with newly diagnosed chronic lymphocytic leukemia (CLL) and receiving ibrutinib therapy. Hematological and immunophenotypic studies have been performed in 30 patients with previously untreated CLL, 122 patients on ibrutinib therapy and 20 healthy donors. The subpopulation composition of T-lymphocytes (Th, Tcyt, Treg, T-NK, naive T-helpers, memory T-helpers, TCRγδ cells, activated T-lymphocytes) and NK cells has been assessed on flow cytometer (FACSCanto II (BD)) using the following panel of monoclonal antibodies: CD45, CD19, CD3, CD4, CD5, CD8, TCRγδ, CD127, CD16, CD56, CD57 CD45RA, CD45R0, HLA-DR, CD25. Compared to controls all CLL samples were found to have higher the absolute number of T-lymphocytes, NK cells and their subpopulations, T-helpers (especially of memory T-cells), cytotoxic T-cells, regulatory T-cells, TCRγδ T-cells, activated T-lymphocytes, increased cytotoxic potential of NK cells in previously untreated CLL patients. Patients who received ibrutinib therapy have registered a positive trend towards recovery of the subpopulation composition of T-lymphocytes and NK-cells. CLL patients have been found to have quantitative and functional changes in the subpopulations of T-lymphocytes and NK cells, indicating dysregulation of the immune response, and a high risk of developing infections. Monitoring of immunological parameters for ibrutinib therapy make possible to estimate impact of ibrutinib on the adaptive anti-CLL immune response.

[The diagnostic screening of acute leukemia using new technologies of automated blood analysis.]

The leukemia is a neoplastic clonal disease of hematopoietic system with primary affection of bone marrow. The modern technologies of automated blood analysis permit to implement a quick primary screening of pathological samples of blood suspicious for presence of blast cells. The application of various analysis techniques (optical, cytochemical, immune phenotypical) with the purpose of detecting blast cells demonstrates their different distribution at the graphics.

[Glycemic variability and oxidative stress in patients with type 2 diabetes mellitus during combined glucose-lowering therapy]. / Variabel'nost' urovnia gliukozy v krovi i okislitel'nyi stress u bol'nykh sakharnym diabetom 2-go tipa na fone kombinirovannoi sakharosnizhaiushchei terapii.

AIM: To evaluate the impact of intensified glucose-lowering therapy on carbohydrate metabolic indicator, such as glycated hemoglobin, fasting blood glucose level (BGL) (FBGL), postprandial BGL (PBGL), and glycemic variability (GV) in patients with type 2 diabetes mellitus (T2DM) during metformin monotherapy before and 3 months after therapy intensification. SUBJECTS AND METHODS: The investigation enrolled 51 patients with T2DM treated with metformin 1000 mg twice daily, who failed to achieve satisfactory glycemic control. During randomization, the treatment was intensified by addition of sitagliptin 100 mg/day in Group 1 (n=25) or gliclazide MB 60 mg/day in Group 2 (n=26). Before and 3 months after the treatment, carbohydrate metabolic indicators were investigated, 24-hour BGL monitoring (continuous glucose monitoring system (GMS)) was performed, and the body's antioxidant status was examined by determining the total antioxidant capacity of blood plasma (overall sound pressure levels (OASPL)). RESULTS: During 3-month treatment, Group 1 had a significantly reduced FBGL compared to that before the therapy; in Group 2 this index did not change significantly. Both study groups showed a significant decrease in PBGL and glycated hemoglobin (HbA1c). The mean amplitude of glycemic excursion (MAGE) was significantly decreased in the sitagliptin intensification group. In both groups, the standard deviation (SD) reduced significantly by 26% in Group 1 and by 38% in Group 2. Both groups also displayed a significant increase in blood OASPL (p<0.05). CONCLUSION: The addition of sitagliptin significantly affected the change in the indicators of both the standard carbohydrate metabolism (FBGL, PBGL, and HbA1c) and GV (MAGE, SD), whereas that of gliclazide MV altered some studied parameters. OASPL significantly increased in both groups.

