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Ex vivo proliferation and differentiation of Philadelphia chromosome-positive (Ph+) human myeloid cells (Ph+ cells) from chronic myeloid leukemia (CML) proceed via alternation stages of cell proliferation and neutrophil maturation. To regulate them, apoptosis is alternately blocked or induced with the help of neutrophils and expression of bcr/abl, bax, and bcl2. The regulation of apoptosis in main types of Ph+ cells depends on the alternation of (1) Ph+ cell proliferation and (2) neutrophil maturation and may follow two pathways. One consists in alternating blockages and inductions of apoptosis with initial maturation and subsequent proliferation under alternation stages as (2)-(1)-(2) and has not been described as yet. Neutrophil accumulation blocks apoptosis. As neutrophils are depleted, apoptosis is induced again. Its block accelerates proliferation with a new accumulation of neutrophils, which is followed by regular neutrophil death and a new induction of apoptosis. The way optimizes the proliferation efficiency (P/D index) with a regular alternation of maturation and proliferation, allowing the cycle of proliferation and differentiation to be completed. In another way, the alternation starts with proliferation as (1)-(2)-(1) at a lower neutrophil content) and leads to resistant decrease of the maximal apoptosis level by a factor of 3–8 as compared with (2)-(1)-(2) alternation. A stable block of apoptosis is observed in cells with prolonged stages of proliferation and maturation, leading to an accumulation of blasts and myelocytes with elevated bcr/abl expression and expression of bcl2 > bax. A stable block of apoptosis is associated with CML progression and in Ph+ cell lines. Cells follow the first pathway of the apoptotic regulation in chronic-phase CML. Ex vivo cultivation of Ph+ cells from individual CML patients was assumed to provide for a more exact diagnosis of the CML phase and optimizing the treatment.

Human myeloid cells with Ph chromosome (Ph+ cells) from chronic myeloid leukemia (CML) in the course of proliferation and differentiation ex vivo are regulated under alternation of cell proliferation and neutrophil maturation stages by consecutive blocking and inducing apoptosis with of neutrophils participation as well bcr/abl, bax and bcl2 genes expression. Apoptosis regulation of three main Ph+ cells types from CML patients depends on alternation sequences of proliferation (1) and maturation (2) cell stages and realized by two ways. The first one is performed by consecutive blocking and inducing apoptosis under 2/1/2 stage alternation. The way is not described early. Neutrophils accumulation correlates with apoprosis blocking. Apoptosis level enhances under neutrophils exhausted. Apoptosis blockage allows cells to proliferate and, thus, to form new portion of neutrophils with consecutive regular their death as well a consequent alternation of apoptosis blocking and inducing. This way regulates proliferation efficiency indexes P/D that reflect Ph+ cells proliferating potential and performs cycle completion for proliferation and differentiation. The second way of apoptosis regulation starts from proliferation stage and performs for 1/2/1 alternations under diminished content of neutrophils and a little increase under next maturation. It leads to resistant depressed apoptosis levels that, at maximal points, are 3-8 times lower than those under alternation 2/1/2. Resistant apoptosis blocking is observed in the Ph+ cells with prolong proliferation or maturation stages, when blasts and myelosytes are accumulated under enhanced bcr/abl and bcl2 > box gene expression and remain under next maturation. Stable apoptosis blocking is accompanied by increasing amounts of blasts and myelocytes and enhancing bcr/abl and bcl 2 > bax expression. This is observed under CML progression. Ph+ cells cultivation may be useful for more distinct diagnostics of CML phases of individual CML patients and optimization of the treatment.

