[Clinical Value of Detecting ABL Kinase Domain Mutations in Patients with Chronic Myeloid Leukemia Based on High-Throughput Sequencing Technology].

OBJECTIVE: To compare the efficacy and clinical value of high-throughput sequencing (HTS) and Sanger sequencing in detecting ABL kinase domain mutations in patients with chronic myeloid leukemia (CML). METHODS: A total of 198 samples of 147 CML patients from July 2017 to March 2021 in Henan Cancer Hospital were collected and underwent high-throughput sequencing and Sanger sequencing to detect the mutations in ABL kinase domain, and the relevant clinical data were collected for comparative analysis. RESULTS: The proportion of total mutations and ≥2 mutations detected by high-throughput sequencing were significantly higher than those detected by Sanger sequencing (P =0.01; P =0.046). ≥2 mutations were detected in 22 cases, of which 5 cases (22.7%) had compound mutations. High-throughput sequencing can detect low level mutations that cannot be detected by Sanger sequencing. In 198 samples, 25 (12.6%) were low level mutations, 33 (16.7%) were high level mutations and 10 (5.1%) were mixed high and low level mutations. In the analysis of related clinical factors, the total mutation rate and the low level mutation rate in the optimal period, failure period and warning period were gradually increased (total mutation rate, P =0.016; low level mutation rate, P =0.005). The mutation rate of the samples with additional chromosomal abnormalities was also significantly increased (P =0.009). The mutation rate of patients who received first- and second-line treatment was significantly lower than that of patients who received third- or higher-line treatment (P =0.006). Analysis based on variant allele frequency (VAF) of the mutation site was helpful to visually evaluate the clonal evolution status of TKI-resistance CML cells. CONCLUSION: High-throughput sequencing is more sensitive and accurate than Sanger sequencing in mutation detection, which is helpful to accurately and visually evaluate TKI treatment response and optimize treatment strategy for CML.

[Strategy Optimization and Clinical Analyze of Multiple Nucleotide Polymorphism Analysis in the Chimerism Detection after Allogeneic Hematopoietic Stem Cell Transplantation].

AbstractObjective: To investigate the sample selection, result correction and clinical application value of multi nucleotide polymorphism chimerism detection method based on Next-generation sequencing. METHODS: The chimerism samples from November 2018 to June 2020 were collected, and Pearson correlation coefficient (r) was used to analyze the consistency of bone marrow and peripheral blood results detected by MNPseq; according to the different information integrity before transplantation, the calibration model was constructed to analyze the correction value of the micro chimerism results in each model; the clinical results were retrospectively analyzed to verify the reliability and practicability of chimerism results correction and the clinical value of MNPseq method. RESULTS: The results of bone marrow and peripheral blood chimerism detected by MNPseq method were consistent with each other and showed significant correlation (r=0.985, P<0.01). The three groups of calibration models were constructed according to different pre-transplant information. For the no donor and pre-transplant patients information group, the correction value was 1%; while for the group with pre-transplant patients and without donor information, 0.61% of the chimerism rate and 13 heterotopic points were used as the correction value; 0.26% of the chimerism rate and 21.57% of the heterotopic points were used as the correction value for the group with pre-transplantation patients and donor information. After correction, the number of the patients with incomplete chimerism decreased from 276 (74.19%) to 141 (37.91%) (P<0.01). Among 18 (18/141, 12.77%) patients with incomplete chimerism, the results of MNPseq in the patients were 25-39 days earlier than those in STR and flow MRD, and the result showed statistical significance. CONCLUSION: MNPseq method can be used to monitor chimerism with peripheral blood instead of bone marrow samples, and the results can be corrected to detect the changes of graft status in vivo in a more timely manner.

Prognostic significance of TET2 mutations in myelodysplastic syndromes: A meta-analysis.

Tet methylcytosine dioxygenase 2 (TET2) mutations occur frequently in myelodysplastic syndromes (MDS), but its prognostic impact has not been fully assessed and was controversial. Therefore, we performed a meta-analysis to evaluate the prognostic significance of TET2 mutations in MDS. PubMed, EMBASE databases and Cochrane Library were searched for studies reporting TET2 mutations and overall survival in MDS. Hazard ratios (HR) with 95% confidence interval (CI) were determined using random-effect modeling. A total of 1494 patients from nine studies were subjected to meta-analysis. The frequency of TET2 mutations was 18.34% (274/1494) in the study. MDS with TET2 mutations had similar overall survival compared to patients without the mutations (hazard ratio 1.13, 95% CI: 0.81-1.5).Our findings suggest that TET2 mutations have no prognosis impact on OS of patients with MDS. Therefore, the status of TET2 mutations cannot be served as a prognostic marker in MDS.

[Influence of different levels of irrigation and nitrogen application on the root growth and yield of spring wheat under permanent raised bed.] / æ°´æ°®äº’ä½œå¯¹å›ºå®šé“åž„ä½œæ ½åŸ¹æ˜¥å°éº¦æ ¹ç³»ç”Ÿé•¿åŠäº§é‡çš„å½±å“.

