The clinical management of T-ALL/LBL remains challenging due to its genetic heterogeneity and variable outcomes. Integrating comprehensive genetic profiling, including CNAs and NGS mutations, is crucial for refining risk stratification.We retrospectively analyzed 66 adult T-ALL/LBL patients diagnosed between 2017 and 2021. Genetic profiling was performed using MLPA to detect key CNAs and NGS gene mutations. Clinical characteristics, OS and EFS were assessed.NGS mutations were identified in 90.9% (60/66) of patients, with NOTCH1 (46.96%), PHF6 (25.76%), and NRAS (18.18%) being the most frequent. MLPA revealed CNAs in 68.3% (41/60) of sequenced patients, with Del10q (22.73%) and Del17q (15.15%) being the most common. Multivariate analyses identified mutations in TET2 and NRAS, along with Del17q, as independent prognostic factors. A novel prognostic model was constructed by assigning one point for each of these risk factors, stratifying patients into low- (0 points), intermediate- (1 point), and high-risk (≥ 2 points) groups. This model significantly discriminated outcomes, with progressively worse OS and EFS across groups (p < 0.05). Subgroup analysis of 41 patients who underwent allogeneic hematopoietic stem cell transplantation confirmed that HSCT failed to improve the poor prognosis of the high-risk group defined by this model.Our study demonstrated the synergistic value of integrating MLPA and NGS for risk stratification in adult T-ALL/LBL.The developed prognostic model, based on TET2, NRAS, and del17q, effectively identifies a very-high-risk patient population with dismal outcomes even after allogeneic HSCT, facilitating early intervention and guiding novel therapeutic strategies.

We interrogated data from 180 consecutive subjects with myelodysplastic/myeloproliferative neoplasm with SF3B1 mutation and thrombocytosis (MDS/MPN-SF3B1-T) diagnosed according to the 2022 World Health Organization (WHO) classification of myeloid neoplasms to identify co-variates associated with survival. At a median follow-up of 45 months (Inter-Quartile Range [IQR], 19–73 months), median survival was 70 months (95% Confidence Interval [CI], 57, 84 months). Subjects with bone marrow ring sideroblasts < 15% had briefer median survival compared with those in whom it was ≥ 15% (41 months [32, 50 months] versus 80 months [63, 97 months]; P = 0.003). In multi-variable analyses, age ≥ 65 years (P = 0.005), haemoglobin concentration < 80 g/L (P = 0.008), platelet concentration ≥ 800×10E + 9/L (P = 0.02), bone marrow ring sideroblasts < 15% (P = 0.007), complex cytogenetics (P = 0.04) and SETBP1 mutation (P = 0.004) were independently associated with worse survival. Using these data we divided subjects into 3 risk cohorts: (1) low- (N = 77; median 96 months [72, 120 months]); (2) intermediate- (N = 70; median 56 months [32, 80 months]); and (3) high-risk (N = 7; median 17 months [15, 19 months]; All pairwise comparisons P < 0.0001). Our model, if validated, should help physicians estimate survival in persons with MDS/MPN-SF3B1-T in the 2022 WHO classification.

Objectives: This study evaluates the potential exosomal microRNAs (miRNAs) and associated molecular mechanisms during the progression of multiple myeloma (MM). Methods: Serum exosomes RNA isolation and transcriptome sequencing were assessed in MM patients and healthy individuals. The differentially expressed miRNAs (DEmiRNAs) were assessed depending on the sequencing data. miRNA-mRNA relations analysis, enrichment analysis, protein-protein interaction (PPI) network investigation, miRNAs and mRNAs and competing en-dogenous RNAs (ceRNA) network analysis were performed. Results: 665 miRNAs were identified from the raw exosome RNA sequencing data. A total of 46 DEmiRNAs and 419 miRNA-mRNA interaction relations were found in the two groups. The mRNAs in miRNA-mRNA interactions were gathered in functions of salivary gland development and pathways like PI3K (phosphoinositide 3-kinases)-Akt (protein kinase B) signaling. Finally, a ceRNA network constructed with several lncRNA-miRNA-mRNA interactions including SPACA6P-AS-miR-125b-5p-signal transducer and activator of transcription 3 (STAT3) was created. Conclusions: Salivary gland development function and PI3K-Akt signaling pathway enriched by exosome-based gene contributed to the development of MM. Moreover, exosome-derived miR-125b-5p and STAT3 in SPACA6P-AS-miR-125b-5p-STAT3 axis may potentially be used as biomarkers for the progression of MM.