[THE METHODICAL APPROACHES TO DIAGNOSTIC OF NIGHT PAROXYSMAL HEMOGLOBINURIA].

The article presents diagnostic of night paroxysmal hemoglobinuria. The night paroxysmal hemoglobinuria is an orphan disease characterized by absence of GPI-anchor on blood cells as a result of mutation of PIG-A gene on the short arm of X-chromosome. The particular proteins bounded with GPI-anchor implement function of defense from activation of components of complement and development of membrane-attacking complex. The erythrocytes exposed to destruction in bloodstream are among the most impacted. Therefore, one of the main signs of night paroxysmal hemoglobinuria is complement-depending intravascular hemolysis which indicators for a long time played a key role in diagnostic of night paroxysmal hemoglobinuria. The actual technique of diagnostic of night paroxysmal hemoglobinuria is flow cytometry. The analysis of night paroxysmal hemoglobinuria clone is recommended to patients with hemolysis of unclear genesis, thrombosis of cerebral and abdominal veins, thrombocytopenia and macrocytosis and also patients with AA, myelodysplastic syndrome, myelofibrosis. The international protocol recommended by the International Society of Clinical Cytometry (2010) is implemented to diagnose night paroxysmal hemoglobinuria. The original technique of evaluation of reticulocytes was developed with purpose to detect night paroxysmal hemoglobinuria clone. The high correlation was substantiated between size of night paroxysmal hemoglobinuria clone measured among reticulocytes according to proposed mode and night paroxysmal hemoglobinuria clone measured among granulocytes and monocytes detected according international standardized approach.

[THE CONTENT OF ELECTROLYTES IN DIFFERENT BIOLOGICAL MEDIUMS UNDER ACUTE CORONARY SYNDROME].

The sampling of study included 172 patients with ischemic heart disease: 146 with acute coronary syndrome, including myocardial infarction (88 patients) and unstable stenocardia (58 patients); 26 patients with stable stenocardia functional class II-III. At the 1-3 day of hospitalization blood was taken of cubital vein. The mixed unstimulated saliva was selected. In both of them conte of calcium, sodium and potassium was tested (mmol/l). Under acute coronary syndrome, in blood content of calcium was 2. sodium--139.6 and potassium--4.5 i.e. the content was lower than in case of stable stenocardia (2.4; 139.8; 4.7 correspondingl In saliva under acute coronary syndrome higher content of calcium (1.05) and potassium (34.66) and lower content of sodiu (25.42) was established in comparison with stable stenocardia (0.81; 33.7; 28.08 correspondingly). The distribution coefficien (blood/saliva) of calcium, sodium and potassium were higher under myocardium infarction than under unstable stenocardia at uncomplicated course of acute coronary syndrome.

[Electronic textbooks and libraries abroad].

Electronic textbooks and libraries abroad. - The current article presents a brief analysis of the modern state and perspectives of electronic textbooks and libraries abroad. The material is based on the open access foreign internet sources. Using particular examples it has been shown that despite a long history of electronic textbooks and libraries development, the latter remains at the very early stage. The bureaucratic attempts to speed up this process result in costly failures. It is necessary a painstaking and cautious work in the mode of scientific practical experiment to give electronic textbooks and libraries a chance to make worthy contribution into the progress of training, education and culture.

[The certification and accreditation of clinical diagnostic laboratory].

Two approaches are considered concerning evaluation of quality of functioning of clinical diagnostic laboratory--certification and accreditation. The comparison and juxtaposition of these two approaches is made. The practical importance and applicability of requirements of standards of GOSTK ISO 9001-2008 and GOSTR ISO 15189-2009 is demonstrated. The attention is paid to key issues of workability of system of quality management in laboratory and organization of its technical competence at required level. The standard basics of requirements of international and national quality standards are specified. The article can have crucial practical importance for laboratories which implement system of quality management and prepare for accreditation, pay proper attention to standardization of laboratory studies, strive for workability of system of quality management and determine future priorities in area of enhancement of laboratory services.

