Zhonghua xue ye xue za zhi = Zhonghua xueyexue zazhi最新文献

Objective: This study aimed to develop a methodology for detecting FMS-like tyrosine kinase 3 (FLT3) gene internal tandem duplication (ITD) mutation burden using polymerase chain reaction-next-generation sequencing (PCR-NGS). The prognostic value of FLT3-ITD mutation burden detected with PCR-NGS in patients with acute leukemia (AL) before allogeneic hematopoietic cell transplantation (allo-HSCT) is investigated. Methods: This retrospective study developed a methodology for detecting FLT3-ITD mutational burden in bone marrow samples isolated from 65 patients with AL with FLT3-ITD mutations who received allo-HSCT from January 2021 to June 2024 at Ruijin Hospital. PCR-NGS was used for data analysis, and results were compared with conventional quantitative PCR (qPCR) . Results: The PCR-NGS assay demonstrated robust performance, with a sensitivity of 10(-6). Pretransplant complete remission with FLT3-ITD negativity via qPCR was achieved in 58 patients, comprising 25 with FLT3-ITD positivity in PCR-NGS [median VAF:0.629% (0.004% -26.350% ) ]. The 2-year probability of relapses and event-free survival (EFS) were 11.2% and 86.2% for patients with FLT3-ITD VAF of <0.1% and 30.6% and 64.5% for those having FLT3-ITD VAF of ≥0.1%, respectively (relapse: HR=3.159, 95% CI: 0.950-10.510, P=0.048; EFS: HR=2.846, 95% CI: 0.953-8.500, P=0.050). Two-year probability of relapses and EFS were 12.5% and 84.4% for patients with both negative PCR-NGS and qPCR, 26.2% and 73.8% for those with positive PCR-NGS and negative qPCR, and 28.6% and 57.1% for patients with both positive PCR-NGS and qPCR (relapse: HR=2.892, 95% CI: 1.122-7.451, P=0.0321; EFS: HR=1.784, 95% CI: 0.880-3.615, P=0.248), respectively, before allo-HSCT. Maintenance therapy with FLT3 inhibitors is a protective factor for reduced relapse rate and better overall survival rate after allo-HSCT. Conclusion: A highly sensitive PCR-NGS assay for FLT3-ITD mutational burden has been developed. Compared with qPCR, PCR-NGS demonstrates superior sensitivity and enables more accurate prediction of posttransplant outcomes in patients with AL undergoing allo-HSCT.

Remission rate of newly diagnosed B-cell acute lymphoblastic leukemia (B-ALL) based on chemotherapy was high; however, recurrence and refractory diseases still affect long-term survival, especially in adult patients. Chimeric antigen receptor T-cell (CAR-T cell) therapy is an important salvage treatment for patients with relapsed/refractory B-ALL, and it significantly improves the response rate, response quality, and survival of such patients. The mechanism and adverse reactions of CAR-T cellular therapy differed from those of traditional chemotherapy, molecular targeted drugs, antibody drugs, etc. Further, it has unique and, in some cases, more serious adverse reactions, despite its relatively high remission rate. Strengthening the full-process management of CAR-T cellular therapy helps achieve higher efficacy with better safety. The article takes one patient with relapsed/refractory B-ALL treated with CD19 CAR-T at our center as an example, and introduces the full-process management strategy of CAR-T cellular therapy, including patient selection, bridging therapy, lymphodepletion treatment, and adverse reaction management.

