ABSTRACT
OBJECTIVE: Studying the effectiveness of the combined use of intraosseous blockades (IOB) and intramuscular local injection therapy using bottled forms of HYALREPAIR-02 Chondroreparant 02/10 in patients with lumbar dorsopathy and recurrent back pain syndrome. MATERIAL AND METHODS: 30 patients (16 men and 14 women) with chronic recurrent lumbar dorsopathy with pain and myofascial syndromes were examined. Group 1 included 17 patients, whose treatment included the use of IOB and intramuscular local injection therapy using bottled forms of HYALREPAIR-02 Chondroreparant 02/10. Group 2 included 13 patients who received only IOB. To assess the pain syndrome, a visual analogue pain scale (VAS), a Russian version of the McGill Pain Questionnaire (MPBI), and a body diagram questionnaire were used. RESULTS: After the course of treatment, in both groups there was a decrease in the severity of pain according to VAS, RMBO, and a decrease in the area of pain distribution according to the Body Scheme questionnaire (p<0.05). The values on the RMBO questionnaire in both groups also decreased by more than 2 times compared to the initial value. A statistically more significant regression of pain syndrome and a decrease in disability were noted in group 1 (p<0.05). 2 months after the end of the course of treatment, the pain syndrome did not recur in any patient. Within 3 to 6 months, back pain recurred in 4 (23.5%) patients of the 1st group and in 6 (46.2%) patients of the 2nd group. A total of 90 IOB procedures were performed; no complications or side effects were noted. CONCLUSION: The combination of IOB and intramuscular local injection therapy using the vial form of HYALREPAIR-02 Chondroreparant in the treatment of patients with degenerative diseases of the spine and back pain is an effective and safe method of therapy. Such treatment strategy allowed to relieve back pain more effectively and improved long-term clinical outcome.
Subject(s)
Low Back Pain , Spinal Diseases , Male , Humans , Female , Low Back Pain/drug therapy , Injections, Intramuscular , Lumbosacral Region , SpineABSTRACT
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.
Subject(s)
Leukemia, Lymphocytic, Chronic, B-Cell , HLA-DR Antigens , Humans , Immunity, Cellular , Immunophenotyping , Killer Cells, Natural , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapyABSTRACT
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.
Subject(s)
Complement Membrane Attack Complex/metabolism , Erythrocyte Membrane/metabolism , Flow Cytometry/methods , Hemoglobinuria, Paroxysmal/blood , Hemoglobinuria, Paroxysmal/diagnosis , Membrane Proteins/blood , Chromosomes, Human, X , Complement Activation/genetics , Complement Membrane Attack Complex/genetics , Diagnosis, Differential , Erythrocyte Membrane/genetics , Female , GPI-Linked Proteins/blood , GPI-Linked Proteins/genetics , Hemoglobinuria, Paroxysmal/genetics , Humans , Male , Membrane Proteins/genetics , Mutation , Reticulocyte CountABSTRACT
In the past decade, a notable advance has been made in the understanding of the pathogenesis of NK/T-cell lymphomas; however, their diagnosis remains difficult because of their rarity and clinical and morphological variabilities. The paper generalizes the ten-year experience of the Hematology Research Center, Ministry of Health of Russia, in diagnosing and treating hepatosplenic T-cell lymphoma (HSTL), considers the problems of differential diagnosis with other hematological diseases occurring with similar clinical and laboratory symptoms, and lays down current approaches to the diagnosis and treatment of this condition. A clinician's view of the problem of diagnosis and treatment of this disease is given. HSTL is shown to be a heterogeneous group of diseases differing in a T-cell receptor chain gene rearrangement, the clinical course of the disease, and overall survival (OS). According to our data, 3-year OS was 12%; the median survival was 26 months. Two-year OS for γδ and αß HSTL was equal to 25 and 70%, respectively. The difference in OS for the variants of HSTL failed to reach statistical significance (because the sample might be insufficient).
