中国实验血液学杂志最新文献

Objective: To construct a simple score scale for donation-related vasovagal reaction (DRVR) based on Logistic regression model and evaluate its application value.

Methods: A retrospective study was conducted on 5000 blood donors from Zhangjiakou Central Blood Station from January 2018 to January 2023. The baseline data of the blood donors were collected. Logistic regression analysis was performed on them to construct a DRVR prediction model. A DRVR simple score scale was built based on the odds ratios. The score intervals were divided into low-risk, medium risk, and high-risk categories, the application value of the scale was evaluated based on the prediction efficacy of Logistic regression model.

Results: The incidence of DRVR among the blood donors was 4.12%. The Logistic regression model identified eight independent risk factors for DRVR, including advanced age, female gender, low body mass index, first blood donation, hypotension, low hemoglobin level, long blood collection time, and high blood donation volume (all P < 0.05). A simple DRVR score scale was constructed based on the odds ratio values. The scale scores were divided into three intervals: low-risk (total score <6), moderate risk (total score 6-11), and high-risk (total score >11,≤17). The area under the curve(AUC) for predicting DRVR in low-risk, medium risk, and high-risk blood donors based on Logistic regression model were 0.784 (95% CI : 0.742-0.844), 0.806 (95% CI : 0.752-0.883), and 0.842 (95% CI : 0.761-0.925), respectively. The consistency indices were 0.794 (95% CI : 0.705-0.828), 0.800 (95% CI : 0.745-0.852), and 0.839 (95% CI : 0.782-0.917), respectively. The calibration curves did not deviate from the ideal fit. The results of external validation showed that the receiver operating characteristic curve fittings were relatively ideal (all P >0.05), and there were no statistically significant difference in AUC (all P >0.05).

Conclusion: The DRVR simple score scale has a relatively ideal ability to distinguish the blood donors and can provide some reference for quickly narrowing the scope of high-risk groups. DRVR simple score scale has certain application value, but still needs to be continuously improved and optimized.

Objective: To explore the difference of type 2 innate lymphoid cell (ILC2), IL-9 and related cytokines between chronic lymphocytic leukemia (CLL) patients and normal individuals, as well as the correlation between ILC2 and IL-9 and other cytokines in CLL patients.

Methods: Flow cytometry was used to detect the expression of ILC2 and regulatory T cells (Tregs) in peripheral blood of 26 CLL patients at initial diagnosis and 10 healthy controls. RT-qPCR was used to detected IFN-γ, TGF-β, IL-9 and IL-10 mRNA in peripheral blood mononuclear cell (PBMC). ELISA was used to detect serum IFN-γ, TNF-α, TGF-β, IL-9, IL-10 and IL-21. ILC2 and IL-9 were observed in the cervical lymph node of 12 CLL patients at initial diagnosis and 12 patients with reactive lymphoid hyperplasias by multiplex indirect immunofluorescence staining. Spearman test was used to analyze the correlation between peripheral blood ILC2 and IL-9, IL-9 and IL-21, IFN-γ mRNA and IL-10 mRNA in CLL patients. Pearson test was used to analyze the correlation between TNF-α and TGF-β in CLL patients.

Results: Compared with control group, the proportions of ILC2 and Tregs were significantly increased in CLL group (both P < 0.05). The mRNA expressions of IFN-γ, IL-9, IL-10 and TGF-β in PBMCs of CLL patients were significantly increased (all P < 0.05). In CLL patients, the expressions of IFN-γ, TNF-α, TGF-β, IL-9 and IL-10 in serum were significantly increased (all P < 0.01), while IL-21 slightly increased without statistical difference (P >0.05). In CLL patients, the peripheral blood ILC2 was positively related to IL-9 (r =0.56), IL-9 was positively related to IL-21 (r =0.397), IFN-γ mRNA was positively related to IL-10 mRNA (r =0.623), and TNF-α was positively related to TGF-β (r =0.577). Compared with reactive lymphoid hyperplasias patients, the mean fluorescence intensities of GATA3 and CRTH2 representing ILC2 and IL-9 in cervical lymph nodes were significantly increased in the CLL group (all P < 0.001), and showed colocalization.

