[Endocrine disorders after combined chemoradiotherapy in Hodgkin Lymphoma survivors].

BACKGROUND: Hodgkin's lymphoma (HL) is one of the most common malignant lymphoproliferative diseases. Chemotherapy and radiotherapy used in the treatment of LH induce a number of toxic effects leading to dysfunction of endocrine system. Hormonal disorders in HL and their relationships with the therapy used remain to be clarified. AIM: To assess disorders of the endocrine function of thyroid, parathyroid glands and gonads in HL survivors. MATERIALS AND METHODS: Screening of endocrine dysfunction of the thyroid, parathyroid glands and gonads was performed in 160 adult patients with HL, 55 men and 105 women, at remission stage induced by chemotherapy or chemoradiotherapy. Forty healthy subjects, matched by age, were acted as control. The levels of TSH, T3, free T4, PTH, FSH, LH, free testosterone, dehydroepiandrosterone sulfate (DHEA-S), and sex-hormone binding globulin (SHBG) were measured in blood serum by ELISA. Bone mineral density (BMD) was assessed by DEXA. RESULTS: Hypothyroidism (25%), hyperparathyroidism (15.6%) and hypogonadism (29% of men and 25.3% of women) were the most prevalent endocrine disorders in LH survivors. Hypothyroidism was significantly more common in patients after chemoradiotherapy than in those who received only chemotherapy (χ2=9.4, р=0.002). In patients with hyperparathyroidism, there were negative correlations between PTH levels and BMD in the lumbar spine (r=-0.74, p=0.00002) and in the femoral neck (r=-0.66, p=0.0003). Men with HL demonstrated lower free testosterone concentrations when compared to control (p=0.04); LH and FSH levels were elevated (p=0.0004 and p=0.04, respectively). In men with HL the levels of DHEA-S were reduced (p=0.0009). The increased SHBG concentrations were revealed in 13 (23.6%) men. Women of reproductive age with HL had higher levels of LH in the luteal phase (p=0.05) and FSH in the follicular phase (p=0.02) than controls. CONCLUSION: The data indicate a high prevalence of the dysfunctions of thyroid, parathyroid glands, and gonads in HL survivors. Screening for endocrine disorders in these patients is highly recommended.

[MicroRNA Expression Profiling of Diffuse Large B-Cell Lymphoma].

Non-Hodgkin lymphoma (NHL) is a heterogeneous group of cancers that differ in pathogenesis and prognosis. The main methods of treating NHL include chemotherapy, immunochemotherapy, and radiation therapy. However, a significant proportion of these tumors are chemoresistant or rapidly recur after a short chemotherapy-induced remission. In this regard, the search for alternative cytoreductive therapeutic methods is relevant. Aberrant expression of microRNA (miRNA) is one of the mechanisms responsible for the emergence and progression of malignant lymphoid neoplasms. We analyzed the profile of miRNA expression in the biopsy material from lymph nodes affected by diffuse large B-cell lymphoma (DLBCL). The key material of the study was histological preparations of lymph nodes obtained by excisional diagnostic biopsy and treated using conventional histomorphological formalin fixation methods. The study group consisted of patients with DLBCL (n = 52); the control group consisted of patients with reactive lymphadenopathy (RL) (n = 40). It was shown that the miR-150 expression level in DLBCL was reduced by more than 12 times (p = 3.6 x 10^(-15)) compared with RL. Bioinformatics analysis revealed the involvement of miR-150 in the regulation of hematopoiesis and lymphopoiesis. The data we obtained allow us to consider miR-150 as a promising therapeutic target with great potential in clinical practice.

Study of mRNA of WT1, BAALC, EVI1, PRAME and HMGA2 genes in whole blood samples.