Subject(s)
Lymphoma, T-Cell/diagnosis , Humans , Lymphoma, T-Cell/therapy , Prognosis , RussiaABSTRACT
The implementation of principles of highly sensitive flow cytometry into diagnostic of paroxysmal nocturnal hemoglobinuria increased rate of detection of paroxysmal nocturnal hemoglobinuria clone in patients with aplastic anemia already at early stages of diagnosis establishment (up to 79%). However, detection of paroxysmal nocturnal hemoglobinuria clone attracts interest not only from point of view of progression in % of patients with aplastic anemia). The occurrence of paroxysmal nocturnal hemoglobinuria clone in patients with aplastic anemia can be accompanied by hidden disorders of haemopoesis with increasing risk in conditions of proliferative stress. Hence, it is necessary to monitor the given clone during all period of observation. The study is a prospective investigation analyzing dynamics of paroxysmal nocturnal hemoglobinuria clone in process of immune suppressive therapy applied to 44 patients with aplastic anemia. The mentioned clone was initially detected in 59.6% of patients. The median of observation amounted to 27 (9-48) months. Depending on size of granulocytic paroxysmal nocturnal hemoglobinuria clone patients were allocated in four conditional groups: group I - from 0.01% to 0.99% (n=11); group II - from 1% to 9.99% (n=8); group III - from10% to 49.9% (n=4); group IV - from 50% and more (n=5). In the course of study the differently directed dynamics of paroxysmal nocturnal hemoglobinuria clone was revealed. In 3 out of 11 patients from group I median of paroxysmal nocturnal hemoglobinuria clone increased from minor values (less than 1%) to 3.55%; at that in one patient occurred total elimination of paroxysmal nocturnal hemoglobinuria clone to 12th month of observation. The noticeable unidirectional dynamics was established in patients of group III: already to 3d month of observation, simultaneously with becoming of remission, median of size of paroxysmal nocturnal hemoglobinuria clone in group diminished from 22.9% (18.39%-24.77%) to 5.6% (1.5%-6.7%). Among patients of groups II and IV paroxysmal nocturnal hemoglobinuria clone remained stable. The development of hemolytic form of paroxysmal nocturnal hemoglobinuria was observed in all patients of group IV i.e. in 18% of patients with aplastic anemia with primarily detected paroxysmal nocturnal hemoglobinuria clone. In the process of observation, in 37% of patients with aplastic anemia without primarily detected paroxysmal nocturnal hemoglobinuria clone its occurrence and persistence (median - 0.34% (0.1%-6.2%)) was noticed. According to the results of study, alteration of sizes of paroxysmal nocturnal hemoglobinuria clone or its occurrence develop in case of response to ISP and, most probably, depend on advantage of growth in the process of repair of normal (GPI positive) or clonal (GPI negative) hemopoiesis. To acquire more reliable conclusions will be possible through development of techniques of molecular diagnostic simultaneously with dynamic observation of course of disease in the given patients.
Subject(s)
Anemia, Aplastic/blood , Flow Cytometry , Hemoglobinuria, Paroxysmal/blood , Membrane Proteins/genetics , Anemia, Aplastic/complications , Anemia, Aplastic/genetics , Erythrocytes/metabolism , Erythrocytes/pathology , Female , Glycosylphosphatidylinositols/biosynthesis , Granulocytes/metabolism , Granulocytes/pathology , Hemoglobinuria, Paroxysmal/complications , Hemoglobinuria, Paroxysmal/genetics , Humans , Male , Monocytes/metabolism , Monocytes/pathologyABSTRACT
The paroxysmal nocturnal hemoglobinuria is a rare clonal disease characterized by somatic mutation of gene PIG-A at the level of stem hematopoietic cell. This process results in disorder of synthesis of glycosil phosphatidyl innozitol (GPI) anchor fixing numerous molecules on membrane of blood cells which protect blood cells from impact of complement. The international society of clinical cytometry (2010) proposed the guidelines of detection of clone of paroxysmal nocturnal hemoglobinuria among erythrocytes, granulocytes and monocytes. The original technique is proposed to evaluate the clone of paroxysmal nocturnal hemoglobinuria in reticulocyte population of blood using method of flow cytofluorometry. The sampling of 160 samples of blood of patients with clinical symptoms of paroxysmal nocturnal hemoglobinuria and anemia was analyzed. Two modes of gatedrawing were applied--using monoclonal antibodies to CD71 (receptor to transferrin) and reagent BD ReticCount. The high correlation was established between size of reticulocytic clone of paroxysmal nocturnal hemoglobinuria evaluated by CD71 and size of granulocytic and monocytic clone of paroxysmal nocturnal hemoglobinuria. The developed panel (CD71/CD235a/CD59) can be applied for screening and monitoring of paroxysmal nocturnal hemoglobinuria.