Conclusion: In CLL patients, the proportions of ILC2 and IL-9 in peripheral blood and lymph nodes increase, and ILC2 and IL-9 show colocalization in lymph nodes. There is a positive correlation between ILC2 and IL-9 in the peripheral blood of CLL patients, the ability of ILC2 to secrete IL-9 is increased, and ILC2 may affect the occurrence and development of CLL through IL-9.

Objective: To investigate the molecular characteristics of low-risk acute myeloid leukemia (AML) at recurrence, and analyze the factors affecting retreatment efficacy and prognosis.

Methods: A retrospective analysis was conducted on the clinical and laboratory data of 31 patients with newly diagnosed low-risk AML who relapsed during consolidation treatment or follow-up after treatment in our hospital from April 2017 to January 2023. Gene mutations before and after relapse were compared, retreatment efficacy following relapse was evaluated, and univariate and multivariate analyses were performed to identify factors influencing treatment efficacy and prognosis.

Results: Gene sequencing results after relapse showed that the most common newly acquired mutation was FLT3-ITD , while RAS mutation detected at initial diagnosis were predisposed to loss of expression during relapse. The median overall survival (OS) after relapse for the entire cohort was 349 (170-528) days, with non-hematopoietic stem cell transplantation (HSCT) group and HSCT group demonstrating median survival times of 210 (106-314) days and not reached, respectively (P =0.001). Multivariate analysis revealed that age ≥60 years was a significant risk factor for achieving remission after retreatment in initially diagnosed low-risk AML patients who experienced relapse (OR =18.222, 95%CI : 1.188-279.597, P =0.037). Additionally, DNMT3A mutation was identified as an independent risk factor for OS (HR=13.165, 95%CI : 2.018-85.877, P =0.007), while HSCT post-relapse demonstrated significant survival benefits (HR=0.133, 95%CI : 0.025-0.698, P =0.017) and served as an independent protective factor for OS.

Conclusion: Relapsed low-risk AML is often associated with loss of RAS and novel mutations in FLT3-ITD . Age ≥60 years and DNMT3A mutations were identified as independent adverse factors for achieving subsequent remission and post-relapse survival, respectively, while HSCT significantly improved patient outcomes.

Triple-negative myeloproliferative neoplasms (TN-MPN) are diseases characterized by absence of the three driver mutations in JAK2, CALR, or MPL, but still exhibit histological and phenotypic features sufficient to diagnose myeloproliferative neoplasms (MPN). Approximately 10% to 20% of essential thrombocythemia (ET) and 5% to 10% of primary myelofibrosis (PMF) cases are reported to be triple negative. TN-MPN may carry non-classical driver mutations at JAK2 or MPL, or other gene mutations, including somatic mutations of chromatin structure, epigenetic modifiers (TET2, IDH1/2), splicing factors (SF3B1, SRSF2, U2AF1, ZRSR2), and cytokine signaling regulators (CBL, SH2B3), etc., and there is evidence of clonal hematopoiesis. This article reviews the latest research progress in the pathogenesis, diagnosis, clinical features, prognosis and treatment of TN-MPN.

Objective: To analyze and confirm the ambiguous results of HLA-DRB1 genotyping in one case.

Methods: HLA genotyping was performed on a sample of hematopoietic stem cell donor using Illumina MiSeq-based next-generation sequencing (NGS). The ambiguous results of HLA-DRB1 locus were further analyzed and confirmed through PacBio SMRT third-generation sequencing.

Results: The Illumina MiSeq-based NGS typing results suggested the presence of a new HLA-DRB1*11 allele (DRB1*11:NEW, 12:01) in the specimen, with a mismatch of G>A located in the 40^th residue of exon 1 compared with the nearest allele DRB1*11:01:01:03. However, due to the long sequence of intron 1, this observed mutation site was so far away from the near heterozygous sites that no reads could cover this gap. Therefore, it was impossible to determine which consensus the mutation site was located in, and the NGS-based genotyping results were obtained from the random allocation by the software, which was ambiguous and unreliable. In order to confirm the results, the long-read third generation sequencing technology based on PacBio was applied to genotype the DRB1 locus. The results showed that the DRB1 typing was HLA-DRB1*11:01,12:10. E1-40A was actually located in the allele HLA-DRB1*12:XX , which was exactly matched with HLA-DRB1*12:10.

