Adolescent ; Adult ; Aged ; Female ; Humans ; Janus Kinase 2 ; genetics ; Karyotyping ; Male ; Middle Aged ; Mutation ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; Young Adult ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVETo explore the prevalence of IDH gene (IDH1 and IDH2) mutations, types of mutations in patients with acute myeloid leukemia (AML), correlation with the internal tandem duplication(ITD) mutation of FLT3 gene, NPM1 gene mutation and some clinical characteristics.

METHODSThe mutations of IDH1 and IDH2 gene at exon 4, NPM1 gene at exon 12 and FLT3-ITD at exon 14 and 15 in 163 newly diagnosed AML patients were detected by PCR amplification followed by direct sequencing of genomic DNA.

RESULTS(1) IDH mutations were found in 25 patients (25/163), and all were heterozygous, of which IDH1 in 7 patients (4.29%) and IDH2 in 18 (11.04%). A total of 4 types of IDH1 mutations were identified (c.395G→A, p.R132H, n = 4; c.394C→A, p.R132S, n = 1; c.394C→G, p.R132G, n = 1; c.315C→T, n = 1). The IDH1 mutation caused substitutions of residue R132 except for one (c.315C→T). All IDH2 mutations caused changes of R140 (c.419G→A, p.R140Q, n = 18). The incidence of IDH2 mutation was significantly higher than that of IDH1 mutation (11.0% v 4.3%, P = 0.022). Both IDH1 and IDH2 mutation were detected in one patient, while IDH1 was synonymous substitution (c.315C→T). IDH-mutated cases showed a significantly higher frequency of concurrent FLT3-ITD mutation compared with wildtype cases (34.6% vs 11.9%, P = 0.003), so did IDH mutations concurrent NPM1 mutation vs NPM1 wildtype (28.1% vs 12.7%, P = 0.033), of which the frequency of concurrent NPM1 and FLT-ITD mutations cases with the IDH mutation was significantly higher than that of NPM1 and FLT-ITD negative (45.5% vs 11.7%, P = 0.002). IDH mutation incidence was significantly higher in normal karyotype cases than in abnormal ones (20.5% vs 5.8%, P = 0.020). Patients with IDH mutations were significantly older than wildtype patients(P < 0.001), whereas, there were no statistically significant differences in gender, peripheral blood (PB) count at diagnosis between two groups.

CONCLUSIONSThe incidence of IDH mutation is higher in patients with de novo AMLs, of which IDH2 mutation more frequently, and the patients associated with older age, normal karyotype at diagnosis. IDH mutation has a strong association with NPM1 and FLT3-ITD mutations, suggesting that IDH mutation has synergistic effect with the latter gene on leukemogenesis.

Adolescent ; Adult ; Aged ; DNA Mutational Analysis ; Female ; Genotype ; Humans ; Isocitrate Dehydrogenase ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Young Adult

Objective To analyze the correlation between the expressing level of bradykinin B2receptor and pathological grade of gliomas, and supply experimental basis for clinical application of bradykinin or its analog. Methods The clinicopathologic findings were diagnosed by reviewing all hematoxylin and eosin (H&E) stained tissue sections. To determine the expressing level of bradykinin B2 receptor in glioma, immunohistochemistry and Western blot methods were used.Results Results of H&E staining: In 26 cases of glioma there were 9 cases of grade Ⅰ, 9 cases of grade Ⅱ, 8 cases of grade Ⅲ, and 0 case of grade Ⅳ. Bradykinin B2 receptor localized on tumor cells while the cells of edematous band at the edge of glioma did not express B2 receptor. Results of Western blot: grade Ⅰ and grade Ⅱ (39480.88 ± 5119.96 vs 51354.25 ±6168.77, n = 8, t = 2. 447, P ＜ 0.05); grade Ⅰ and grade Ⅲ (39480.88 ± 5119.96 vs 73032.13 ±8802.71, n = 8, t = 7. 710, P ＜ 0. 001 ); grade Ⅱ and grade Ⅲ (51354.25 ± 6168.77 vs 73032.13 ± 8802.71, n = 8,t = 5. 704, P ＜ 0. 001 ). There was a positive correlation between the expressing level of bradykinin B2 receptor and the the pathological grade of glioma (r =0. 895, P ＜0.001) as grade Ⅰ＜grade Ⅱ＜grade Ⅲ. Results of immunohistochemistry:grade Ⅰ and grade Ⅱ (3.54% 1.51% vs 8.47% ±3.45%, n=8, t=3.698, P＜0.01); grade Ⅰ and grade Ⅲ (3.54%±1.51% vs 15.94% ±1.68%, n=8, t=15.562, P＜0.001); grade Ⅱ and grade Ⅲ (8.47% ±3.45% vs 15.94%± 1.68% , n = 8, t = 5. 505, P ＜ 0. 001 ). There was a positive correlation between the positive position area ratio of bradykinin B2receptor and the pathological grade of glioma (r = 0. 878, P ＜0. 001 ) as grade Ⅰ＜ grade Ⅱ＜ grade Ⅲ. Conclusion There was a positive correlation between the expressing level of bradykinin B2 receptor and the pathological grade of glioma. This may be the reason of the diverse cure effects of the increase of blood-tumor barrier permeability induced by bradykinin and its analog.

OBJECTIVE: To investigate the mutation of RUNX1 gene in patients with myelodysplastic syndrome (MDS) and its correlation with other gene mutations and some clinical parameters. METHODS: The mutations of RUNX1, DNMT3A, TET2, IDH1/2, NPM1, FLT3-ITD and C-KIT in 170 patients with MDS were detected by direct and indirect sequencing of genomic DNA-PCR amplification products. RESULTS: The RUNX1 mutation was found in 23 patients (13.5 %, 23/170). Among the 170 patients, other most frequent mutation was TET2 (11.2%, 19/170), followed by mutations in DNMT3A (9.4%, 16/170), NPM1 (8.2%, 14/170), IDH2 (4.1%, 7/170)、FLT3-ITD (2.9%, 5/170), IDH1 (1.7%, 3/170) and c-KIT (0.58%, 1/170). The most common coexisting mutations were TET2 (5/23). The RUNX1-mutated group showed significantly higher leukocyte levels, higher percentages of blast cells, higher incidences of leukemia transformation and lower platelet counts in comparison with RUNX1 non-mutation group (P＜0.05). whereas there were no statistically significant difference in age, MDS subtype, karyotype and hemoglobin level between 2 groups (P＞0.05). Seventeen patients harboring RUNX1 mutations were followed up and almost 47.05% (8/17) of the patients progressed into acute myeloid leukemia (AML). The rates of transformation into AML in ASXL1-mutation group was significantly higher than that in ASXLL- non-mutation group (47.05% vs 11.7%) (P=0.001). CONCLUSION The incidence of RUNX1 mutation is high in MDS patients. The RUNX1-mutated patients have higher leukocyte level, higher percentages of blast cells, higher incidences of leukemia transformation and lower platelet count.