T-cell large granular lymphocyte leukemias (T-LGLL) is a heterogeneous disease with special histology, immune phenotype and genetic abnormalities. The 2017 World Health Organization (WHO) Classific...

This study aimed to compare interphase fluorescence in situ hybridization (iFISH) and multiplex ligation dependent probe amplification (MLPA) for identifying genetic changes in myelodysplastic syndromes (MDS). The frequencies of cytogenetic changes in MDS patients treated at the Institute of Hematology and Blood Disease Hospital (China) in 2009 to 2018 were assessed by iFISH based on bone marrow samples. Then, the effectiveness of MLPA in detecting these anomalies was evaluated. Specimens from 287 MDS patients were assessed. A total of 36.9% (103/279) of MDS cases had chromosomal abnormalities detected by iFISH; meanwhile, 44.1% (123/279) harbored >= 1 copy-number variation (CNV) based on MLPA: +8 (n=46), -5 (n = 39), -7 (n = 27), del 20 (n = 32) and del 17 (n = 17). Overall, 0 to 4 aberrations/case were detected by MLPA, suggesting the heterogeneous and complex nature of MDS cytogenetics. There were 29 cases detected by MLPA, which were undetected by FISH or showed low signals. Sixteen of these cases had their risk classification changed due to MLPA detection, including 9 reassigned to the high-risk IPSS-R group. These findings demonstrated that MLPA is highly efficient in assessing cytogenetic anomalies, with data remarkably corroborating FISH findings (overall consistency of 97.1%). The sensitivities of MLPA in detecting +8, -5, -7, del 20 and del 17 were 92.3%, 97.1%, 100%, 100%, and 90%, respectively, with specificities of 95.8%, 97.6%, 97.7%, 97.6%, and 97%, respectively. MLPA represents a reliable approach, with greater efficiency, accuracy, and speed than iFISH in identifying cytogenetic aberrations in MDS.

Chromosomal abnormalities play an important role in classification and prognostication of myelodysplastic syndrome (MDS) patients. However, more than 50% of low-risk MDS patients harbor a normal karyotype. Recently, multiplex ligation-dependent probe amplification (MLPA) has emerged as an effective and robust method for the detection of cytogenetic aberrations in MDS patients. To characterize the subset of MDS with normal karyotype or failed chromosome banding analysis, we analyzed 144 patient samples with normal karyotype or undetectable through regular chromosome banding analysis, which were subjected to parallel comparison via fluorescence in situ hybridization (FISH) and MLPA. MLPA identifies copy number changes in 16.7% of 144 MDS patients, and we observed a significant difference in overall survival (OS) (median OS: undefined vs 27 months, p=0.0071) in patients with normal karyotype proved by MLPA versus aberrant karyotype cohort as determined by MLPA. Interestingly, patients with undetectable karyotype via regular chromosome banding indicated inferior outcome. Collectively, MDS patients with normal or undetectable karyotype via chromosome banding analysis can be further clarified by MLPA, providing more prognostic information that benefit for individualized therapy.

HLA‐DQB1*04:62 differs from HLA‐DQB1*04:01:01:01 by a single nucleotide in exon 3 (433A‐>G, N113D).

OBJECTIVE:To investigate the consistency between FCM and PCR on the detecting of MRD in TCF3-PBX1+ ALL, and to investigate the prognosis value of these 2 methods.METHODS:55 cases of paediatric TCF3-PBX1+ ALL patients from April 2008 to April 2015 were enrolled and analyzed. The FCM and PCR was used to detect the MRD in 239 bone marrow samples of 55 patients. All statistical analyses were carried out by using SPSS software version 16.RESULTS:Among the 55 children with TCF3-PBX1+ ALL, there were 30 male and 25 female. The median age was 5 (1-14) years. 20 patients relapsed during follow-up. The MRD results from PCR and FCM showed a strong correlation between both methods (K=0.774, P<0.001). There was no significant difference in 5-years DFS and OS between the patients in PCR+ and PCR- groups on day 15 or day 33. The 5 year DFS rate between the patients in FCM- and FCM+ was 63.9%±7.0% and 0; the 5 year OS rate was 66.5%±7.9% and 0. Combined with the result of FCM and PCR, at the d 33 of treatment, the 5-year DFS rate in FCM-/PCR- and single positive group was 65.4%±7.2% and 25.0%±15.3% (P<0.01).CONCLUSION:The detection result of MRD in TCF3-PBX1 detect by FCM and PCR shows better consistency. MRD positivity detected by FCM at the end of induction therapy (day 33) predicts a high risk of relapse in TCF3-PBX1 ALL patients.

