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.

Cultivated ginseng in the farmland would become the mainly planting mode of Panax ginseng. However,there are relatively few cultivation ginseng varieties for farmland in China. Correlative analysis of qualitity and agronomic traits of P. ginseng cultivation in the farmland could provide a reference for the selection of excellent germplasm and new variety breeding of P. ginseng. In this study,the main index of saponin and agronomic traits of 4-6 years' samples were analyzed by UPLC and measured. The results show that there was significant difference in agronomic indexes of Damaya. The coefficient of variation of the root length( CV = 41. 97%) and fresh weight( CV = 31. 81%) were maximum,and the coefficient of variation of the stems thickness( 16. 72%) and root thickness were minimum. There was a significant correlation between yield and root thickness( P<0. 05). There was significant difference in drug yield of different harvest years( P<0. 05),and the yield of 6-years was 31. 52%-39. 69% higher than 4-years. However,there wasn't significant difference in total ginsenosides between 4 and 6 years old P. ginseng,but there was significant difference in ginseng Rg2,Rc and Rb2( P<0. 05),and the ginsenoside contents of different harvesting years were accorded with the criterion standards of 2015 Chinese Pharmacopoeia. There was no significant correlation between the saponin and the agronomic trait,while there was positive correlation with root thickness( P < 0. 05). Therefore,the stem diameter was positive correlation with yield of P. ginseng. Selection of the stem thickness of seedlings is beneficial to the increase of the yield and breeding of P. ginseng.
China ; Crop Production ; Ginsenosides/analysis* ; Panax/chemistry* ; Plant Breeding ; Plant Roots/growth & development* ; Plant Stems/growth & development*

OBJECTIVETo analyze the CARL gene mutation in the patients with chronic myeloproliferative neoplasm(MPN) and to explore the clinical significance of CALR mutation.

METHODSThe peripheral blood of patients was collected and the genomic DNA was exacted, the 9 exon of CALR gene and the fragment of human thrombopoetic receptor(MPL) gene were amplified by PCR, the mutation of CALR and MPL genes was detected by using the direct sequencing, the JAK2 V617F mutation was detected by using allele spicific PCR.

RESULTSThe CALR mutations were detected in 13 patients out of 55 MPN patients (23.6%). The frequency of CALR mutation was 22.7% (10/44) in 44 essential thrombocythemia(ET) patients. A total of 3 types of CALR mutation were identified (type I c.1092_1143del52bp, n=5; type II c.1154_1155insTTGTC, n=4; type III c.1094_1139del46bp, n=1). CALR mutations occurred at a frequency of 27.2% in primary myelofibrosis (PMF), including type I (n=2) and type II (n=1). The incidence of JAK2 V617F was 58.1%(32/55), that in ET and PMF was 59.1%(26/44) and 54.5% (6/11), respectively. The mutations of MPL W515 were not detectable in all cases, and the simultaneous mutation of CARL and MPL W515 was not detected. The median age of patients with CALR mutation was significantly younger than that of patients with JAK2 mutations (48 vs 64 years of old, P<0.05). The levels of hemoglobin and leukocytes in patients with CARL mutations were significantly lower (P<0.05) but the level of plateletes was higher than that in patients with JAK2 V617F mutations (P<0.05). Deep venous thrombosis occurred in 4 of 35 ET patients with the JAK2 V617F mutation (n=4), but did not occurr in the patients with CALR mutation. Karyotype abnormality was detected in only one case among 48 patients by chromosome karyotype analysis.

CONCLUSIONThe incidence of CALR mutation is high in ET and PMF patients without JAK2 V617F and MPL W515K mutations, which is associated with younger median age, lower leucocyte and hemoglobin levels, higher platelet counts, and rare thrombocytosis, compared with the patients with JAK2 V617F mutation.

OBJECTIVETo explore the coexistence of ASXL1 and CALR gene mutations in patients with essential thrombocytheima (ET) and with primary myelofibrosis(PMF), and to compare the differences of clinical characteristics between ET and PMF patients carrying ASXL1 and CALR mutations, and ET and PMF patients carrying solitary gene mutation, and ET and PMF patients without any mutations.