Objective: To evaluate the efficacy and long-term outcomes of fludarabine, cyclophosphamide, and rituximab (FCR) in treatment-naïve patients with chronic lymphocytic leukemia/small lymphocytic lymphoma (CLL/SLL) . Methods: Clinical data from 68 CLL/SLL patients treated with FCR at Jiangsu Province Hospital (August 2008-May 2021) were retrospectively analyzed to assess efficacy, safety, and survival outcomes. Results: Among 68 patients [46 males, 22 females; median age 55 (47, 60) years], 13.1% (8/61) had a complex karyotype, 32.3% (20/62) had immunoglobulin heavy variable region mutated (IGHV-M) type, 6.6% (4/61) had del (17p), and 14.8% (8/54) had del (11q). Patients received a median of 6 (4, 6) FCR cycles. The overall response rate was 88.2% (60/68), including 47.0% (32/68) complete remissions. Over a median follow-up of 82 (59, 98) months, 66.2% (45/68) experienced disease progression. Median progression-free survival was 56 (21, 123) months, while median overall survival was not reached. The 5- and 10-year PFS rates were 42.6% (95% CI: 31.9-56.8% ) and 28.7% (95% CI: 19.0-43.4% ), respectively. Poor PFS was associated with del (17p) (HR=5.04, 95% CI: 1.72-14.74, P=0.003), del (11q) (HR=5.27, 95% CI: 2.11-13.15, P<0.001), IGHV unmutated (IGHV-UM) (HR=4.11, 95% CI: 1.72-9.79, P=0.001), complex karyotype (CK) (HR=3.53, 95% CI: 1.58-7.85, P=0.002), β(2)-microglobulin >3.5 mg/L (HR=2.87, 95% CI: 1.37-6.01, P=0.005). In multivariate analysis, IGHV-UM remained an independent predictor of PFS (HR=8.63, 95% CI: 1.09-68.40, P=0.042). Sixteen patients with IGHV-M and lacking del (17p) or CK had a median PFS of 123 (58,123) months and a 5-year PFS rate of 70.7% (95% CI: 49.7-99.1% ), reaching a plateau after 5 years with no recurrences by 10 years. Common grade 3-4 adverse events included hematologic toxicity (44.1%, 30/68), infection (36.7%, 25/68), and liver dysfunction (4.4%, 3/68). Among 25 patients receiving single-agent BTK inhibitors after FCR progression, median follow-up was 45 (26, 64) months; 36% (9/25) experienced disease progression, with a median PFS time of 55 (27, 55) months. Conclusion: First-line FCR provides durable long-term benefits for patients with IGHV-M CLL without del (17p) or CK.

Establishing hematology care units (HCU) is essential for strengthening hematology-specific critical care capacity. This article outlines two HCU operational models in China-the ICU model and the pre-ICU model-and discusses practical insights from their implementation. It recommends establishing HCU within hematology wards, equipped for respiratory and hemodynamic monitoring and blood purification, and led by hematologists trained in critical care or by teams collaborating efficiently with general ICU through standardized workflows. For units not yet meeting these standards, establishing a resuscitation room and strengthening critical care training are advised. HCU primarily admit patients with severe or life-threatening bleeding due to hematologic diseases, severe infections or septic shock, early complications of newly diagnosed acute myeloid leukemia or acute promyelocytic leukemia (AML/APL), treatment-related complications, rapidly progressive or highly lethal conditions, and those requiring organ support or blood purification. Emphasis is placed on quantitative assessment of disease severity and timely transfer to the HCU, using tools such as the National Early Warning Score, quick Sequential Organ Failure Assessment (qSOFA), SOFA/ΔSOFA, and the Multinational Association of Supportive Care in Cancer risk index for patient stratification. Early implementation of sepsis bundles is emphasized. For organ support, high-flow nasal cannula oxygen therapy is prioritized to improve oxygenation. Continuous renal replacement therapy is performed when there are indications such as renal failure with oliguria or anuria, congestive heart failure, or drug overdose, etc. Treatment of the underlying hematologic disease follows a "three-tier stratification" and "watch-and-adjust" strategy, while respecting patient preferences. We encourage eligible hematology centers to actively establish HCU, explore new care models, and strengthen hematology-specific critical care capacity.

Objective: To assess the efficacy and safety of azacitidine combined with lenalidomide in MDS patients and explore potential mechanisms of therapeutic response. Methods: Sixteen MDS patients with TP53 mutations received azacitidine plus lenalidomide at ZhongDa Hospital, Southeast University (January 2021-June 2025). Efficacy and safety were assessed, and TP53 mutation status was correlated with treatment response. Whole-transcriptome sequencing and bioinformatics were used to explore molecular biomarkers associated with therapeutic efficacy. Results: Sixteen patients (median age 69.5 years, range 52-82; 8 males, 8 females) were enrolled. According to the Molecular International Prognostic Scoring System (IPSS-M), 1, 2, and 13 patients were classified as median low, high, and very high risk, respectively. Among 16 TP53-mutated patients, 11 had biallelic mutations and 5 had monoallelic mutations. Overall response rate was 56.3% (9/16), composite complete remission rate (CRc) was 31.3% (5/16), and hematology improvement rate was 25% (4/16). Among TP53-mutated patients, the response rate was 56.3% (9/16), with variant allele frequency dropping from 65.6% to 16.5% in responders (P=0.017). In patients with TP53 mutations and complex karyotype, response rate was 53.8% (7/13), with 57.1% (4/7) showing disappearance of CK post-treatment. The most common grade 3-4 nonhematologic adverse events were infections (9/16, 56.3% ), including pneumonia (4/16, 25.0% ), gastrointestinal infections (3/16, 18.8% ), perianal infections (1/16, 6.3% ) and sepsis (1/16, 6.3% ). High CBX8 expression may be linked to treatment response. Conclusion: Azacitidine plus lenalidomide is an effective and safe therapy for MDS, including patients with TP53 mutations and complex karyotypes. Treatment markedly reduces TP53 variant allele frequency in responders, and high CBX8 expression may predict therapeutic response.