[The evaluation of prognostic value of biomarkers in patients with chronic cardiac insufficiency].

Nowadays, the laboratory markers of myocardial dysfunction (cerebral natriuretic peptide - NT-proBNP); instability of atherosclerotic plaque (highly sensitive C-reactive protein - hsCRP); damages of cardiac muscle (highly sensitive cardiac tropine I - hs-cTnl) play a key role in diagnostic, course prognostics and verification of risk of unfavorable outcomes in patients with chronic cardiac insufficiency. The article presents the results of study of dynamics of levels of NT-proBNP, hsCRP, hs-cTnl in 71 patients with chronic cardiac insufficiency of II and III functional class (according classification of New-York association of cardiologists - NYHA). The comparison was made concerning analyzed laboratory markers with fraction of output of left ventricle of heart and index of body mass relatively to their prognostic role inpatients with chronic cardiac insufficiency with different clinical conditions and various outcomes of disease. It is demonstrated that level of hs-cTnl is the most valuable in respect to unfavorable prognosis of course of chronic cardiac insufficiency and risk of lethal outcome. The level of NT-proBNP has invert correlation relationship with fraction of output of left ventricle of heart and can be considered as a laboratory indicator of functional condition of myocardium in patients with chronic cardiac insufficiency.

[The acoustic indicator of saliva under stress].

The situation of stress affects various organs and systems that results in development of functional disorders and/or somatic diseases. As a result, different noninvasive, including salivary, techniques of diagnostic of stress conditions are in the process of development. The dynamics of acoustic indicator of saliva is studied during the period of passing the exams. The relationship of indicator with levels of potassium, sodium, glucose and protein of saliva was analyzed. The sampling consisted of 102 students of 5 and 6 academic years of medical university. To detect the acoustic indicator of saliva acoustic analyzer AKBa-01- "BIOM" was applied. The level of potassium and sodium in saliva was detected using method of flame photometry. The level of glucose in saliva was detected by glucose oxydase technique using analyzer "EXAN-G". The protein in saliva was detected by biuretic technique. The correlation between acoustic indicator of saliva and analyzed indicators of saliva was established.

[The dynamics and prognostic value of cerebral natriuretic peptide and C-reactive protein under acute cardiac infarction depending on tactic of treatment].

The sampling consisted of 102 patients with acute cardiac infarction with ST elevation up to 6 hours from the onset of disease. The subjects were divided into three groups: group I with primary stent procedure, group II with deferred stent procedure during 24 hours after effective thrombolytic therapy and group III with isolated effective thrombolytic therapy. The Doppler-echocardiography was applied to all patients. In every group the evaluation was carried out to dynamics NT-proBNP and C-reactive protein. In group III with isolated effective thrombolytic therapy in sub-acute period of cardiac infarction NT-proBNP increased more than twice in comparison with groups of patients with stent procedure. The values of C-reactive protein were lower than in groups I and II. The dynamics of NT-proBNP and C-reactive protein depend on mode of regeneration of coronary blood flow and level of increase of NT-proBNP corresponds the type of diastolic dysfunction of left ventricle of heart.

[The prognostic value of evaluation of minimal residual disease using technique of flow cytofluorometry during application of therapy of chronic lymphatic leukemia].