Subject(s)
Antigens, CD/blood , Flow Cytometry/methods , Hemoglobinuria, Paroxysmal/blood , Receptors, Transferrin/blood , Reticulocytes/metabolism , Adolescent , Adult , Aged , Antibodies, Monoclonal/chemistry , CD59 Antigens/blood , Female , Hemoglobinuria, Paroxysmal/diagnosis , Humans , Male , Middle AgedABSTRACT
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.
Subject(s)
Drug-Related Side Effects and Adverse Reactions/blood , Flow Cytometry , Leukemia, Lymphocytic, Chronic, B-Cell/blood , Neoplasm, Residual/blood , Adult , Aged , Aged, 80 and over , Antigens, CD/blood , Drug-Related Side Effects and Adverse Reactions/pathology , Female , Humans , Leukemia, Lymphocytic, Chronic, B-Cell/complications , Leukemia, Lymphocytic, Chronic, B-Cell/drug therapy , Male , Middle Aged , Neoplasm, Residual/chemically induced , Neoplasm, Residual/pathology , Predictive Value of Tests , PrognosisABSTRACT
The flow cytometry becomes a more and more largely applied technique. However, the sufficient novelty of technique has no worked-out standards of diagnostic of many diseases. The lacking of external control of quality promotes development of large variety of approaches to diagnostic of diseases and impossibility to compare the study results from different laboratories. The paroxysmal night hemoglobinuria is an acquired clonal disease characterized by proliferation of stem cells with partial or total loss of expression of glykosylphosphosphatidyl inositol anchor needed to conjugate a number of surface proteins. The flow cytometry is a basic technique of detection and monitoring of clone of paroxysmal night hemoglobinuria. The article presents the results of paroxysmal night hemoglobinuria testing of 8 patients in 6 independent laboratories using flow cytometry by standard protocol recommended by the International society of clinical cytometrists (ICCS).
Subject(s)
Flow Cytometry/standards , Hemoglobinuria, Paroxysmal/diagnosis , Adult , Female , Flow Cytometry/methods , Hemoglobinuria, Paroxysmal/blood , Humans , Male , Middle AgedABSTRACT
The techniques of flow cytofluorometry and immune cytochemistry combined with routine cytological analysis were applied to 81 patients with non-Hodgkin's lymphomas to diagnose the malignant lymphomas and to determine their immune phenotypes. The accuracy of proposed techniques combination to diagnose the malignant lymphoma consists 98% and to determine the immune phenotype - 90%. The presented combination of routine cytological analysis combined with techniques of flow cytofluorometry and immune cytochemistry is the express-technique to diagnose lymphoma and its immune phenotype the day of patient visit to the doctor. This approach significantly increases the role of cytological method in diagnostics of lymphomas. The technique makes it possible to analyze simultaneously huge quantity number of lymph nodes, including deep-seated and not only the superficial lymph nodes.
Subject(s)
Flow Cytometry/methods , Lymphoma, Non-Hodgkin/blood , Lymphoma, Non-Hodgkin/diagnosis , Humans , Lymphoma, Non-Hodgkin/pathology , Sensitivity and SpecificityABSTRACT
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.
Subject(s)
Flow Cytometry/methods , Gene Expression Regulation, Leukemic , Leukemia, Lymphocytic, Chronic, B-Cell/blood , Neoplasm Proteins/biosynthesis , ZAP-70 Protein-Tyrosine Kinase/biosynthesis , Adult , Aged , Aged, 80 and over , Female , Humans , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Heavy Chains/metabolism , Immunoglobulin Variable Region/genetics , Immunoglobulin Variable Region/metabolism , Leukemia, Lymphocytic, Chronic, B-Cell/diagnosis , Leukemia, Lymphocytic, Chronic, B-Cell/genetics , Leukemia, Lymphocytic, Chronic, B-Cell/therapy , Male , Middle Aged , MutationABSTRACT
The content of CD34/CD45dim-positive cells in peripheral blood of children with congenital and hereditary diseases of hepatobiliary system is studied. The analysis of relationship between numbers of studied cells and level of C-reactive protein, sCD40L, sCD30 and laboratory parameters specific to liver functions is applied The number of CD34-positive hemopoietic hematoblasts in children with hepatocirrhosis correlated with the level of C-reactive protein, albumin, hemoglobin concentration and quantity of blood erythrocytes. No relationship was established with the levels of sDC40L and sDC30. The number ofstudied cells in children with liver diseases was higher than in healthy adult donors.