Objective: To investigate the role and molecular mechanisms of transcription factor EB (TFEB) and its target genes in the treatment of multiple myeloma (MM) with bortezomib. Methods: TFEB target genes were predicted using the GTRD database (http://gtrd.biouml.org/), identifying Ptch1 gene for further study. Expression changes of Ptch1 in RPMI8226 and U266 MM cell lines after bortezomib treatment were assessed by real time fluorogenic quantitative PCR (RT-qPCR) and Western blot. RPMI8226 and U266 cell lines were transfected with siRNA-TFEB, and mRNA and protein levels of key factors (Ptch1, Gli1) in the Ptch1/Hedgehog signaling pathway were measured by RT-qPCR and Western blot. Furthermore, Ptch1 was overexpressed in MM cell lines via lentiviral transduction. Autophagy was evaluated by acridine orange staining, and protein levels of LC3B, Beclin-1, and Lamp-1 were measured by Western blot. Lysosomal quantity changes were assessed by lysosomal fluorescent probes. Results: Bortezomib (6.0×10(-6) mmol/L, 24 h) significantly reduced Ptch1 mRNA and protein levels in both cell lines compared with blank control group (all P<0.05). siRNA-TFEB transfection reversed bortezomib's inhibition of Hedgehog pathway key factors Ptch1 and Gli. Ptch1 overexpression in bortezomib-treated RPMI8226 and U266 cells significantly reduced the relative expression of autophagy-related proteins LC3B, Beclin-1, and Lamp-1 (all P=0.001). Acridine orange staining showed fewer acidic vesicular organelles within two cell lines (all P=0.001). The relative fluorescence expressions of lysosomal probes reflecting the number of lysosomes were also decreased (P values of RPMI8226 and U266 cell lines were 0.001 and 0.007, respectively) . Conclusion: The knockdown of TFEB can specifically promote the expression of the Ptch1/Hedgehog signaling pathway, thereby reducing bortezomib-induced autophagy in MM cells and reversing the inhibitory effect of bortezomib on the proliferation of MM cell lines.

Objective: To evaluate the diagnostic value of intestinal tissue metagenomic next-generation sequencing (mNGS) in severe diarrhea following haploidentical allogeneic hematopoietic stem cell transplantation (allo-HSCT) . Methods: Sixteen patients who developed severe diarrhea or hematochezia after haploidentical allo-HSCT at the First Affiliated Hospital of Fujian Medical University (June 2023-August 2024) were enrolled. All underwent gastrointestinal endoscopy and mNGS for microbial detection. Clinical, endoscopic, pathological, and microbiological data were analyzed to evaluate the diagnostic value of mNGS and treatment outcomes following targeted therapy. Results: The study included 16 patients (12 males, 4 females; median age 32.5 years, range 3-60 years). Diarrhea occurred a median of 3.93 months post-transplant (range 1.63-10.40 months). Stool cultures were negative except for one case with Candida. One patient tested positive for Clostridium difficile antigen. Endoscopy revealed mucosal congestion, edema, erosion, and bleeding, with focal inflammation on pathology. mNGS detected pathogens in 87.5% (14/16) of cases, including mixed infections in 78.5% (11/14). Common pathogens were Klebsiella pneumoniae, Enterococcus faecium, Escherichia coli, Rhizopus microsporus, EBV, and CMV. Targeted treatment adjustments led to symptom improvement in 87.5% of patients. Conclusion: Allo-HSCT patients are prone to infectious diarrhea due to immunosuppression. Molecular analysis of endoscopic biopsy tissues using mNGS can accurately identify pathogens, guide targeted therapy, and improve clinical outcomes.

Objective: Dehydrated hereditary stomatocytosis (DHS), a rare condition caused by pathogenic PIEZO1 or KCNN4 mutations, remains under-recognized. This study aimed to improve the diagnosis and management of DHS in China by retrospectively analyzing clinical and genetic characteristics. Methods: Patients diagnosed with DHS at Peking Union Medical College Hospital between 2018 and 2024 were included. All had suspected congenital hemolytic anemia and were confirmed by next-generation sequencing to harbor pathogenic PIEZO1 or KCNN4 mutations. Clinical features, genotypes, iron overload, and treatment outcomes were collected and analyzed. Results: Twenty-three DHS patients were identified, including 21 with PIEZO1 and 2 with KCNN4 mutations. Common manifestations were splenomegaly, jaundice, cholelithiasis, and secondary iron overload. Mild and moderate anemia occurred in 8.7% and 17.4% of patients, respectively, and stomatocytes were present in only 17.4%. The median time from symptom onset to diagnosis was 11 years. The predominant PIEZO1 variants were c.7367G > A and c.6328C > T, with nine novel mutations identified. Both KCNN4 mutations involved c.1055G > A. No clear genotype-phenotype correlation was observed. Secondary hepatic iron overload was found in 21.7% of patients. Conclusion: DHS is often underdiagnosed or misdiagnosed. Genetic testing enables accurate identification, and regular monitoring for secondary hepatic iron overload is crucial for long-term management.