Objective:To investigate the correlation of miRNA-181b (miR-181b) and prognostic factors of myelodysplastic syndrome (MDS), to predict target gene and main biological functions of miR-181b, and to evaluate the risk prediction ability of miR-181b in MDS.Methods:The samples of 131 bone marrow in MDS patients who followed the criteria of World Health Organization (WHO) classification (2016) from the Blood Diseases Hospital, Chinese Academy of Medical Sciences between January 2019 and September 2019 were collected, and the clinical data including routine blood test results, related gene test results of blood diseases were retrospectively analyzed. The expression levels of miR-181b in all bone marrow samples were detected by using quantitative real-time polymerase chain reaction (qRT-PCR). According to the international prognostic scoring system (IPSS), WHO classification-based prognostic scoring system (WPSS) and revised IPSS (IPSS-R), the patients were divided into different groups by the risk grade, and the expression differences of miR-181b in different risk groups were compared, and the correlation between the expressions of miR-181b and partial prognostic factors, including white blood cell (WBC), hemoglobin (Hb), platelet (Plt), absolute neutrophil count(ANC), myeloblast and gene mutations was analyzed. Bioinformatics online tool TargetScan was used to make target gene prediction and the potential function of miR-181b.Results:The expression levels of miR-181b was increased with the increasing risk of IPSS, WPSS and IPSS-R, and there were statistically significant differences in miR-181b expression levels of different risk groups in different scoring systems (all P < 0.01). There was a positive correlation between the expression level of miR-181b and the scores of the three prognostic scoring systems (r was 0.437, 0.368, 0.327; all P = 0.001); miR-181b expression was positively correlated with the proportion of bone marrow myeloblasts ( r = 0.450, P < 0.01) and was negatively correlated with Plt ( r = -0.199, P = 0.024). And miR-18b was not associated with WBC, Hb, ANC, and related gene mutations of blood diseases (all P > 0.05). A total of 1 363 potential target genes of miR-181b were predicted by using bioinformatics, and biological processes of these target genes were mainly enriched in transcription regulation, RNA metabolism regulation. Among them, 22 target genes were related to the hematological malignancies, including RUNX1, ASXL2, NRAS, ATM and KRAS, which have been previously confirmed to be related to MDS. The relative expression level [the median ( P25, P75)] of miR-181b in patients who had those hematological malignancies related to miR-181b target gene mutation (32 cases) was 1.33(0.63, 1.60), which was higher than that in patients without mutation (99 cases) [0.85 (0.49, 1.38)], and the difference was statistically significant ( Z = 2.285, P = 0.022). Conclusions:miR-181b has a correlation with the risk grade of prognostic scoring systems in MDS, and it may be involved in the molecular biology pathogenesis of MDS.

HLA‐C*15:144 differs from HLA‐C*15:07 (343G‐>A, G91R).

HLA-C*15:144 differs from HLA-C*15:07 (343G->A, G91R).

Objective: Analysis of the molecular characteristics of eosinophilia. Methods: Targeting sequence to 24 patients with chronic eosinophilic leukemia (CEL) with rearrangement of PDGFRA, PDGFRB, or FGFR1 and 62 patients with hyper-eosinophilic syndrome (HES). Mutation annotation and analysis of amino acid mutation using authoritative databases to speculate on possible pathogenic mutation. Results: Thirty-seven kinds of clonal variant were detected from 17 patients with CEL, no recurrent mutation site and hot spot region were found. No pathogenic mutation was detected in 19 patients with PDGFRA rearrangement, but pathogenic mutations of ASXL1, RUNX1 and NRAS were detected from 2 patients with FGFR1 rearrangement who progressed to acute myeloid leukemia and 1 patient with PDGFRB rearrangement who progressed to T lymphoblastic lymphoma, respectively. One hundred and two kinds of clonal abnormalities were detected in 49 patients with HES. The main hot spot mutation regions included: CEBPA Exon1, TET2 Exon3, ASXL1 Exon12, IDH1 Y208C, and FGFR3 L164V. CRRLF2 P224L and PDGFRB R370C point mutations were detected separately in 2 patients with HES who treated with imatinib monotherapy and achieved hematologic remission. Conclusion: The pathogenesis of CEL with PDGFRA, PDGFRB or FGFR1 rearrangement is usually single, and the progression of the disease may involve other driver mutation. A variety of genes with hot mutation regions may be involved in the pathogenesis of HES, and some mutation sites are sensitive to tyrosine kinase inhibitors.