Conclusion: For some new alleles suggested by NGS, especially the ambiguous ones that are far away from other heterozygous sites, it is necessary to analyze and confirm them by other methods such as the third-generation long-read sequencing technology to obtain reliable results.

Objective: To investigate the mechanism of the synergistic anti-leukemia effect of the combination of Bcl-2 inhibitor venetoclax (VEN) and histone deacetylase (HDAC) inhibitor chidamide (CDM) in T-cell acute lymphoblastic leukemia (T-ALL).

Methods: The effect of VEN combined with CDM on the proliferation of T-ALL CEM and MOLT-4 cell lines was detected by CCK-8 assay. And the effects on the cell cycle and apoptosis were detected by flow cytometry. Cell cycle protein and apoptosis-related protein expression were detected by Western blot. The key pathways of VEN combined with CDM in T-ALL were screened through network pharmacology analysis, and verifying them in T-ALL cell lines, T-ALL patient cells and public databases.

Results: VEN combined with CDM displayed a synergistic effect on cell proliferation of CEM and MOLT-4 cells. In cell cycle experiment, VEN combined with CDM induced G₀/G₁ phase arrest in CEM and MOLT-4 cells. Western blot experiment showed that VEN combined with CDM could significantly downregulate the expression of cyclin E2 and CDK2 and upregulate the expression of p21^Waf1/Cip1. In the apoptosis experiment, VEN combined with CDM could significantly induce the apoptosis of CEM and MOLT-4 cells. Western blot experiment demonstrated that VEN combined with CDM promoted endogenous apoptosis by downregulating Mcl-1 and upregulating Bax and cleaved caspase-3 protein levels. Network pharmacology analysis identified 10 hub genes. KEGG enrichment analysis revealed the cell cycle, PI3K-AKT signaling pathway, and its downstream FoxO signaling pathway were significantly enriched. GO enrichment analysis revealed the G₁/S transition of mitotic cell cycle, cyclin-dependent protein kinase holoenzyme complex, and kinase activity were significantly enriched. Western blot experiment showed that VEN combined with CDM could significantly downregulate the protein level of PI3K, AKT, and p-AKT, and upregulate FoxO1 in CEM and MOLT-4 cells. In T-ALL patients, FoxO1 showed significantly lower expression compared to the normal donors, and the same result was verified in the GSE13159 and GSE26713 datasets.

Conclusion: The combination of VEN and CDM exerts synergistic anti-leukemia effects by inhibiting cellular proliferation, inducing G₀/G₁ phase arrest and promoting apoptosis through PI3K/AKT/FoxO1 axis in T-ALL.

Objective: To investigate the role of microRNA-709 (miR-709 ) in erythroid development and its correlation with multiple hematological diseases.

Methods: The expression of miR-709 in multiple tissues of mice was detected by qRT-PCR; The expression of miR-709 and other miRNAs in day-14.5 fetal liver cells from mouse embryos was detected by transcriptome microarray analysis; The expression of miR-709 in nucleated red blood cells derived from bone marrow and spleen, and in peripheral blood erythrocytes of mice was detected by magnetic bead sorting combined with qRT-PCR; The expression of miR-709 during erythroid differentiation of murine erythroleukemia (MEL) cells was analyzed by cell culture and qRT-PCR; The expression of miR-709 during erythroid lineage differentiation of mouse erythroid precursor cells derived from fetal livers was analyzed by magnetic bead sorting, flow cytometry, cell culture and qRT-PCR; The expression of miR-709 in peripheral blood of patients with different hematological diseases was measured by qRT-PCR.

Results: Among the various tissues examined in mice, miR-709 exhibited the highest expression in peripheral blood, followed by high expression levels in muscle, bone marrow, and liver; In day-14.5 fetal liver cells from mouse embryos, miR-709 was highly expressed, significantly surpassing miR-451, which was most highly expressed in mature red blood cells; In nucleated red blood cells derived from mouse bone marrow and spleen, the expression of miR-709 was higher than that of miR-451 , whereas the opposite pattern was observed in peripheral blood; During the differentiation of erythroid precursor cells derived from mouse embryonic liver, the expression level of miR-709 first increased and then decreased; During the differentiation of MEL cells, the expression of miR-709 gradually increased; Compared with the healthy controls, patients with myelodysplastic syndrome (MDS), α-thalassemia and β-thalassemia expressed lower levels of miR-709 in peripheral blood; while the expression of miR-709 in patients with infectious hemolytic anemia (IHA) was higher than that in healthy controls.