Objective: To investigate the clinical, laboratory, and prognostic features of myelodysplastic neoplasm (MDS) patients harboring acute myeloid leukemia (AML) -like mutations. Methods: We retrospectively analyzed clinical, molecular, and outcome data from 1 464 adults with primary MDS diagnosed at the Institute of Hematology and Blood Diseases Hospital from August 2016 to June 2024. Results: AML-like mutations were detected in 64 patients (4.4% ). Compared with patients without AML-like mutations, those with AML-like mutations were younger [median 50 (IQR 39-60) vs 56 (45, 65) years; P=0.001], more often female (51.6% vs 35.4% ; P=0.009), had higher bone marrow blast percentage [6.5% (3.0%, 10.5% ) vs 2.5% (1.0%, 7.0% ) ; P<0.001], a higher rate of normal karyotype (75.0% vs 48.1% ; P<0.001), and lower hemoglobin levels [73 (67, 82) g/L vs 80 (66, 98) g/L; P=0.006]. The AML-like group had a higher number of gene mutations than the non-AML-like group [3 (IQR 2-4) vs 2 (1, 3) ; P<0.001). It was enriched for mutations in NPM1, DNMT3A, WT1, PTPN11, NRAS, BCOR, FLT3, CEBPA, and MYC (all P<0.05) and had lower rates of U2AF1, ASXL1, and TP53 mutations (all P<0.05). Overall survival (OS) did not differ between groups (P=0.730) ; however, the AML-like group had significantly shorter leukemia-free survival (LFS) [19 months (95% CI: 13-25) vs 46 months (95% CI: 38-54) ; P=0.012] and a higher 2-year cumulative incidence of AML transformation [ (41.7±9.1) % vs (10.4±1.1) % ; P<0.001]. Within the AML-like group, OS, LFS, and cumulative incidence of AML transformation did not differ between patients with low blasts and those with excess blasts (IB). Multivariable Cox regression identified age ≥60 years and PTPN11 mutations as independent adverse prognostic factors for OS, while DNMT3A, PTPN11, and FLT3 mutations independently predicted leukemic transformation. Conclusions: MDS patients harboring AML-like mutations exhibit distinct clinical and molecular features and a higher risk of progression to AML.

Objective: To evaluate the efficacy and safety of the CLAE (cladribine + cytarabine + etoposide) regimen in refractory/relapsed T cell acute lymphoblastic leukemia/lymphoma (R/R T-ALL/LBL) . Methods: Patients with R/R T-ALL/LBL received the CLAE regimen in a prospective, multicenter, single-arm clinical study or compassionate use. From March 2019 to August 2024, data from 25 patients (18 in the study across five centers and 7 receiving compassionate treatment in Peking University Third Hospital) were collected. Outcomes included objective response rate, complete response (CR) rate, partial response (PR) rate after 1-2 cycles, bridging to allo-HSCT, progression-free survival (PFS), overall survival (OS), and treatment-related adverse effects. Results: Median age was 29 years (range, 13-63) ; 17 were male. Among the 24 evaluable patients, CR rate was 33.3% overall and 41.2% among enrolled patients. Median OS and PFS time were 199 (46-1 310) and 49 (28-1 310) days, respectively. Cumulative OS rate at 6 months, 1 year, and 2 years was (52.1±10.2) %, (29.7±9.3) %, and (27.1±9.1) %, respectively; cumulative PFS rate was (32.6±9.6) %, (24.9±8.9) %, and (23.8±8.7) %, respectively. Among patients achieving CR or PR (8 cases), median OS and PFS were not reached. Cumulative OS rate at 6 months, 1 year, and 2 years was (86.8±12.0) %, (78.3±14.6) %, and (72.9±15.7) %, respectively, and the cumulative PFS rate was (86.4±12.1) %, (74.8±15.3) %, and (72.9±15.7) %, respectively. Adverse events were mainly hematologic; no treatment-related mortality occurred. Seven patients achieving CR were bridged to allo-HSCT, with 5 remaining in continuous remission. Conclusion: The CLAE regimen is safe and effective for R/R T-ALL/LBL, facilitating CR as a bridge to allo-HSCT and potentially improving patient prognosis.