Cytogenetic analysis provides important diagnostic and prognostic information for patients with Myelodysplastic syndromes (MDS) and plays an essential role in the International Prognostic Scoring System (IPSS) and the revised International Prognostic Scoring System (IPSS-R). Multiplex ligation-dependent probe amplification (MLPA) assay is a recently developed technique to identify targeted cytogenetic aberrations in MDS patients. In the present study, we evaluated the results obtained using an MLPA assay in 437 patients with MDS to determine the efficacy of MLPA analysis. Using R-banding karyotyping, 45% (197/437) of MDS patients had chromosomal abnormalities, whereas MLPA analysis detected that 35% (153/437) of MDS cases contained at least one copy-number variations (CNVs) .2/5 individuals (40%) with R-band karyotype failures had trisomy 8 detected using only MLPA. Clonal cytogenetic abnormalities were detected in 20/235 (8.5%) MDS patients with a normal R-band karyotype, and 12/20 (60%) of those patients were reclassified into a higher-risk IPSS-R prognostic category. When sequencing and cytogenetics were combined, the fraction of patients with MDS-related oncogenic lesions increased to 87.3% (233/267 cases). MLPA analysis determined that the median OS of patients with a normal karyotype (n=218) was 65 months compared with 27 months in cases with an aberrant karyotype (P=0.002) in 240 patients with normal or failed karyotypes by R-banding karyotyping. The high-resolution MPLA assay is an efficient and reliable method that can be used in conjunction with R-band karyotyping to detect chromosomal abnormalities in patients with suspected MDS. MLPA may also provide more accurate prognostic information.

OBJECTIVETo detect the immunoglobulin(Ig) and T cell receptor(TCR) gene rearrangement in bone marrow of non-Hodgkin's lymphoma(NHL) patients by using BIOMED-2 standardized system, and to explore the potential clinical significance of Ig/TCR gene rearrangement.METHODSDNA was extracted in bone marrow and Formalin-fixed and Paraffin-embedded(FFPE) samples of NHL patients, the Ig/TCR gene rearrangements were analyzed by using BIOMED-2 multiple primers system and multiplex PCR assay.RESULTSAmong 235 T-NHL cases, 71.9% showed TCR gene rearrangement. The positive rate of TCRγ and the TCRβ were 57.9% and 50.2%. Out of 583 B-NHL cases, 81.6% showed Ig gene rearrangement. The positive rate of IgH and the IgK were 70.7% and 69.3%. MCL patients showed 84.8% IgH rearrangement and 75.8% IgK rearrangement, as compared with FL(34.0%, 50.9%) and DLBCL(9.2%, 16.1%) patients, the difference was statistically significant (P<0.05). Out of Ig rearrangement positive B-NHL cases, 65 showed TCR gene rearrangement. None TCR rearrangement positive T-NHL cases showed Ig gene rearrangement, 25 cases(83.3%) showed Ig gene rearrangement in FFPE samples of 30 DLBCL patients, as compared to Ig rearrangement positive rate of bone marrow, the difference was statistically significant (P<0.001).CONCLUSIONBIOMED-2 standardized Ig/TCR gene rearrangement system shows assistance for lymphoma diagnosis. The PCR sequencing analysis is much more sensitive and specific and has significance for clinical diagnosis.