The genes p53, mdm2, p21, c-myc, bcr/abl, bcr, bcl2, bax, and gapdh participate in the regulation of cell proliferation and differentiation, apoptosis and cell distribution for the cell cycle ex vivo in the Ph +cells of chronic myeloid leukemia containing the Ph chromosome and bcr/abl oncogene. Expression of these genes correlates with regulation of cell proliferation and differentiation by alternating proliferation and maturation stages for three main Ph +cell types that occur under chronic myeloid leukemia. The p53, p21, mdm2, and gapdh genes overexpress in active proliferating myeloid cells in the cell cycle S+G2/M phases and when the phases are coincident with the proliferation stage. Expression of these genes decreases to a considerable level under alternation of the Ph +cell proliferation and maturation stages and whenever the expression is greatly diminished under significant neutrophil accumulation and especially under repeated alternation of the stages. In the course of neutrophil maturation, gene expression levels decrease in the range of gapdh actin c-myc, bcr/abl,p21 p53 bcl2 bax. The expression levels of these genes in neutrophils are lower than those in myelocytes and lower by an order of magnitude than that in the cells with a prolonged proliferation stage. The Bcr/abl expression gene under prolonged maturation and neutrophil accumulation is inhibited; however it is enhanced by 2-3 times for the proliferation stage with myelocyte accumulation. Minimal bcr/abl expression is observed under overexpression of p53, mdm2, p21, c-myc, as well as under cell maximum at the S and G2/M phases. Bcr/abl overexpression is observed under low expression of the p53, p21, mdm2 genes. In the Ph + cells with a high P/D efficiency index (5-20), overexpression of the genes in the range of bcr gapdhbcr/abl, as well as a decreased expression of the p53, bcl2, mdm2, p21 gapdh genes is observed for Ph + cells from the CML blast crisis and CML acceleration phase. Low control of cell proliferation and cell cycle by gene-regulators presumably promotes bcr/abl overexpression and activates the production of bcr/abl + cells. Apoptosis in the Ph + cells is induced by expression of the bax bcl2, p53, p21, c-myc and gapdh genes. The blocking of Ph + cell apoptosis, neutrophil accumulation, and decrease in the expression of the p53, mdm2 and p21, c-myc, bcr/abl genes occur at the maturation stage.

Cell regulation of Ph(+)cell proliferation and differentiation has been studied ex vivo in various chronic myeloid leukemia (CML) patients. The regulation is provided by alternation of effective stages of proliferation and maturation with inhibition of Ph(+) cell proliferation by accumulating neutrophils under apoptosis blockage. The alternation of stages consists of switching stage 1 (effective proliferation) to stage 2 (effective maturation) and proceeds according to the 1/2 -1/2/1 or 2/1-2/1/2/1 schemes. The kinetic plots of alternations pass through control points of crossing plots, where the parameters of proliferation and maturation are equal. The indices of P/D efficiency (ratio of proliferation and maturation rates) are 1.06±0.23 and don't depend on time, alternation order, or sources of Ph(+) cells - CML patients. During stages alternation, conversely, the parameters of Ph+ cell proliferation and maturation vary. The proliferation stages are characterized by increased proliferating cells content, a decreased number of neutrophils, and apoptosis induction. At the maturation stages, conversely, apoptosis is inhibited, the number of mature neutrophils increases, while immature Ph(+) cells decrease. High content neutrophils inhibit the proliferation of Ph(+) cells and impair their own maturation by inversion of maturation order, probably through a feedback mechanism. The regulation differences ex vivo reveal three types of Ph(+) cells from various individual CML patients, distinguished by the number and duration of alternating stages of proliferation and maturation. Ph(+) cells types 1 and 2 have one prolonged stage of effective proliferation or effective maturation with efficiency indices P/D(1) = 1-20 or P/D(2) ⇐ 1. At the same time period, the proliferation and differentiation of the Ph(+) cells type 3 proceeds with repeated alternations of stages with P/D(1) = 1-4 or P/D(2) ⇐ 1. Type 1 Ph(+) cells (~20%) were isolated from patients in advanced stages of CML, while Ph(+) cells types 2 and 3 (30 and 50% correspondingly) were isolated from CML chronic phase patients sensitive to chemotherapy.

Aim. To analyse clinical implications of chromosome 8 trisomy in Ph-negative cells of the bone marrow in patients with chronic myeloid leukemia (CML) treated with inhibitors of tyrosinkinases (ITK). Material and methods. A total of 386 patients with CML (chronic phase - 288, acceleration phase - 77) received imatinib (400-800 mg/day). Because of resistance and/or intolerance some patients were switched to ITK II (nilotinib, dasatinib, bozutinib). This study included 8 CML patients (7 in a chronic phase, 1 in acceleration phase) treated with BCR-ABL ITK inhibitors of the first (imatinib) and the second line (ITK-II). The standard cytogenetic examination, on demand - investigation of the interphase nuclei with FISH, in some cases morphological, cytochemical and histological examinations of the bone marrow were made. Results. The existence of a Ph-negative clone with trisomy of chromosome 8 had no negative effect on the course of the disease. The patients showed a stable hematological and cytogenetic response and no need in changing treatment policy. In long-term follow-up Ph-negative clone with trisomy of the chromosome 8 persisted without a clear trend to rise in most patients. Conclusion. Detection of a Ph-negative clone with chromosome 8 trisomy at early stages suggests parallel existence of Ph-positive and Ph-negative clones. None of the patients had myelodisplasia.