Conclusion: miR-709 is highly expressed in the early stage of erythropoiesis and exhibits dynamic changes during erythroid development, potentially playing an important role. It is also differentially expressed in different hematological diseases, which is expected to serve as a promising biomarker and therapeutic target in the clinical diagnosis and treatment of hematological diseases.

Objective: To evaluate the efficacy and safety of blinatumomab in adult patients with relapsed/refractory (R/R) or measurable residual disease (MRD) positive B-cell acute lymphoblastic leukemia (B-ALL) in the real world.

Methods: The clinical data of 30 B-ALL patients received at least 1 course of blinatumomab therapy in the Chinese PLA General Hospital from January 1^st, 2021 to December 31^st, 2023 were retrospectively analyzed, including pre-treatment baseline clinical feature, post-treatment complete response (CR), CR with partial hematologic recovery (CRh), CR with incomplete hematologic recovery (CRi), complete MRD response rate, MRD response rate (MRD< 10^-4), overall survival (OS), and disease-free survival (DFS), as well as drug-related adverse reactions.

Results: Among 5 patients who were not assessed 4 were MRD negative and 1 did not receive bone marrow biopsy. In the R/R B-ALL group (13 cases), 11 patients achieved CR/CRh/CRi and 10 patients achieved complete MRD response. In MRD⁺ group (12 cases), 9 patients achieved overall MRD response and 7 patients achieved complete MRD response. The median follow-up time was 8.4(95%CI : 6.3-10.4) months. The median OS was 15.5(95%CI : 0.7-30.3) months in the R/R group, while not reached in the MRD⁺ group. The median DFS of the two groups were not reached. Drug-related adverse reactions occurred in 22 patients, and pyrexia was the most common (13 cases). Grade ≥3 adverse reactions occurred in 15 patients, and neutropenia was the most common (9 cases). Cytokine release syndrome occurred in 6 patients, including 5 cases with grade 1 and 1 case with grade 3. No patients interrupted therapy or died due to drug-related adverse reactions.

Conclusion: Blinatumomab is effective in the treatment of R/R or continuous MRD⁺ B-ALL with acceptable adverse reactions.

Objective: To investigate the effects of TP53 genetic status (wild-type/mutated/null) on the drug resistance of decitabine (DAC) in myelodysplastic syndromes (MDS) and identify key resistance-associated genes.

Methods: Two myeloid cell lines with distinct TP53 status (M-07e: wild-type; SKM-1: mutated;) were treated with gradient DAC concentrations (0-10 μmol/L) for 0-72 h. Cell viability was detected by CCK-8 assay. RNA-Seq transcriptomics, and methylation profiling were integrated to analyze differentially expressed genes.

Results: Decitabine (DAC) treatment induced time- and dose-dependent inhibition of cell viability in CCK-8 assays, with SKM-1 cells exhibiting the highest resistance (IC₅₀=5 μmol/L vs M-07e=0.5 μmol/L, P < 0.01). Transcriptomic analysis revealed 662 upregulated and 452 downregulated genes in DAC-treated M-07e cells, while SKM-1 cells showed 515 upregulated and 73 downregulated genes. By proteomic profiling, 117 upregulated and 136 downregulated proteins were identified in M-07e cells, while 91 upregulated and 46 downregulated proteins were identified in SKM-1 cells following DAC exposure. Through integrated analysis of upregulated genes and proteins expression profiles, 181 candidate genes were screened out, while methylation studies identified 884 hypomethylated genes with high-sensitivity loci and CpG density. Notably, 31 genes overlapped between these datasets, and functional annotation indicated these drug-resistance-associated genes are primarily involved in positive regulation of cell differentiation, negative regulation of binding processes, and negative regulation of cellular component organization.

Conclusion: TP53 mutations drive DAC resistance via epigenetic reprogramming. Targeting these genes may improve outcomes in TP53 -mutated MDS.