The achievement of molecular remission is associated with increasing of survival of patients with chronic lymphatic leukemia. The important direction of research is seeking of parameters applicable to forecast of response to therapy. The purpose of the study was evaluating prognostic significance of indicator of minimal residual disease detected by technique of flow cytofluorometry of peripheral blood of patients with chronic lymphatic leukemia during therapy application. The sampling included 112 patients with chronic lymphatic leukemia aged from 43 to 82 years. All patients were given treatment consisted of 6 courses of immune chemotherapy combined with fludarabine with cyclophosphan and rituximab. The samples of peripheral blood were analyzed after 3 courses during therapy and after 6 courses after completion of treatment. The cells were analyzed using 5 and 6 color flow cytometry for the purpose of detection of immune phenotype associated with chronic lymphatic leukemia. The evaluation of minimal residual disease was implemented according international standardized protocol (Rawstron A.C. et al. 2007; 21 (5): 956-64). The minimal residual disease negative status was reached in 87 (78%) patients during evaluation of response after 6th course of treatment. The implementation of indicators of residual disease after 3 courses with fludarabine, cyclophosphan and rituximab permitted to sort out two groups of patients with chronic lymphatic leukemia i.e 67 patients with low (< 0.12%) level of minimal residual disease and 45 patients with high (> 0.12%) level of tumor cells. The rate of molecular remission after completion of treatment. in the given groups consisted 100% and 44% correspondingly. The study demonstrates possibilities of early immune phenotype evaluation of minimal residual disease to forecast differences in response to treatment in patients with chronic lymphatic leukemia that makes it possible to avoid undesirable toxicity of therapy or to choose method of consolidation.

[Dynamics of laboratory markers of endothelial dysfunction during treatment of unstable angina].

This study was aimed to assess the diagnostic value of laboratory markers of endothelial lesions in patients with unstable angina (UA). Plasma levels of CRP, homocysteine, endothelin-1 (ET1), and pregnancy-associated plasma protein (pAPP-A) were measured by an ultrasensitive method in 51 patients. They were followed up for 4 months to evaluate the clinical picture of CHD and dynamics of laboratory parameters. The hospitalized patients with UA had elevated baseline levels of CRP (5.02+-3.35 mg/ml) during the entire study period. PAPP-A and homocysteine levels were significantly decreased (p<0.05 and p<0.0 respectively). Patients with deteriorated clinical picture of CHD had significantly elevated ET1 levels (p<0.05) both on day 1 and 4 months later It is concluded that ET1 levels increase in patients with UA and severe prognostically unfavourable CHD.

[The characteristics of evaluation of expression of ZAP-70 in tumor cells under b-cell chronic lymphatic leukemia using the flow cytofluorometry technique].

The b-cell chronic lymphatic leukemia is the most common among all lymphatic proliferative diseases and is characterized by significant variability of its clinical course. The mutation status of genes of variable region of heavy chains of immunoglobulins (IgVH) is the most reliable prognostic factor forecasting time until beginning of treatment in case of b-cell chronic lymphatic leukemia. However its detection nowadays is inaccessible for routine diagnostics. Among surrogate markers of mutation status the indicator of expression of ZAP-70 by tumor cells estimated using flow cytofluorometry. However, in publications there are different guidelines concerning the technique of mentioned marker. To establish the optimal approach to evaluation of expression of ZAP-70 the peripheral blood samples of 5I patients with b-cell chronic lymphatic leukemia and 10 healthy persons were analyzed. The comparison with the results of detection of mutation status of IgVH-genes revealed the advantage of applying the technique of calculation of MFI ratio during interpretation of data of expression of ZAP-70 obtained with flow cytofluorometry. In this framework, the indicator of expression of ZAP-70 can be applied in assessing the course of disease and time until the beginning of treatment of b-cell chronic lymphatic leukemia.

[The reference values of indicators of total blood analysis of adult working population].

The application in laboratory diagnostics of the hematologic analyzers of different class functioning on the basis of various methods of calculation and identification of blood cells condition the need of establishing the reference intervals for the blood picture parameters. In the documents GOSTK 53002-2008, S28-A3 (CLSI) of 2008 the regulations are proposed to establish the reference intervals with detailed description of data statistical analysis. The processing of blood pictures of 1453 healthy patients the reference intervals were established for the indicators of clinical blood analysis. The results saved in the laboratory data base and obtained using analyzer Sysmex XT-2000i (Sysmex Corporation, Japan) were analyzed.

[Effect of body mass reduction on the level of adiponectin in patients with obesity and type 2 diabetes mellitus].

Patients with obesity and type 2 diabetes mellitus are known to have hypoadiponectinemia partially compensated by switching to low-caloric diet. Reduction of the body mass results in the proportional increase of the adiponectin level and decreased severity of insulin resistance.