В-клеточный хронический лимфолейкоз (В-ХЛЛ) – самый частый вид лейкозов взрослых, остается неизлечимым заболеванием у подавляющего большинства пациентов. Примерно у 1/3 больных оно протекает доброкачественно и редко требует лечения. Продолжительность жизни пациентов этой группы не отличается от общепопуляционной. У 1/3 больных наблюдается агрессивное течение заболевания, причем большинство этих пациентов погибают в течение 5 лет после установления диагноза. У оставшейся части больных заболевание протекает относительно спокойно, однако и их продолжительность жизни не превышает 7–12 лет от момента диагностики.В последние годы мы наблюдаем существенные успехи в терапии В-ХЛЛ, выражающиеся прежде всего в увеличении количества больных, достигших полной ремиссии.

Human myeloid cells with Ph chromosome (Ph+ cells) from chronic myeloid leukemia (CML) in the course of proliferation and differentiation ex vivo are regulated under alternation of cell proliferation and neutrophil maturation stages by consecutive blocking and inducing apoptosis with of neutrophils participation as well bcr/abl, bax and bcl2 genes expression. Apoptosis regulation of three main Ph+ cells types from CML patients depends on alternation sequences of proliferation (1) and maturation (2) cell stages and realized by two ways. The first one is performed by consecutive blocking and inducing apoptosis under 2/1/2 stage alternation. The way is not described early. Neutrophils accumulation correlates with apoprosis blocking. Apoptosis level enhances under neutrophils exhausted. Apoptosis blockage allows cells to proliferate and, thus, to form new portion of neutrophils with consecutive regular their death as well a consequent alternation of apoptosis blocking and inducing. This way regulates proliferation efficiency indexes P/D that reflect Ph+ cells proliferating potential and performs cycle completion for proliferation and differentiation. The second way of apoptosis regulation starts from proliferation stage and performs for 1/2/1 alternations under diminished content of neutrophils and a little increase under next maturation. It leads to resistant depressed apoptosis levels that, at maximal points, are 3-8 times lower than those under alternation 2/1/2. Resistant apoptosis blocking is observed in the Ph+ cells with prolong proliferation or maturation stages, when blasts and myelosytes are accumulated under enhanced bcr/abl and bcl2box gene expression and remain under next maturation. Stable apoptosis blocking is accompanied by increasing amounts of blasts and myelocytes and enhancing bcr/abl and bcl 2bax expression. This is observed under CML progression. Ph+ cells cultivation may be useful for more distinct diagnostics of CML phases of individual CML patients and optimization of the treatment.

To ascertain efficacy of modern treatment methods in idiopathic thrombocytopenic purpura (ITP), to characterize basic indications for administration of these methods; perspective trends in ITP therapy.The article presents 20-year experience in observation and treatment of 1000 ITP patients gained from 1988 by the department of standardization of blood disease treatment methods on the basis of a consultative and outpatient department of Hematological Research Center. The results were assessed by conventional ITP treatment lines.First-line therapy--glucocorticoids--provided remission in 70% patients, long-term (60 months and longer) in 14%, prednisolone resistance was in 19% patients.Intravenous immunoglobulin provided a rapid hemostatic effect (in 1-2 days) in all the patients. A positive response to treatment was seen in 86% patients but it was short-term (for a year and longer in 27%, resistance to the drug--14%). Second line treatment--splenectomy--is the most effective treatment: 80% remission, 32% patients had remission for longer than 60 months. Resistance occurred in 6% patients. Ineffectiveness of treatment in 20% patients stimulated the search for new pathogenetic treatment among synthetic analogues of thrombopoietin. Clinical trials proved high efficacy of two of them--eltrombopag and romiplostim (86-87% response), possible maintenance of remission in long-term interrupted administration of low doses.Modern ITP treatment allows a complete management of hemorrhagic syndrome and deep thrombocytopeny in 80% patients with provision of good quality of life and ability to work. Introduction into clinical practice of thrombopoietin analogues improves treatment results including in 20% patients resistant to all treatments in the absence of marked side effects even in long-term 3-year administration of such medication.