Subject(s)
Biliary Tract/pathology , Hematopoietic System , Liver Cirrhosis , Liver Transplantation/methods , Adult , Antigens, CD34/adverse effects , Antigens, CD34/blood , C-Reactive Protein/analysis , Child, Preschool , Erythrocyte Count , Female , Hematopoietic Stem Cells/cytology , Hematopoietic System/pathology , Humans , Infant , Leukocyte Common Antigens/blood , Leukocyte Count , Liver Cirrhosis/blood , Liver Cirrhosis/pathology , MaleABSTRACT
The article considers the main positions concerning the clinical laboratory diagnostics as an independent clinical specialty and the principles of professional training and improvement of specialists. The basic issues complicating the training and improvement of personnel to be kept in line with actual needs of laboratory service of public health system are discussed. Among them are the availability of laboratory academic sub disciplines demanding a profound special theoretical education and technical skills; the need to account in the process of professional training the variety of forms, sizes and types of laboratory structures in different medical institutions; the need of special training programs for numerous specialists with non-medical basic education. The combination of the present system of postgraduate training of specialists on chairs of state educational organizations with initiative involvement of specialists in various public forms of permanent professional improvement (professional scientific societies meetings, research conferences, internet seminars, etc.) is supported Along with a positive appraisal of the existing system of training in the state educational institutions and corresponding regulation documents, a critique is expressed regarding certain actual documents which improperly limit the administrative functions of physicians of clinical laboratory diagnostics and complicate training of bacteriologists for clinical laboratories.
Subject(s)
Diagnostic Services , Education, Medical, Continuing , Medical Laboratory Personnel/education , Diagnostic Services/legislation & jurisprudence , Diagnostic Services/standards , Humans , Medical Laboratory Personnel/classification , Medical Laboratory Personnel/legislation & jurisprudence , Medical Laboratory Personnel/standards , Physicians/standardsABSTRACT
The new effective protocols of treatment of chronic B-cell lymphatic leukemia, including purine analogs and monoclonal antibodies, provide robust remissions under this disease. Accordingly, the requirements to remission quality assessment are changed too. In particular the assessment of minimal residual disease is obligatory. To assess minimal residual disease in terms of quantity in case of chronic B-cell lymphatic leukemia the technique of polymerase chain reaction was applied in real time with patient-specific primers from the area of V-D-J combinations of genes of heavy chain of immunoglobulin. The study included samples from 60 patients suffering of chronic B-cell lymphatic leukemia. In 15 of them (25%), it was impossible to apply neither the sequence analysis of genes of heavy chain of immunoglobulin nor the fitting of patient-specific primer. The results of quantitative determination of minimal residual disease were obtained in 45 patients (55 tests). The minimal residual disease was detected in 30 of 55 samples (54.5%) and was not detected in 25 of 55 samples (45.5%). At the same time, the quantitative determination of minimal residual disease was implemented in regard to the initial level of neoplastic cells. The method sensitivity qualified by serial dilutions, consisted 10(-5) or 1 neoplastic cell to 100 000 normal cells. The comparative analysis was applied to the results of determination of minimal residual disease using two methods -polymerase chain reaction in real time using patient-specified primers and four-color flow cytofluometry. The determination of minimal residual disease with both methods was implemented in 37 patients (45 tests). The results of both methods matched in 93.3% (42 tests out of 45) with maximal disparity of one degree. Then Spearman factor consisted 0.87 (p < 0.0001). In 3 out of 45 tests (6.7%) neoplastic cells were detected with only one method. In the first case, it was the method of four-color flow cytofluometry and in other two cases it was polymerase chain reaction in real time. Therefore, the detection of minimal residual disease under chronic B-cell lymphatic leukemia using the method of polymerase chain reaction in real time is rather sensitive and specific and correlates with the results received with the method of four-color flow cytofluometry. The results are the same in the case of using anti-CD20 monoclonal antibodies under treatment.