To the Editor: Primary myelofibrosis (PMF) is a Philadelphia-chromosome-negative myeloproliferative neoplasm (MPN) in which JAK2V617F, CALR, and MPLW515 mutations are the most frequent driver mutations. CALR mutations were associated with younger age, more severe anemia, higher WBC and platelet levels, lower Dynamic International Prognostic Scoring System (DIPSS)-plus scores, and longer survival compared to subjects with JAK2 mutations 1. These CALR mutations involve exon 9 and were somatic insertions or deletions (indels). Type-1 (L367fs*46) and Type-2 (K285fs*47) CALR mutations were the most frequent. Some recent researches suggested potential differences among both types of mutations in mouse model 2. In clinical study, Tefferi et al. 3 showed that the favorable prognostic impact of CALR mutations may operate only in persons with Type-1 or Type-1 like mutations and that Type-2 or Type-2 like mutations had comparable survival compared to JAK2 mutations. The difference between the two types of CALR mutation remained significant after adjusting for age, ASXL1 or EZH2 mutations. However, Rumi et al. 4 observed no difference between Type-1 and Type-2 CALR mutations. Therefore, the prognostic impact of the two different types of CALR mutations is conflicting. In the current study, we focus on the prognostic impact of Type-1 or Type-1 like and Type-2 or Type-2 like mutations based on the data from a single center in China. The study was approved by the Ethical Committee of the Institute of Hematology, CAMS according to the guidelines of the Declaration of Helsinki. Four hundred and two subjects with PMF had a bone marrow sample collected at diagnosis or referral, February 1990 to February 2015 at the Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences were enrolled. Histological material was re-reviewed by a blinded pathologist and diagnosis was based on World Health Organization (WHO) criteria. Last follow up was date of last contact, date of death, or October 15, 2015. The DIPSS scores were calculated. JAK2, MPL, and CALR mutations were tested as reported. Follow-up data were available for 361 subjects. Median follow-up of survivors was 36 months (range, 1–385) months. Correlations between sample groups and clinical and laboratory data were calculated with the χ2 test for qualitative variables with discrete categories and the Mann–Whitney U-test or Kruskal–Wallis analysis of variance for continuous variables. Survival distributions were estimated by the Kaplan–Meier method and were compared between subgroups using the log-rank test. The Cox proportional hazards regression model was used to assess the correlation between variables and survival. JAK2V617F mutations were detected in 189 subjects (47%), MPLW515 mutations in 13 (3.2%), and CALR mutations in 81 (20.1%). One hundred and nineteen subjects (29.6%) had no detectable mutation in JAK2, MPL, or CALR. There were 30 (37%) Type-1, 48 (59.3%) Type-2, and 3 (3.7%) Type-1 like CALR mutations. Subjects with Type-1 or Type-1 like and Type-2 CALR mutations were compared to those with JAK2 mutations (Supporting Information Table I). Subjects with CALR Type-1 or Type-1 like mutations were younger (P = 0.06), had lower hemoglobin concentrations (P = 0.012), lower WBC levels (P < 0.001). Subjects with CALR Type-2 mutations were younger (P = 0.009), had lower hemoglobin concentrations (P = 0.001), lower WBC levels (P < 0.001), less splenomegaly (P = 0.002), and higher DIPSS (P = 0.013). Ten-years prospective survival was 64% in JAK2-mutant, 93% in Type-1 or Type-1 like CALR-mutant, and 53% in Type-2 CALR-mutant patients, as shown in Fig. 1. Overall survival (OS) was significantly briefer in subjects with Type 2 (P = 0.004, HR 6.9, 95% CI 1.9–25.4) when compared with subjects with Type 1 or Type-1 like CALR mutations (Fig. 1). Compared with subjects with JAK2 mutations, OS was longer in patients with Type 1 or Type-1 like (P = 0.082, HR 0.3, 95% CI 0.1–1.2), and was briefer in those with Type-2 (P = 0.007, HR 2.3, 95% CI 1.3–4.1) CALR mutations (Fig. 1). Kaplan–Meier analysis of overall survival of PMF patients stratified according to JAK2 and CALR mutations. In mutivariate survival analyses, subjects with CALR Type-2 mutation (P = 0.046, HR 1.8, 95% CI 1.01–3.37) had significantly briefer survival compared with subjects with JAK2 mutations after adjusting for DIPSS. And those with CALR Type-1 or Type-1 like mutation were likely to have longer survival compared with subjects with JAK2 mutations (P = 0.062, HR 0.29, 95% CI 0.076–1.065). In summary, our findings support previous report in Teferri et al. study that the prognostic advantage of CALR mutation in PMF regards only patients harboring Type-1 or Type-1 like mutations, although the difference was of borderline significance for Type-1 or Type-1 like CALR mutant vs. JAK2-mutant. A relatively small number of CALR Type 1/Type 1-like mutated subjects in our study might hamper a significant difference for survival. Moreover, we found that subjects with Type-2 mutations had briefer survival compared to JAK2V617F-mutated subjects and the difference remained significant after adjusting for DIPSS. Recently, Guglielmelli et al. 5 also reported the prognostic advantage of Type-1 or Type-1 like CALR mutant and similar survival between Type-2 or Type-2 like CALR mutant and JAK2V617F-mutant in 251 patients from one Italian center. The difference in prognostic impact of Type-2 mutation between our findings and previous reports should be further validated. Tefferi et al. 6 found the significant association of EZH2 and IDH mutations with Type-2 CALR mutations. Therefore, the prognostic impact of subclonal mutations should be taken into account in PMF patients. We are using next generation sequencing in our subjects with CALR mutations to explore the different prognostic impact between Type-1 or Type-1 like and Type-2 or Type-2 like CALR mutations. Bing Li,1,2 Zefeng Xu,1,2 Yan Li,1 Robert Peter Gale,3 Zhen Song,4 Xiaofei Ai,5 Tiejun Qin,1 Yue Zhang,1,2 Peihong Zhang,6 Gang Huang,7 and Zhijian Xiao1,2* 1MDS and MPN Centre, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China; 2State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China; 3Division of Experimental Medicine, Department of Medicine, Imperial College London, Haematology Research Center, London, United Kingdom; 4Medical Service Division, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China; 5Molecular Diagnostic Laboratory, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China; 6Department of Pathology, Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China; 7Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio Additional Supporting Information may be found in the online version of this article. Supporting Information Table 1. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Background: The FIP1L1/PDGFRA (F/P) fusion gene is the most common clonal genetic abnormality of chronic eosinophilic leukemia (CEL). Tyrosine kinase inhibitors (TKI), such as imatinib, have been demonstrated to be effective therapies for F/P mutated disease. The aim of this study was to analyze the treatment response and long term prognosis in patients with F/P mutated CEL.Methods: The clinical features and treatment responses of 33 consecutive patients with F/P mutated CEL between August 2006 and October 2014 were analyzed. The 33 cases received imatinib therapy at an initial dose of 100 mg/day (30 patients) or 200 mg/day (3 patients); the maintenance dose depended on the response condition and patient willingness. Through the follow up, the molecular responses were regularly monitored.Results: With a median follow up of 64 months, 94% of the 33 patients with F/P mutated CEL achieved a complete hematologic remission (CHR), and 97% achieved a complete molecular remission (CMR) after a median of 3 (1.5-12) months. Twenty-four cases received maintenance therapy, with a median CMR duration of 43 (588) months. Imatinib therapy was discontinued in 8 cases, including 4 cases who experienced relapse, and 4 patients who maintained CHR or CMR after discontinuing therapy with a median time of 47 (2-74) months. One case exhibited primary resistance with a PDGFRA T674I mutation.Conclusions: F/P mutated CEL has an excellent long-term prognosis following imatinib therapy. A 100 mg daily dose of imatinib is sufficient to induce remission, and a single 100 mg weekly dose maintains a durable remission. A subgroup of patients may maintain a durable remission after discontinuing therapy with a CMR.

