[Clinical Significance of FGFR1 Gene Abnormalities in Blood Tumors].

OBJECTIVE: To study the potential significance and clinical application of FGFR1 gene abnormality in the diagnosis, clinical features, pathological mechanism and treatment in hematological tumors. METHODS: Clinical data of total of 29 patient with chromosome of 8 short arm (8P) abnormality who had more comprehensive medical history from 2013 to 2018 were collected. The karyotype analysis of bone marrow chromosomes in patients was carried out by using chromosome R band banding technique. FGFR1 gene was detected by using fluorescence in situ hybridization (FISH). RESULTS: Seven cases of FGFR1 gene abnormalities were decteted, including 3 cases of FGFR1 gene amplification, 2 cases of translocation, and 2 cases of deletion. Five patients with FGFR1 gene amplification or deletion not accompaned with eosinophilia, moreover the chromosome was a complex karyotype with poor prognosis; Two cases of FGFR1 gene translocation were non-complex chromosomal translocation and one of which survived for 6 years after bone marrow transplantation, the other chromosome karyotype showed no rearrangement of 8 short arm. However, FGFR1 gene rearrangement was confirmed by FISH analysis, which was a rare insertional translocation. CONCLUSION: FGFR1 gene amplification or deletion often occur in cases with complex karyotype, which not accompany eosinophilia, moreover have poor prognosis. The patients with FGFR1 gene translocation accompany eosinophilia which is consistent with the clinical characteristics of myeloid / lymphoid neoplasms with FGFR1 abnormality. Karyotype analysis combined with FISH method can improve the detection of abnormal clones.

Fusion of platelet-derived growth factor receptor ß to CEV14 gene in chronic myelomonocytic leukemia: A case report and review of the literature.

Myeloid tumor possessing platelet-derived growth factor receptor ß (PDGFRß) gene rearrangement is a rare hematological malignancy, which presents with typical characteristics of myeloid proliferation disorders and eosinophilia. In the present study, an elderly chronic myelomonocytic leukemia patient was diagnosed with chromosome rearrangement. Fluorescence in situ hybridization (FISH) was conducted with a PDGFRß isolate probe, and gene translocation between PDGFRß on chromosome 5 and genes on the chromosomes of group D (13-15) was detected. Karyotype analysis revealed a chromosome 5 break, and PDGFRß-thyroid hormone receptor interactor 11 (CEV14) gene fusion was confirmed via reverse transcription-polymerase chain reaction (RT-PCR), which additionally revealed the chromosome rearrangement t(5;14)(q33;q32). Due to the correlation between PDGFRß-CEV14 expression and effectiveness of treatment with tyrosine kinase inhibitors, this fusion gene is considered to be an oncogene. In the present study, an elderly patient was diagnosed with a myeloid tumor associated with the fusion gene PDGFRß-CEV14, using the methods of FISH and RT-PCR. These methods were confirmed to be of significant value in improving diagnosis, guiding treatment and increasing the cure rate of patients, due to their ability to detect multiple rearrangement genes associated with PDGFRß in myelodysplastic and myeloproliferative neoplasms.

[Clinical and laboratory characteristics of hematological diseases associated with eosinophilia].

