中国实验血液学杂志最新文献

Objective: To investigate the role of platelet-derived zyxin in promoting tumor migration by platelets.

Methods: The gene expression profile of platelets was analyzed from cancer patients by using the GEO database. Isolated platelets from wild-type (WT) and Zyx^-/- mice were co-cultured with B16F10 cells labeled with green fluorescence to investigate the influence of zyxin deficiency on tumor cell migration, invasion, and wound healing. Optical microscopy was employed to evaluate the impact of zyxin deficiency on epithelial-mesenchymal transition (EMT) in B16F10 cells induced by platelets. Employing specific markers to label platelets, fluorescence confocal microscopy was utilized to investigate the impact of platelet-derived zyxin on the binding between tumor cells and platelets. And an aggregometer was employed to observe the influence of zyxin deficiency on tumor cell-induced platelet aggregation.

Results: Compared to platelets from healthy volunteers, zyxin was upregulated in platelets from cancer patients. Zyx^-/- mouse platelets exhibited a significant reduction in tumor cell invasion and migration, impaired wound healing, and delayed tumor cell EMT compared to WT mouse platelets. Additionally, zyxin deficiency attenuated the interaction between platelets and tumor cells, and diminished the capacity for tumor cell-induced platelet aggregation.

Conclusion: Platelet-derived zyxin deficiency diminishes platelet-tumor cell interactions and weakens the ability of tumor cell-induced platelet aggregation, ultimately suppressing tumor cell migration.

Objective: To analyze the genetic characteristics of the VWF gene c. 7332G>A nonsense mutation and explore its molecular pathogenesis.

Methods: Phenotypic diagnosis of the proband was performed using VWF:Ag, VWF:RCo, FⅧ:C and multimeric analysis. The probands were genotyped by NGS whole-exome sequencing, and the sequencing results were validated by sanger sequencing. The family members were genotyped by Sanger sequencing. The VWF gene c. 7332G>A nonsense mutant plasmid was constructed. After transfection, the function of VWF gene c. 7332G>A mutant plasmid was verified at cell level in vitro. The mRNA level was detected by qRT-PCR, and the expression level of protein was detected by Western blot, the function of multimerization was verified by the multimeric analysis.

Results: VWF:Ag and VWF:RCo were all less than 3% in the proband, and the multimeric analysis showed multimer deficiency. The proband was diagnosed as type 3 VWD. The homozygous nonsense mutation of VWF gene c.7332G>A was detected by gene sequencing. The VWF mRNA level of the mutant plasmid was decreased, and the VWF protein expression in the cell supernatant was decreased, the mutant protein was truncated and the function of VWF multimerization was impaired.

Conclusion: A homozygous mutation in exon 43 of VWF gene, c.7332G>A, was responsible for the probands type 3 VWD in the proband. The mutation caused a decrease in the relative level of VWF mRNA and protein, and impaired the function of VWF multimerization.

Objective: Non-targeted metabolomics based on ultra-performance liquid chromatography mass spectrometry (UPLC-MS) platform was used to study the metabolic characteristics of multiple myeloma (MM) patients, and explore potential new metabolic mechanisms affecting the occurrence and development of MM.

Methods: The study enrolled 42 MM patients, including 21 newly diagnosed (ND) patients and 21 relapsed patients (RP), and 21 age-sex matched healthy controls (HC) as subjects. UPLC-MS analysis platform was used to detect small molecule metabolites in serum of the subjects. Principal component analysis (PCA), orthogonal partial least-squared discriminant analysis (OPLS-DA) and 200 random permutations were used to analyze the differences of metabolic profiles among groups. Identification of differential metabolites was completed in the Human Metabolome Database (HMDB) and abnormal metabolic pathway analysis was performed using the KEGG database.

Results: The metabolic profiles of MM patients and healthy controls were significantly separated in both PCA and OPLS-DA models, while the metabolic profiles of newly diagnosed and relapsed MM patients were significantly separated only in OPLS-DA model. In ND vs HC, RP vs HC and RP vs ND cohorts, 19, 24 and 18 differential metabolites were identified respectively, mainly sphingolipids, glycerophospholipids and fatty acyl amino acids. The metabolic pathway abnormalities in newly diagnosed and relapsed MM patients were mainly manifested in sphingolipid metabolism and glycerophospholipid metabolism compared to healthy controls. Compared with newly diagnosed MM patients, relapsed MM patients were mainly manifested in sphingolipid metabolism and ether-lipid metabolism.