Objective: To analyze the long term outcome of patients with hypereosinophilic syndrome(HES). Methods: The clinical characteristics and efficacy of 60 newly diagnosed HES patients who received corticosteroids(CS)monotherapy were retrospectively analyzed. The survival and death causes of patients were obtained by follow- up. Results: Of all 60 HES patients, 45 were male and 15 female. The median age was 38(11-80)years old. The most frequent organ involvement of HES occurred in cutaneous(55.0%), gastrointestinal(40.0%), pulmonary(35.0%), cardiac(13.3%), vascular(10.0%)and neuromuscular system(10.0%). Single organ involvement was observed in 45.0% of the patients, two or at least three organ involvements were observed in 36.7% and 18.3%, respectively. The median daily dose of prednisone equivalent was 30(15-60)mg. The total response rate(CR plus PR)was 88.3%, and the rate was elevated to 93.3% after receiving alterative or combined treatment regimens. Thirty- eight patients with response to treatment received corticosteroid(CS)as mono(33 cases)or combined(5 cases)maintenance treatment with a median duration of 51(5-92)months; the median maintenance daily dose of prednisone equivalent was 5(1.25-40)mg. Twenty patients experienced cessation of CS. The main causes of patients' withdrawal were poor compliance after CR or ineffective treatment. The 5-year overall survival was(90.0±4.3)%, and the main cause of mortality was cardiac dysfunction. Conclusion: CS was highly effective on HES with manageable side effects. Most patients who have not obtained satisfactory effect could improve response via combination therapy. Cardiac dysfunction was the most common cause of mortality.