METHODSThe mutations of ASXL1 gene at exon 12, CALR gene at exon 9 and MPL gene at exon 10 in 263 essential ET patients and 29 PMF patients were detected by PCR amplification followed by direct sequencing of genomic DNA. The JAK2V617F mutations were used by allele specific PCR detection.

RESULTS72.6%(212/292)of patients harbored at least one mutation. The incidences of ASXL1 and CALR mutations were 5.8% and 30.5%, respectively. The frequencies of JAK2V617F and MPL mutations were 39.0% and 2.4%, respectively. 5.1%(15/292) of patients had double mutations, including ASXL1 and CALR(n=11), ASXL1 and JAK2V617F(n=2), MPL and CALR(n=1) and ASXL1 and MPL(n=1). The frequency of concurrent ASXL1 and CALR mutations was found to be high. Significant difference was found on hemoglobin levels and platelet counts between CALR and ASXL1 mutations and single mutation (P<0.05),however, the difference on leukocyte counts and median age was not found. Compared with negative patients, the presence of ASXL1 and CALR mutations was found to be significantly correlative with lower hemoglobin level (P=0.045), lower leukocyte count (P=0.002) and with higher platelet counts(P=0.001), but the difference of median age was not found.

CONCLUSIONThe frequency of concurrent ASXL1 and CALR mutations is higher in ET patients. The coexistence of ASXL1 and CALR gene mutations significantly associated with lower hemoglobin level and higher platelet count.

Objective To investigate the value of chloride clearance test in differential diagnosis of Gitelman syndrome (GS). Methods For patients with hypokalemic metabolic alkalosis and highly suspected GS,clinical data were documented and SLC12A3 gene screening was performed as gold standard to diagnose GS. Hydrochlorothiazide (HCT) test and furosemide (FUR) test were performed according to the standard process. Baseline and maximal increasement of chloride excretion fraction (FECl,the net and relative increase measured as εFECl) were compared between patients and controls to evaluated the reaction to the corresponding diuretics. Receiver operating characteristic (ROC) curve was used to evaluate the sensitivity and specificity of HCT test in GS diagnosis. Results Totally 27 patients and 20 health controls received HCT test. Among those patients,23 were diagnosed with GS genetically. When using the net and relative εFECl to diagnose GS,the areas under the ROC curve were 0.987 (95% CI:0.963～1.000,P<0.001) and 0.984 (95%CI:0.950～1.000,P<0.001),respectively. When a reasonable cutoff value for εFECl was selected,the sensitivity and specificity were both higher than 95%. Eight patients received both HCT test and FUR test. Five of them showed decreased reaction to HCT(net εFECl≤2.86% or relative εFECl≤223%),while normal reaction to FUR.SLC12A3 mutations confirmed their GS. Three patients with blunt reaction to FUR showed normal reaction to HCT,finally they were diagnosed as BS clinically because no SLC12A3 gene mutation was detected. Conclusion Comprehensive application of HCT test and FUR test to evaluate the diuretic reaction can effectively differentiate GS and BS.
Case-Control Studies ; Chlorides ; metabolism ; Diagnosis, Differential ; Gitelman Syndrome ; diagnosis ; Humans ; Hydrochlorothiazide ; Kinetics ; Mutation ; ROC Curve ; Sensitivity and Specificity ; Solute Carrier Family 12, Member 3 ; genetics ; metabolism

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

OBJECTIVETo analyze the correlation between clinical features and cytogenetic finding of 45 adult patients with acute lymphoblastic leukemia (ALL), and to assess the value of chromosomal examination for the diagnosis and prognosis.

METHODSFluorescence in situ hybridization (FISH) was utilized for detecting the BCR/ABL fusion gene and P53 gene. Median survival time (MST) of patients was compared using Log-rank test.