OBJECTIVE: To analyze the clinical and laboratory characteristics of hematological diseases associated with eosinophilia. METHODS: Karyotype analysis was performed by direct method and/or short-time culture of bone marrow cells for R-banding. Fluorescence in situ hybridization (FISH) was performed using PDGFRα, PDGFRß and FGFR1 break-apart probes. RESULTS: The clinical and hematological findings of 44 patients were diagnosed as hematological diseases associated with eosinophilia. Abnormal karyotypes were detected in 6 cases (13.64%) with karyotyping. The efficiency of the detection of abnormal clone was markedly increased to 29.55% (13/44) with FISH techniques, including 7 cases with FIP1L1-PDGFRα (F/P, 15.91%), 3(6.82%) PDGFRα rearrangement, 2 (4.55%) aberrant PDGFRß gene and 1(2.27%) FGFR1 rearrangement. Patients being PDGFRα, PDGFRß or FGFR1 positive (13 cases) or negative (31 cases) showed predominant difference in clinical and laboratory features. The incidence of gut involvement, the absolute count of eosinophils in peripheral blood and the percentage of immature eosinophils in bone marrow were significantly increased in positive patients (P < 0.05). CONCLUSIONS: The hematological diseases associated with eosinophilia are characterized by unique clinical and laboratory features. Karyotyping should be a routine approach to detect the abnormal clone in these diseases. Screening for PDGFRα, PDGFRß and FGFR1 gene with FISH can provide more genetic information.

[Clinical and laboratory study of myleodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) with PDGFRß abnormalities].

OBJECTIVE: To explore the clinical and laboratory characteristics of myleodysplastic syndrome (MDS)/myeloproliferative neoplasm (MPN) with PDGFRß abnormalities. METHODS: Chromosome specimens were prepared directly and/or short-time culture of bone marrow cells. Karyotyping was performed with R-binding technique. Fluorescence in situ hybridization (FISH) was performed using PDGFRß, PDGFRα, FGFR1 break-apart probes and whole chromosome 5 and 12 painting probes, respectively. The expression of JAK2 V617F was measured with quantitative PCR. RESULTS: The clinical and hematological findings of 27 patients were compatible with diagnosis of MDS/MPN. PDGFRß rearrangement was detected in 4 patients with D-FISH, and 2 of which were confirmed as t(5;12) by chromosome painting. PDGFRα, FGFR1 and JAK2 V617F mutation were not detected in these 4 PDGFRß positive MDS/MPN patients with. CONCLUSIONS: PDGFRß gene rearrangement may be detected in some MDS/MPN patients. FISH is a convenient and reliable approach to detect PDGFRß gene.

[A novel single nucleotide polymorphism-based method for quantitative assessment of chimerism after allogeneic stem cell transplantation.].

OBJECTIVE: To develop a novel single nucleotide polymorphism (SNP)-PCR based method for quantitative detection of chimerism after allogeneic haemopoietic stem cell transplantation (allo-HSCT), and to explore its feasibility, accuracy and superiority. METHODS: 18 SNP loci were sereened to identify informative markers for detecting chimerism in each donor/recipient pair before transplantation. Then the chimerism rate of each informative marker was analyzed by real-time quantitative PCR (RQ-PCR). The accuracy and sensitivity were verified by multiple proportion dilution and analogy chimerism compared with quantitative detection of short tandem repeat (STR)-PCR, fluorescence in situ hybridization (FISH) and fusion gene. RESULTS: (1) The average slope of the 17 time amplications of the internal control plasmid was -3.39, the average intercept was 39.97, correlation coefficients were more than 0.995, which was close to the theoretical level. The intra- and interassay variability was 0.50% and 1.1%, respectively, which were both in the allowed ranges. A linear correlation with artificial mixed chimerism is above 0.99 and a sensitivity of 0.01% proved reproducible. (2) At least one informative marker could be found in over 95% of 40 donor/recipient pairs. The results of the chimerisms derived from SNP-PCR were consistent with that from STR-PCR (96.7%), FISH and fusion gene analasis (P > 0.05); the quantitative results of special fusion gene transcripts were negtive in complete chimerism samples, and positive in mixed chimerism samples. CONCLUSIONS: This new assay which overcome the PCR competition and plateau biases of STR-PCR provides an accurate, reliable and rapid quantitative assessment of mixed chimerism after allo-transplantation. It is highly promising for of clinical application and may take the place of STR-PCR in the conventional chimerisim assessment.

[An analysis of prognostic factors in 80 myelodysplastic syndrome].