Simultaneous quantitative measurement of mRNA of the WT1, BAALC, EVI1, PRAME and HMGA2 genes in whole blood samples reflects the specific pathological proliferative activity in acute leukemia and their ratio is promising as a diagnostic marker. The transcriptome profile of acute leukemia cells is usually assessed using NGS or microarray techniques after a preliminary procedure for isolation of mononuclear cells. However, the results of using the multiplex PCR reaction for the simultaneous determination of all above mRNAs in whole blood samples have not been published so far. Determination of mRNA of WT1, BAALC, EVI1, PRAME and HMGA2 genes in venous blood level samples by multiplex RT-PCR. The study included 127 blood samples from patients who diagnosis of acute leukemia was subsequently confirmed. In the comparison group, 87 samples of patients without oncohematological diagnosis were selected, including 31 samples (K1) with a normal blood formula and 56 samples (K2) with a violation of the cellular composition - anemia, leukocytosis and thrombocytopenia. RNA isolation and reverse transcription were performed using the Ribozol-D and Reverta-L kits (TsNIIE, Russia). Determination of the mRNA expression level of the WT1, BAALC, EVI1, PRAME and HMGA2 genes by multiplex real-time PCR using a homemade multiplex PCR kit. The mRNA level was characterized by high interindividual variation and did not correlate with the rate of circulating leukocytes or blood blasts. Expression of WT1 mRNA was observed in whole blood only in one patient from the control group and in 112 (88%) patients with leukemia and was combined with a decrease in the level of HMGA2 mRNA expression and BAALC mRNA values. In contrast to the control groups, patients with leukemia had higher levels of BAALC mRNA in AML and ALL, increased PRAME mRNA in AML and APL, but lower levels of HMGA2 in APL.

MicroRNAs in the Myelodysplastic Syndrome.

The myelodysplastic syndrome (MDS) holds a special place among blood cancers, as it represents a whole spectrum of hematological disorders with impaired differentiation of hematopoietic precursors, bone marrow dysplasia, genetic instability and is noted for an increased risk of acute myeloid leukemia. Both genetic and epigenetic factors, including microRNAs (miRNAs), are involved in MDS development. MicroRNAs are short non-coding RNAs that are important regulators of normal hematopoiesis, and abnormal changes in their expression levels can contribute to hematological tumor development. To assess the prognosis of the disease, an international assessment system taking into account a karyotype, the number of blast cells, and the degree of deficiency of different blood cell types is used. However, the overall survival and effectiveness of the therapy offered are not always consistent with predictions. The search for new biomarkers, followed by their integration into the existing prognostic system, will allow for personalized treatment to be performed with more precision. Additionally, this paper explains how miRNA expression levels correlate with the prognosis of overall survival and response to the therapy offered.

[Frequency, spectrum, and functional significance of TP53 mutations in patients with diffuse large B-cell lymphoma].

A comparative analysis of oncogene mutations shows that variations in their frequency, spectrum, and hot-spot locations depends on the type of tumor and the ethnic origin of the population studied. The current version of the IARC TP53 Mutation Database lacks information about the frequency and spectrum of TP53 mutations in patients with DLBCL in Russia. The aim of this study was to assess the frequency and functional significance of TP53 mutations in patients with DLBCL in Novosibirsk. The TP53 coding sequence and the adjacent intron regions were analyzed by direct sequencing in the tumor material from 74 patients with DLBCL. Mutations of the TP53 coding sequence were found in 18 (24.3%) patients. These data are consistent with the frequency of TP53  mutations observed in other studies. The spectrum of nucleotide substitutions found in DLBCL specimens corresponded to that described in the IARC TP53 Mutation Database. According to bioinformatic data and to reported experiments in vitro, most of the mutations detected result in the production of functionally inactive p53. Our results show that DLBCL progression is accompanied by the functional selection for mutations in TP53 exons 5-8.

Role of polymorphic loci in HLA-region rs2647012 and rs805288 in the development of non-Hodgkin's malignant lymphomas in Western Siberia.

The data of genome-wide association analysis suggest that human 6p21.3 chromosomal region (localization of HLA genes) contains polymorphic loci influencing the risk of developing non-Hodgkin's lymphomas. We analyzed association of rs2647012 and rs805288 loci with the risk for non-Hodgkin's malignant lymphomas in the population of Western Siberia. Allele and genotype frequencies were determined in the group of 298 patients and in the control group including 551 individuals. Subgroups of diffuse large B-cell lymphoma (86 patients) and follicular lymphoma (25 patients) were analyzed separately. An association of rs2647012 А/А genotype with increased risk of the disease (OR = 2.78, p = 0.002) was detected in the subgroup of diffuse large B-cell lymphoma.