Conclusion: The metabolic profiles of MM patients are significantly different from those of healthy people. Relapsed MM patients and newly diagnosed MM patients have similar metabolic profiles, but there are still some differences. Glycerophospholipid metabolism, sphingolipid meta- bolism and ether-lipid metabolism may play important biological roles in the occurrence and development of MM.

Objective: To observe the efficacy and safety of polatuzumab vedotin (pola) combined with chemotherapy in the treatment of relapsed and refractory diffuse large B-cell lymphoma (R/R DLBCL).

Methods: A total of 23 patients with R/R DLBCL treated at the First Affiliated Hospital of Zhejiang University and its Liangzhu Branch from April 2023 to March 2024 were retrospectively collected. All patients were treated with pola combined with chemotherapy regimens such as BR, R-GDP, R-CHOP, or other regimens.

Results: All 23 patients were evaluable for efficacy, with 10 achieving complete response (CR), 7 partial response (PR), 3 stable disease (SD), and 3 progressive disease (PD). The most common adverse events included myelosuppression, fever, and pulmonary infection. No severe adverse events resulted in drug withdrawal.

Conclusion: Pola combined with chemotherapy demonstrates promising efficacy and a favorable safety profile in the treatment of R/R DLBCL.

Objective: To perform RHD gene detection on a blood sample with serological weak D phenotype.

Methods: A specimen received by the People's Hospital of Zhijin County was serologically identified by the microcolumn gel method and saline method. RHD gene detection was conducted by the PCR-SSP method, and the full sequence determination of the 10 exons amplified was performed. The sequencing results were compared with the ISBT database to determine the genotype. Bioinformatics tool was used to predict the functional damage of mutant proteins, and Alphafold-3 was used for tertiary structural modeling of wild-type and mutant RhD proteins, and the structures of the two proteins were compared and analyzed to explore the reasons why mutations lead to weak serological manifestations.

Results: The patient's genotype was identified as RHD*DV.5/RHD*01N.01 heterozygote, with the complete deletion of RHD genes on one chromosome, unable to express the D antigen. On the other chromosome, a G>A mutation occurred at the 697th base of the 5th exon, resulting in a partial D phenotype. This mutation causes internal hydrogen bond changes at the 233 position of RhD protein, resulting in a change in the conformation of the protein, affecting binding to the corresponding antibody.

Conclusion: The patient is a heterozygous mutant individual with RHD*DV.5/RHD*01N.01, exhibiting a partial D phenotype serologically. This variation is extremely rare and has been scarcely reported globally.

Objective: To investigate the effect of infusing different doses of graft cells on viral infections and survival after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

Methods: Clinical data of 99 patients with hematological diseases who underwent allo-HSCT in Shanxi Bethune Hospital were retrospectively analysed. The proportion of mononuclear cells (MNC) was counted microscopically, the proportions and absolute counts of CD34⁺ cells and lymphocyte subsets (including CD3⁺T, CD4⁺T, CD8⁺T, CD4⁺CD8⁺T, B, and NK cells) were detected by flow cytometry, and the infusion dose of each cell types was calculated. The patients were divided into high-dose group and low-dose group based on the median value of the infused cell dose. The effects of different doses of various graft cells on post-transplant viral infections (CMV, EBV, BKV) and survival were analyzed.

Results: The low-dose MNC (< 7.97×10⁸/kg) and high-dose CD4⁺CD8⁺T cell (≥3.02×10⁶/kg) groups had a higher Epstein-Barr virus (EBV) infection rate (P =0.031; P =0.020). The high-dose CD34⁺ cell group and low-dose CD3⁺ T, CD8⁺ T and natural killer (NK) cell groups showed a trend toward higher EBV infection rates, but the differences were not statitsically significant (P >0.05). There was no significant difference in the rates of EBV infection between the high-dose and low-dose groups of CD4⁺T and B cells (P >0.05). Multivariate analysis revealed that the doses of MNC and CD4⁺CD8⁺ T cells in the graft were independent factors influencing EBV infection (P =0.023; P =0.016). The doses of cells in the graft showed no significant impact on CMV and BKV infection, or patient survival (P >0.05).

Conclusion: Infusing a lower dose of MNC and a higher dose of CD4⁺CD8⁺T cells increases the risk of EBV infection, but does not affect the survival outcome.

Objective: To investigate the role of nucleolin (NCL) in acute myeloid leukemia (AML) Kasumi-1 cells and its underlying mechanism.