OBJECTIVE: To investigate the WHO classification, clinical and hematological features and risk group of International Prognostic Scoring System (IPSS) in patients with myelodysplastic syndrome (MDS). METHODS: The diagnosis and classification of MDS patients were defined according to the WHO classification. The clinical manifestations, hemogram, bone marrow biopsy and prognosis were retrospectively analyzed. RESULTS: The median age at diagnosis of MDS was 47 yrs being younger than that in some foreign reports. The frequency of abnormal karyotype was 35.14% and +8 was the most frequent abnormal karyotype in our study. Eleven of 74 patients transformed into leukemia. Univariate analysis showed that age, chromosome abnormality, percentage of bone marrow blast cells and number of cytopenias were significantly related to prognosis. There was a statistical difference in cum survival rate between IPSS subcategories (P < 0.05) except that between low- and intermediate I-risk subcategory (P > 0.05). There were statistical differences for refractory anemia (RA) vs RA with excess blast (RAEB), refractory cytopenias with multilineage dysplasia (RCMD) vs RAEB and RAEB-I vs RAEB-II (P < 0.05). CONCLUSIONS: There were differences in age of disease onset, distribution of WHO, sub-classification and abnormal karyotype in this cohort of MDS patients as compared with those in Europe and Japan. It is helpful in diagnosis, treatment and prognosis to divide RAEB into RAEB-I and RAEB-II. IPSS was well applicable in Chinese MDS patients.

[Enrichment and biological characteristics of murine mesenchymal stem cells].

The study was aimed to isolate and establish mesenchymal stem cell line from adult murine bone marrow as well as to identify its biological characteristics and differentiation potential. Bone marrow cells (BMCs) were collected by flushing the femurs and tibias of 4 - 5-week-old male C57BL/6 mice, and were inoculated at a concentration of 1 x 10(6)/cm(2). mMSCs were isolated, enriched and expanded by using bone marrow adherant culture and monoclonal culture. The characteristics of the cells, such as morphology, growth pattern, cell cycle, phenotype, karyotype and multipotent differentiation potential, cytogenetic stability and tumorigenesis were determined. The results indicated that the cell population consisted of spindle- and star-shaped cells, they were highly positive for CD29, CD44, Sca-1, MHC-I, moderate positive for CD13, CD90.2 and negative for CD117, CD45, Flk-1 and MHC-II. mMSCs could be induced to differentiate into adipocytes, osteoblast cells and chondrocytes. It is concluded that mMSC can be isolted, expanded and enriched by using bone marrow adhcrent culture and monoclonal culture. No tumor formations are observed for 3 months in nude mice after subcutaneous injection. mMSCs retain their properties after at least 30 passages in culture as well as from frozen stocks.

[Clinical and laboratory characteristics of two acute lymphoblastic leukemia patients with dicentric (9; 20) (p11 - 13; q11)].

OBJECTIVE: To explore the morphologic, immunophenotypic, cytogenetic and clinical features of acute lymphoblastic leukemia (ALL) patients with dicentric (9; 20) (p11 - 13; q11). METHODS: Chromosome specimens of bone marrow cells were prepared by direct method and/or short-time culture. Karyo-typing was performed by R-banding technique. Dual-color fluorescence in situ hybridization (FISH) was performed using both chromosome 9 classical satellite probe and chromosome 20 alpha-satellite probe in one patient. RESULTS: The two ALL patients were positive for CD10 and HLA-DR, showing of B cell origin. Both patients had dicentric (9; 20): case 1 was 45, XY, der (9) t (9; 20) (p11; q11), -20[20]; case 2 was 45, XX, der (9) t (9; 20) (p13; q11), t (9; 22) (q34; q11), -20[10]/46, idem, +8[16]/47, idem, +8, +21[14]. Mutual translocation between chromosomes 9 and 20 of the dicentric chromosome was confirmed by FISH in one patient. CONCLUSIONS: Dicentric (9; 20) (p11 - 13; q11) is a rare recurring chromosome abnormality associated with ALL. Because of the subtle nature of the translocation, FISH is essential for the detection of this abnormality.