RESULTSRespectively, the MST of patients with white blood cell count (WBC) ≤30 × 10(9)/L, normal karyotype, or without a Philadelphia chromosome were significantly greater than those with WBC > 30 × 10(9)/L, abnormal karyotype or Philadelphia chromosome (P< 0.05).

CONCLUSIONWBC, karyotype abnormalities and presence of Philadelphia chromosome are independent factors for the prognosis of ALL in adult patients.

Abnormal Karyotype ; Adult ; Aged ; Cytogenetic Analysis ; methods ; Female ; Fusion Proteins, bcr-abl ; genetics ; Genes, p53 ; Humans ; Male ; Middle Aged ; Philadelphia Chromosome ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; diagnosis ; genetics

OBJECTIVETo investigate the prognostic value of t(11; 18) (q21; q21) in gastric mucosa-associated lymphoid tissue lymphoma.

METHODSA cohort of thirty-six gastric mucosa-associated lymphoid tissue lymphoma patients who were pathologically identify diagnosis from January 1994 to June 2004 were followed up retrospectively and studied using fluorescence in situ hybridization(FISH) technique to detect t(11; 18) (q21; q21) chromosomal translocation on preservative paraffin specimen.

RESULTSAmong thirty-six patients, fifteen (41.67%) were positive for t (11; 18) (q21; q21). All but one were followed up to March 2010, general median survival time (MST) was 87 months. The MST were 43 and 130 months for t(11; 18) positive and negative patients, respectively. The MST between these two groups was notably different (chi-square=29.57, P< 0.01).

CONCLUSIONt(11; 18) (q21; q21) is important prognostic factor for gastric mucosa-associated lymphoid tissue lymphoma.

Adult ; Aged ; Aged, 80 and over ; Chromosomes, Human, Pair 11 ; Chromosomes, Human, Pair 18 ; Cohort Studies ; Female ; Follow-Up Studies ; Gastric Mucosa ; pathology ; Humans ; Lymphoma, B-Cell, Marginal Zone ; genetics ; pathology ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Translocation, Genetic

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

The aim of this study was to detect the expressions of transforming growth factor (TGFβ(1)), tumor necrosis factor alpha (TNFα) and leukemia inhibitory factor (LIF) in newly diagnosed patients with acute myeloid leukemia (AML) and investigate the association between serum levels of various cytokines and clinical outcomes. The levels of TGFβ1, TNFα and LIF in patient's plasma were detected by enzyme-linked immunosorbent assays (ELISA) and were compared with healthy controls; bone marrow cell morphology, immunology, cytogenetics examinations (MIC) were performed meanwhile. The results showed that levels of TGFβ1, TNFα and LIF were elevated in AML patients as compared with the controls (13.08±9.77 ng/ml, 10.67±15.11 pg/ml, 4.23±4.73 pg/ml vs 8.23±3.12 ng/ml, 5.86±3.05 pg/ml, 2.78±1.22 pg/ml) (p all<0.05). The three cytokines and MIC examination analysis indicated that level of LIF was abnormally elevated in M5 patients (7.14±6.62 pg/ml); TNFα was abnormally elevated in M4 and M3 patients especially M4; TGFβ1 level in M6 and M2 patients was higher than others. TGFβ1 plasma concentration in low-risk group the lowest (10.45±4.73 ng/ml), and that in middle risk group was the highest (16.13±13.76 ng/ml) (p<0.05); the levels of other two kinds of factors in the chromosome karyotype groups showed no significant difference. It is concluded that TGFβ1, TNFα and LIF expressions showed increased level in the untreated patients with de novo AML, the TGFβ1 level among which is associated with the prognosis of patients.
Adolescent ; Adult ; Aged ; Case-Control Studies ; Female ; Hematopoietic System ; metabolism ; Humans ; Karyotyping ; Leukemia Inhibitory Factor ; blood ; Leukemia, Myeloid, Acute ; blood ; diagnosis ; genetics ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Transforming Growth Factor beta1 ; blood ; Tumor Necrosis Factor-alpha ; blood ; Young Adult