Methods: The Kasumi-1 cells were infected with lentivirus carrying shRNA to downregulate NCL expression. Cell proliferation was detected by CCK-8 assay, and cell apoptosis and cell cycle were determined by flow cytometry. Transcriptome next-generation sequencing (NGS) was performed to predict associated signaling pathways, the expression levels of related genes were measured by RT-PCR.

Results: Down-regulation of NCL expression significantly inhibited the proliferation of Kasumi-1 cells (P <0.01) and markedly increased the apoptosis rate (P <0.001). Cell cycle analysis showed significant changes in the distribution of cells in the G₁ and S phases after NCL knockdown (P <0.05), while no significant difference was observed in the G₂ phase (P >0.05). Transcriptome sequencing analysis demonstrated that differentially expressed genes in Kasumi-1 cells with low expression of NCL were primarily enriched in key signaling pathways, including ribosome, spliceosome, RNA transport, cell cycle, and amino acid biosynthesis. qPCR validation showed that the expression of BAX, CASP3, CYCS, PMAIP1, TP53 , and CDKN1A was significantly upregulated after NCL downregulation (P <0.05), with CDKN1A exhibiting the most pronounced difference.

Conclusion: NCL plays a critical role in regulating the proliferation, apoptosis, and cell cycle progression of Kasumi-1 cells. The mechanism likely involves suppressing cell cycle progression through activation of the TP53-CDKN1A pathway and promoting apoptosis by upregulating apoptosis-related genes.

β-Thalassemia is an autosomal recessive genetic disorder caused by defects in the synthesis of the β-globin chains. Due to ineffective erythropoiesis and premature destruction of red blood cells, patients suffer from anemia, iron overload, organ damage, and impaired immune system. The impairment of the immune system is mainly due to the increase in the levels of reactive oxygen species (ROS) caused by iron overload, which induces DNA oxidation and leads to DNA damage. The treatment strategies for β-thalassemia mainly include gene therapy and allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, iron overload in patients cannot be eliminated promptly after gene therapy and transplantation. Therefore, even if allo-HSCT is performed, the patient's hematopoietic function may still be impaired. Iron chelators and antioxidants have been proven to effectively intervene in the immune damage caused by iron overload. This article aims to review the research progress on the effects of iron overload on the immune system in patients with β-thalassemia, and provides relevant treatment recommendations for immune recovery.

Objective: To investigate the role of extracellular vesicles (EVs) derived from placental tissue and placental mesenchymal stem cells in supporting the growth and function of adult hematopoietic stem and progenitor cells (HSPCs), so as to optimize their culture system.

Methods: EVs were isolated from mouse placental tissue (PL-EV) and placental mesenchymal stem cells (PL-MSC-EV). These EVs were co-cultured with 3 000 adult bone marrow LKS⁺ (lineage^- c-Kit⁺ Sca-1⁺ ) cells for 72 hours at concentrations of 0, 1, 10, 50, 100, and 200 μg/ml. The proportion and absolute count of LKS⁺ cells after co-culture were analyzed by flow cytometry, while their self-renewal and multi-lineage differentiation potential were evaluated using colony-forming unit (CFU) assays.

Results: Compared to the blank control group, the proportion of LKS⁺ cells were significantly increased in PL-EV groups at concentrations ≥10 μg/ml after 72 hours of co-culture. Notably, LKS⁺ cells co-cultured at the concentration of 10 μg/ml exhibited the highest absolute count (899±171) and the highest proportion of LT-HSCs (LKS⁺ CD135^- CD34^-) (0.67%±0.07%). In the PL-MSC-EV co-culture system, the absolute count of LKS⁺ cells peaked at the concentration of 1 μg/ml (1011±99 cells), though the proportion of LT-HSCs was relatively low (0.15%±0.05%). The comparison between these two culture systems revealed that PL-EV at 10 μg/ml and PL-MSC-EV at 1 μg/ml displayed the most pronounced effects on LKS⁺ cell proliferation, but with no significant difference between them. CFU assays showed that, in the PL-EV culture system, the number of LKS⁺ colony formed in 1 and 10 μg/ml groups was not significantly different compared with the blank control group. In contrast, in the PL-MSC-EV system, the highest LKS⁺ colony-forming capacity was observed when co-cultured with 1 μg/ml PL-MSC-EV, while a significant reduction was noted at concentrations above 10 μg/ml.

Conclusion: PL-EV and PL-MSC-EV effectively support the growth and function of HSPCs. And PL-MSC-EV exhibits a superior efficacy in preserving the stemness of LKS⁺ cells, thus suggesting its potential for optimizing culture systems of HSPCs.