Analysis of the association of polymorphic loci rs917997 in IL18RAP gene and rs187238 in IL18 gene with the risk for non-Hodgkin's malignant lymphomas in Novosibirsk population.

We analyzed the association of polymorphic variants of rs917997 (G/A) locus in IL18RAP gene and rs187238 (G/C) locus in IL18 gene with the risk of malignant non-Hodgkin's lymphomas in Novosibirsk population. Allele and genotype frequencies of the above loci were determined in patients (243 persons) and control group (371 persons) and compared using χ(2) test. None of the analyzed loci showed statistically significant association with the risk of malignant non-Hodgkin's lymphomas.

[Linkage disequilibrium and haplotypes of rs1042522, rs1625895 and rs17878362 gene TP53 markers in patients with diffuse large B-cell lymphoma].

Association of rs1042522, rs17878362 and rs1625895 markers with non-Hodgkin's lymphoma risk is not well studied. Large number of non-Hodgkin's lymphoma entities, as well as a small effect of each of the polymorphisms on the lymphomas risk, leads to the analyses of these markers in each of histological subtypes of lymphoma and to study the polymorphisms in the haplotype groups. The goal of this work was to analyze the frequency, haplotypes and linkage disequilibrium of rs1042522, rs1625895 and rs17878362 in the patients with diffuse large B-cell lymphoma and in control group. The differences in the structure of LD between rs17878362, rs1042522 and rs1625895 TP53 gene in the population of the Siberian region were shown. Haplotype approach wasmore informative in the analyses of association of the gene TP53 polymorphisms and the diffuse large B-cell lymphomas risk in case-control study than the study of each polymorphism. The association of haplotype wArgG with diffuse large B-cell lymphoma risk, and the protective effect of haplotypes wProG or dupProG were identified. The difference in the effects of the haplotype TP53 was noted depending on the homozygous or heterozygous diplotype.

[Intronic polymorphisms of аntionkogene TP53 in patients with indolent variants of Non-Hodgkin's lymphomas].

The frequency of distribution of alleles and genotypes of single nucleotide substitution G13494A in intron 6 and duplications dup16bp in intron 3 of the gene TP53 in 56 patients with indolent non-Hodgkin's lymphoma and variants in the controls was studied. The increase of the frequency of G-allele and G/G genotype of intron 6 of the gene TP53 in lymphoma patients compared with controls (91 and 84 % vs. 79 (p<0,01) and 63 % (p<0,01), respectively) was determined. It was found that individuals carrying the rare A-allele (i.e., having A/G or A/A genotype), had the risk of disease 3,23 times (OR=3,23; [95 % CI 1,50; 6,92], p<0,05) higher than the general population. No significant differences in the distribution of alleles and genotypes of dup16bp intron 3 of the TP53 gene between cases and controls were identified. The results indicate that the oligonucleotide G13494A substitution in intron 6 proapoptotic gene TP53 in older individuals may have a modulating effect on the risk of indolent lymphoma.

[The stage of portal blood flow in patients with hemoblastosis combined with chronic hepatitis].

109 patients with hemoblastosis (HB) were examined. Average age was 31.8 +/- 1.4 years in group of lymphogranulomatosis patients, and 44 +/- 5.3 years in group of acute leukemia patients. Group of 46 patients with clinical laboratory signs of hepatic lesion was marked out. Since features of portal blood flow (PBF) revealed at dopplerography did not depend on hepatitis aetiology, all patients with chronic hepatitis were analyzed in one group. These patients were examined clinicobiochemically: functional hepatic tests, markers of viral hepatitis B and C, ultrasonic abdominal scanning, PBF dopplerography, puncture biopsy of hepar. All patients had PBF disturbances such as decrease of portal vein blood flow, increase of venous drainage of hepar and spleen. The degree of disturbances intensity increased depending on degree of chronic hepatitis activity (the index of histological activity) and hepatic fibrosis stage. Thus, presence of chronic hepatitis in HB patients regardless of aetiology, is the factor aggravating patients' stage and corrupting PBF. PBF disturbances (decrease of arterial and venous inflow and increase of venous outflow) and development of fibrous changes apparently related with compression of vessels by connective tissue around central veins, inside of lobules and among of hepatic cells. Consequence of it is including anastomosises in blood flow which leads to redistribution of PBF and portal hypertension forming.