To analyse causes of reactive lymphadenitis regarding histological changes in the lymph node, of diagnostic errors; to determine frequency of establishment of nosological diagnosis.A retrospective analysis of diagnostic examination of 375 patients with reactive lymphadenitis observed from 1992 to 2000 was made.For differential diagnosis 6 groups of reactive lymphadenopathies with different histological picture were identified: follicular hyperplasia, paracortical hyperplasia, granulomatous lymphadenitis, granulomatous pyonecrotic lymphadenitis, sinus histiocytosis as a leading sign and node necrosis as a leading sign. The analysis of making nosological diagnosis for each group and of diagnostic errors was conducted.Characteristics of lymph node changes in reactive lymphadenitis is of great importance. Better cooperation between histologists and clinicians must improve efficacy of reactive lymphadenitis diagnosis.

To evaluate efficacy and tolerance of glivek in chronic myeloid leukemia (CML) in patients who failed interferon-alpha (If-a) preparations.79 patients in a chronic phase of Ph + CML with hematological and cytogenetic resistance or intolerance of If-a. The response to glivec was assessed by achievement of a complete hematological remission and the cytogenetic effect (the degree of reduction of cell clone Ph+ in bone marrow). Tolerance and safety of the drug was studied by monthly standard clinicohematological tests.Not only a hematological remission (92.4%), but also partial (46.8%) or complete (27.8%) elimination of BCR-ABL +/- cells were achieved after 12 months of the treatment. Glivec was well tolerated. Hematological toxicity primarily as neutropenia and thrombocytopenia were observed in 54.4 and 42% patients, respectively. Neutropenia of the third degree which made impossible to continue the treatment was observed in 29.1% patients; throbocytopenia of the third degree was registered in 16.5% patients. Among most frequent non-hematological side effects there were moderate edema, nausea, leg muscle convulsions, weight gain, arthralgias, skin eruption. All the complications were transient, were managed in all cases with only a short-time discontinuation of glivec therapy.High activity of glivec at early stages of CML allows using this drug as a first-line therapy in patients with CML.

Mechanisms of suppressive effect of measles and parotitis virus on lymphocyte proliferation and cytolytic activity of natural killers are studied in vitro. Both viruses exert a weak mitogenic effect on human blood lymphocytes and at the same time suppress their response to phytohemagglutinin stimulation in vitro. Suppressive effect of measles virus is more expressed than of parotitis virus. Moreover, measles virus suppresses lysis of human blood killer target cells. The suppressive effect manifests as early as in 1 h and increases after 3 h of natural killer cell contact with measles virus.

According to the classification of chronic lymphocytic leukemia (CLL) proposed by A. I. Vorob'ev and M. D. Brilliant in 1983, benign CLL is a distinct form of CLL which is characterized by low level of absolute lymphocytosis, absent or mild peripheral lymphadenopathy, slow progression. No specific therapy is needed. The paper presents clinical, morphological and immunological analysis of 34 cases of benign CLL (17 males and 17 females, mean age 58 years). Patients were included in the study if they had lymphocyte count less than 30,000 and no significant growth of lymphoid tissue for at least 3 years. They were followed up from 3 to 24 years (11 years, on the average). The main features of benign CLL are the following: no "B" symptoms, no essential enlargement of the lymphoid organs, a stable low level of absolute lymphocytosis, low prolymphocyte count in the blood smear (0.95% +/- 0.2), nodular or nodular-interstitial proliferation in the bone marrow. We failed to find any cases with paraprotein secretion. There was immunophenotype typical for CLL in 91% of cases (CD19+, CD20+, CD23+, CD5+, EM+, CR1-/CR2+, sIg+(-)). None was positive for CD38 activation marker. One trisomy 12 cases was detected by FISH method. 8 patients died so far, but not because of the tumor progression or transformation, median survival was 22 years.

The paper presents new findings in favor of recognition of splenic lymphocytoma (SLC). This disease was characterized by A. I. Vorob'ev and M. D. Brilliant in 1982 in terms of detailed clinicomorphological features, prognosis and optimal treatment policy. The study included 52 patients (mean age 53 years) of which 36 were females and 16 males. They were followed up for 5.7 years, on the average. SLC manifested clinically by splenomegaly with minimally enlarged lymph nodes, morphologically by nodular lymphocytic proliferates in the spleen, bone marrow and liver, diffuse or diffuse-nodular proliferation in the lymph node. Peripheral blood contained middle-size lymphoid cells with round nuclei. SLC immunophenotype exhibits moderate or marked expression of CD22 and membrane immunoglobulins, the absence of CD5, CD23 and EM receptor, combination of CR1-/ CR2+. Paraprotein secretion was recorded in 49% of cases. There were frequent autoimmune reactions, especially against erythroid cells and platelets (42%). Optimal therapeutic policy is expectation and eventual splenectomy producing a persistent clinical effect in 94% of patients. In progressive disease long-term therapy with cyclophosphamide is recommended. Thus, SLC is a mature-cell lymphatic tumor growing as a rule in the spleen. Its prognosis in valid therapy is favourable.