To establish an optimum combination of water and nitrogen for spring under permanent raised bed (PRB) tillage, a field investigation was carried out to assess effects of irrigation and N application on root growth, yield, irrigation water productivity and N efficiency. The experiment followed a completely randomized split-plot design, taking furrow irrigation 1200 m3·hm-2(W1), 2400 m3·hm-2(W2), 3600 m3·hm-2(W3) as main plot treatments, and N rates (0, 90, 180, 270 kg·hm-2) the sub-plot treatments. Our results showed that the root mass density (RWD) was significantly affected by irrigation and N application, the RWD of spring wheat reached a maximum at the filling stage, followed by a slow decline until maturity, while the effect of N on RWD depended on soil water conditions. The application of N2 produced the maximum RWD under W2 irrigation, the application of N1 produced the maximum RWD under W1 irrigation, and the application of N3 produced the maximum RWD under W3 irrigation. The order of irrigation regime effect on RWD of spring wheat was W2>W3>W1. The order of irrigation regime and N rate effect on RWD of spring wheat was irrigation>N>irrigation and N interaction. W2N2 treatment produced the highest RWD value. The root-to-shoot ratio (R/S) descended with the rising of irrigation water and nitrogen amount, and the combined treatment (W1N0) produced the maximum R/S. The root system was mainly distributed in the 0-40 cm soil layer, in which the RWD accounted for 85% of the total RWD in 0-80 cm soil depth. There was a significantly positive relationship between RWD in the 0-40 cm and the yield of spring wheat, RWD in the 40-60 cm had higher linear dependence on the yield of spring wheat. W2 increased the proportion of RWD in the deep soil layer (40-60 cm). The irrigation and N rate had a significant impact on biomass and grain yield of spring wheat, the biomass increased as the N rate and water amount increased, W2N2 treatment produced the highest grain yield, irrigation water productivity descended with increasing the irrigation amount, and the nitrogen agronomic efficiency descended with increasing N rate. It was concluded that the irrigation level W2(2400 m3·hm-2) and nitrogen level N2(180 kg·hm-2) could be recommended as the best combination of water and N, which promoted the root growth, improved grain yield, water and nitrogen use efficiencies of spring wheat production under PRB tillage in the experimental area.

[Effects of tillage type on soil organic carbon and its distribution in oasis irrigation area].

A long-term trial was established in 2005 in the oasis irrigation area to determine the impact on the accumulation and distribution of total organic carbon (TOC) , particulate organic carbon (POC) and soil microbial biomass carbon (SMBC) in 0-90 cm soil layer of 4 types of tillage including conventional tillage (CT), fresh raised-bed (FRB), permanent raised-bed (PRB) and zero tillage with control traffic on flat field (ZT). The results revealed that the distribution characteristics of TOC, POC and SMBC in the soil profile were similar in the four tillage treatments, and the carbon content decreased with depth, meanwhile the difference among treatments gradually decreased. PRB significantly increased the TOC, POC contents and SMBC, which presented in the order of PRB>ZT>FRB>CT in the 0-90 cm soil layer. In 0-10 cm soil layer, the TOC was increased by 11.1%-24.8% for PRB, 9.1%-18.7% for ZT and 7.8%-8.2% for FRB when compared with CT; POC was increased by 24.1%-26.5% for PRB, 17.3%-18.7% for ZT, and -8.2% to 10.8% for FRB; SMBC was increased by 20.5%-28.3% for PRB, 10.4%-15.2% for ZT and 3.5%-3.7% for FRB. TOC had a significant promotion effect on POC. PRB significantly increased the proportion of soil POC and enhanced the overall accumulation of organic carbon.

[Detection of NPM1, FLT3 and C-KIT mutations in acute myeloid leukemia and their prognostic analysis].

This study was aimed to evaluate the frequencies and prognostic significance of the nucleophosmin 1 (NPM1) mutation, the fms-like tyrosine kinase 3 (FLT3) mutation and c-KIT mutation in acute myeloid leukemia (AML) and to explore their relevance to clinical characteristics, cytogenetics and survival. Genomic DNA from 78 newly diagnosed AML from August 2010 to October 2012 was screened by PCR and sequencing or capillary electrophoresis (CE) for NPM1, FLT3 and c-KIT mutations. The results showed that the incidence of NPM1 mutation was 14.1% in AML patients and 26.7% in normal karyotype AML patients. NPM1 mutant cases were significantly associated with old age (P < 0.05), high peripheral white cell count and platelet counts (P < 0.05) and low expression of CD34 (P < 0.05), but no statistic difference was found in sex, percentage of bone marrow blasts, Hb, expression of CD117 and HLA-DR, complete remission rate, overall survival and relapse rate (P > 0.05). The prevalences of FLT3-ITD and FLT3-TKD mutations were 11.5% (9/78) and 3.8% (3/78) respectively, and no one patient has both of the two mutations. Patients with FLT3-ITD mutation had higher white blood cell counts and percentage of in bone marrow blasts (P < 0.05), and lower overall survival (P < 0.05), more relative to normal karyotype (P < 0.05), while no statistic difference was found in sex, age, platelet count, Hb level, complete remission rate and relapse rate (P > 0.05). No statistic analysis was performed due to the cases of less FLT3-TKD mutation. C-KIT mutation accounts for 7.7% (6/78). Patients with C-KIT mutation had a higher percentage in abnormal karyotype (P < 0.05), and higher relapse rate (P < 0.05), and lower overall survival, whereas no statistic difference was found in sex, age, percentage of bone marrow blasts, peripheral blood cell count, complete remission rate (P > 0.05). It is concluded that the detection of NPM1, FLT3 and C-KIT mutations may contribute to guiding treatment and evaluating prognosis of patients with AML.