[Prognosis factors in imatinib mesilate therapy in patients with a chronic phase of Ph-positive chronic myeloid leukemia: data from a multicenter non-randomized trial in Russia].

AIM: To reveal prognostically significant factors affecting efficacy of glivek therapy in untreated (duration of the disease < or = 6 months) and pretreated (duration of the disease > 6 months) patients with chronic myeloid leukemia (CML) in a chronic phase. MATERIAL AND METHODS: A total of 338 patients (64 untreated and 274 pretreated) with a chronic-phase CML on glivek therapy entered the trial. RESULTS: Five-year survival on glivek was high (89, 98 and 88% in untreated and pretreated patients, respectively). Incidence of transformation in the acceleration phase and blast crisis was low both in untreated and pretreated patients (1.6 and 11%, respectively) and correlated with the rate of a complete cytogenetic response (CCR). Untreated patients had no factors affecting treatment efficacy negatively, CCR probability was 96%. Blastemia, thrombocytosis and splenomegaly reduced CCR probability significantly in pretreated patients. Slow reduction of the tumor mass, late achievement of a complete hematological response and a cytogenetic response decreased probability of CCR. CONCLUSION: Glivek is a drug of choice for patients with chronic-phase CML. High probability of CCR both in untreated and pretreated patients lowers the risk of the disease transformation into the phase of acceleration/blast crisis and raises overall survival in both groups.

Expression of the MDR1 and MRP genes in patients with lymphoma with primary bone marrow involvement.

Expression of MDR1 and MRP genes in patients with low-grade and high-grade non-Hodgkin's lymphomas with primary bone marrow involvement before and after chemotherapy was investigated. The data demonstrate that overexpression of MDR1 and MRP genes in hematological malignancies elevates in patients after chemotherapy and correlates with poor clinic prognosis and more frequent recurrences of the malignancies.

[Morpho-functional condition of heart in patients with lymphogranulomatosis late after chemoradiotherapy]. / Morfofunktsional'noe sostoianie serdtsa pri limfogranulematoze v otdalennye sroki posle khimioluchevoi terapii.

AIM: The study of clinical and echocardiographic status of the heart in patients with lymphogranulomatosis (LGM) late after chemoradiotherapy. MATERIAL AND METHODS: 44 patients with IIA-IV stage of LGM exposed to irradiation of lymph nodes and polychemotherapy according to the schemes ACOP, ABVD, COPP, CHOP, CVPP were examined. Echocardiography was carried out on the unit Sigma-44 and Toshiba. RESULTS: Some changes in the heart by type of myocardiodystrophy or endomyocardial fibrosis were found. The latter are characterized by diminution of the left and (or) right ventricles due to apex obliteration, hardening and thickening of the endocardium and subvalvular structures, diastolic dysfunction and pulmonary hypertension. The main and additional signs are distinguished. The reasons of endomyocardial fibrosis are discussed: severity of the disease, frequent exacerbations and, consequently, higher doses of specific chemoradiotherapy. CONCLUSION: It is necessary to employ sparing policy in planning radiotherapy after high loading antracycline antibiotics.

[Ultrastructural changes in the liver cells in severe iron-deficiency anemia]. / Ul'trastrukturnye izmeneniia v kletkakh pecheni pri zhelezodefitsitnoi anemii tiazheloi stepeni.

Ultrastructural changes in hepatocytes, sinusoidal endothelial and Kupffer cells and their phagocytic activities were studied in rabbits with grave posthemorrhagic iron deficiency anemia. Morphologic characteristics of depressed mitochondrial function and cellular plastic processes were observed in all liver cells. Necrobiotic changes in endothelial and Kupffer cells were discovered. Kupffer cells phagocytic capacity was decreased, this being shown by latex particles phagocytosis. The share of secondary lysosomes with more labile membranes increased 2-3-fold, this being a risk factor in the treatment of iron deficiency anemia with lysosomotropic agents, including iron-containing ones.