The examination of peripheral blood lymphocytes from persons exposed to low-dose radiation after the Chernobyl accident demonstrated that the exposure to radiation at the time of the accident and further living in the contaminated territory entail a significant increase in the number of cells with a large thick nucleus and scare cytoplasm. Such morphological picture may be attributed to stable adaptation typical for persistent activation of the general adaptation syndrome and is explained by adaptation-stress relations.

Effects of intravenous and oral hinipral on the uteroplacentofetal blood flow and fetal heart work were studied in tocolytic therapy of preterm labor in 27 pregnant women at terms 25-35 weeks. Intravenous infusions of hinipral resulted in a significant reduction of the systolic-diastolic ratio in the umbilical artery, with the blood flow in the uterine arteries persisting stable, and in a significant reduction of the fetal heart rate and increase of fetal heart rhythm variability, as evidenced by cardiotocogram. In oral tocolysis the reduction of the umbilical artery was related to the pregnancy term; fetal heart rate and heart rhythm variability were within the normal range over the course of the investigation. The results evidence no negative effects of the drug on the uteroplacentofetal blood flow and fetal cardiovascular system both in oral and parenteral forms of tocolysis.

The authors have investigated the possibility of using sera with cold auto-lymphocytotoxic antibodies for the differential diagnosis of chronic lymphocytic leukemia (CLL) and malignant non-Hodgkin's disease (MNHD) at the stage of leukemization. The new rapid immunologic method was approved in the diagnosis of 40 CLL and 52 MNHD patients. The high informative value of this method has permitted the authors to recommend its use as an additional procedure for the differential diagnosis of the above lympho-proliferative diseases.

Comprehensive examination, making use of cardiotocography, dopplerometry, and doppler echocardiography, was carried out to disclose the contribution of fetal hemodynamic disorders to the pathogenesis of antenatal deceleration in 69 pregnant women. Disorders of both intracardiac fetal hemodynamics and of the fetoplacental circulation were revealed in all the cases with deceleration. Therefore, circulatory disorders underlie the pathogenesis of decelerations. The development of decelerations is related to the progressive deterioration of fetoplacental circulation and, specifically, to marked changes in the intracardiac hemodynamics in conditions of augmenting peripheral vascular resistance and hypoxia of the fetus.

Children immunized with live measles vaccine in the foci of measles infection varying in intensity (1-9 cases per focus) have been subjected by two methods: the hemagglutination inhibition (HAI) test and the enzyme immunoassay (EIA). As shown in this study, in most cases (98% of all blood serum samples) the correlation between the results of the HAI test and EIA is not high (r = 0.5), which is linked with the detection of a wider spectrum of antibodies in EIA. The percentage of seronegative children detected by these two methods was practically the same (4.05 and 4.4, respectively). The analysis of the results obtained in this study indicates that EIA is a more informative and sensitive method, which confirms the effectiveness of its use for the determination of the level of collective immunity.

The investigation of adsorption capacity of B- and T-lymphocytes in respect to autolymphocytotoxic antibodies (ALCTA) has shown that these two populations of mature cells have a common antigen against which ALCTA are directed. This antigen is insufficiently expressed of pathologic cells of patients with B-cellular variant of chronic lymphoid leukemia (CLL). B-lymphocytes of patients with malignant non-Hodgkin's lymphoma, hairy-cell leukemia, Waldenström's macroglobulinemia are more mature, as compared to those of CLL patients, and carry surface antigens to which autoantibodies are directed.

The data on the clinical trials of karminomycin, a new antitumor antibiotic are presented. The drug was used in the treatment of 46 adult patients with leukemia. Karminomycin was used in primary inducing therapy and treatment of relapses. The results of the trials showed that karminomycin had a definite therapeutic activity in treatment of acute myeloblast leukemia at various stages of the process. A rapid effect of the antibiotic provided its use in emergency cases with rapidly progressing variants of the disease.