[Report of a case of hybrid acute leukemia with t (12; 22) and literature review].

OBJECTIVE: To report a hybrid acute leukemia (HAL) patient with t (12; 22) (p13; q12). METHODS: Chromosome specimens were prepared by direct method and/or short-time culture of bone marrow cells. Karyotyping was performed by R-banding technique. Leukemia surface markers were detected by anti-biotin-biotin complex and monoclonal antibodies. Chromosome painting (fluorescence in situ hybridization, FISH) was performed by using whole chromosome 12 and 22 probes labeled with green and red fluorescence, respectively. RESULTS: The clinical and hematological findings were compatible with the diagnosis of HAL. Lymphoid and myeloid markers were positive on the leukemia cells. Karyotype analysis showed that the patient had t (12; 22) (p13; q12) translocation. A reciprocal translocation between chromosomes 12p and 22q was proved by FISH. CONCLUSIONS: t (12; 22) translocation is a rare chromosome abnormality in leukemia. Patients with t (12; 22) had unique clinical, cytogenetic features. This translocation as a cytogenetic marker for poor-prognosis in leukemia needs to be further studied.

[Clinical and experimental study of two cases of myelodysplastic syndromes with double isochromosome 20q- anomaly].

OBJECTIVE: To explore the clinical and laboratory characteristics of two myelodysplastic syndromes (MDS) patients with double isochromosome 20q- anomaly. METHODS: Bone marrow cell chromosome preparations were made with both direct method and short-term culture. Karyotype analysis was performed by R-banding technique, and dual-color FISH (fluorescence in situ hybridization) by using a 20q telomeric probe and a sequence-specific probe for 20q12. RESULTS: The clinical and hematological findings were comparable with diagnosis of MDS. Karyotype analysis showed that both patients had double isochromosome 20q- anomaly: case 1 is 46, XX, der(20)? i(20q-) [6]/46, idem, der (6) i (6p) [1]/47, idem, +der (20)? i (20q-) [3]/47, idem, der(6)i (6p), +der(20)? i (20q-) [20]; case 2 is 45, XY, -7, der (20)? i (20q-) [17]/46, idem, +der(20) ? i(20q-) [3]. Two derivative chromosomes 20 were proved 20q isochromosomes with interstitial deletions by dual-color FISH in one patient. CONCLUSIONS: Double isochromosome 20q- anomaly is a rare recurrent karyotype abnormality in MDS, and signals a poor prognosis.

[Clinical and molecular cytogenetic features of myeloid diseases characterized by i(20q-): a study of seven cases].

OBJECTIVE: To investigate the clinical and molecular cytogenetic features of myeloid diseases characterized by i(20q-). METHODS: The clinical data of 7 patients with myeloid diseases, 6 with myelodysplastic syndrome (MDS) and one with acute myelocytic leukemia (AML), 4 males and 3 females, aged 51 - 74, were analyzed. Chromosome specimens were prepared by direct method and/or short-time culture of bone marrow cells. Karyotyping was performed by R banding technique. Two kinds of probes (20q subtelomere probe and 20q12 unique sequence probe) were used in dual-color fluorescence in situ hybridization (D-FISH) assay. RESULTS: Of the seven patients, 4 died and 3 survived by the end of this study. The patients survived for 6 months (case 1), 7 months (case 2), 17 days (case 4), and 28 days (case 5) respectively. Karyotype analysis showed that one of the normal chromosomes 20 was lost and substituted by one or two small metacentric isochromosomes smaller than the normal chromosome 20 in all these seven cases. It was proved to be ider(20)(q10) del(20)(q11q13), i(20q-) in six cases by D-FISH assay. CONCLUSION: i(20q-) is a novel and rare recurrent chromosome abnormality which may be specifically associated with myeloid diseases and poor prognosis.