OBJECTIVE:To evaluate the prognostic value of JAK2, MPL and CALR mutations in Chinese patients with primary myelofibrosis (PMF). METHODS:Four hundred and two Chinese patients with PMF were retrospectively analyzed. The Kaplan-Meier method, the Log-rank test, the likelihood ratio test and the Cox proportional hazards regression model were used to evaluate the prognostic scoring system. RESULTS:This cohort of patients included 209 males and 193 females with a median age of 55 years (range: 15- 89). JAK2V617F mutations were detected in 189 subjects (47.0% ), MPLW515 mutations in 13 (3.2%) and CALR mutations in 81 (20.1%) [There were 30 (37.0%) type-1, 48 (59.3%) type-2 and 3 (3.7%) less common CALR mutations], respectively. 119 subjects (29.6%) had no detectable mutation in JAK2, MPL or CALR. Univariate analysis indicated that patients with CALR type-2 mutations or no detectable mutations had inferior survival compared to those with JAK2, MPL or CALR type- 1 or other less common CALR mutations (the median survival was 74vs 168 months, respectively [HR 2.990 (95% CI 1.935-4.619),P<0.001]. Therefore, patients were categorized into the high-risk with CALR type- 2 mutations or no detectable driver mutations and the low- risk without aforementioned mutations status. The DIPSS-Chinese molecular prognostic model was proposed by adopting mutation categories and DIPSS-Chinese risk group. The median survival of patients classified in low risk (132 subjects, 32.8% ), intermediate- 1 risk (143 subjects, 35.6%), intermediate- 2 risk (106 subjects, 26.4%) and high risk (21 subjects, 5.2%) were not reached, 156 (95% CI 117- 194), 60 (95% CI 28- 91) and 22 (95% CI 10- 33) months, respectively, and there was a statistically significant difference in overall survival among the four risk groups (P<0.001). There was significantly higher predictive power for survival according to the DIPSS-Chinese molecular prognostic model compared with the DIPSS-Chinese model (P=0.005, -2 log-likelihood ratios of 855.6 and 869.7, respectively). CONCLUSION:The impact of the CALR type- 2 mutations or no detectable driver mutation on survival was independent of current prognostic scoring systems. The DIPSS- Chinese molecular prognostic model based on the molecular features of Chinese patients was proposed and worked well for prognostic indication.

目的 探讨石蜡包埋组织标本应用BIOMED-2标准化免疫球蛋白(IG)基因重排技术检测弥漫大B细胞淋巴瘤(DLBCL)的可行性.方法 从45例石蜡包埋组织标本中提取DNA,采用BIOMED-2系统引物进行多重聚合酶链反应(PCR)扩增,并应用PCR片段分析法进行IG基因重排的克隆性分析.结果 将DNA由原浓度100～500 ng/μL稀释至50～100 ng/μL后,DNA扩增最大片段(300～400 bp)含量由10.0％提高到90.0％;45例DLBCL石蜡切片标本提取DNA进行免疫球蛋白重链基因(IGH)和免疫球蛋白kappa基因(IGK)克隆性分析,IGH+IGK阳性率达到84.4％;15例反应性淋巴组织增生标本进行IG/T淋巴细胞抗原受体(TCR)克隆性分析,未见克隆性重排.结论 稀释DNA是唯一可以既提高DNA扩增最大片段又能提高克隆性检出率的方法.在DLBCL的诊断和淋巴组织增生性疾病的鉴别诊断中,BIOMED-2标准化体系检测IGH基因克隆性重排是一种快速、可靠的方法,具有重要的临床应用价值.