中国实验血液学杂志最新文献

Objective: To explore whether Resveratrol (RSV) can inhibit the spontaneous senescence of human bone marrow-derived mesenchymal stem cells (MSC).

Methods: MSC were serially cultured to passage 13 and passage 15 to establish model groups exhibiting spontaneous senescence, respectively. MSC at passage 13 and passage 15 were treated with 5 nmol/L RSV for 48 h to establish the RSV-treated groups. SA-β-Gal staining was used to detect cell senescence. MTT assay was used to detect cell proliferation. RT-PCR method was used to detect senescenceassociated telomerase activity. Western blot was used to detect the senescence-associated protein level of the phosphorylated-mTOR.

Results: SA-β-Gal staining showed that the senescent cells of MSC in RSV-treated group was significantly less than those in the model group (RSV group compared with model group at passage 13, P < 0.05; RSV group compared with model group at passage 15, P < 0.01). The cell proliferation ability of MSC in RSV-treated group was significantly higher than those in model group, at 72 h in passage 13, there was significant difference between RSV-treated group and model group (P < 0.05). RT-PCR results showed that the hTERT mRNA expression of MSC in RSV-treated group was higher than that in model group, which was significantly different between RSV-treated group and model group at passage 13 (P < 0.05). Western blot results showed that the phosphorylated (Ser2448)-mTOR level of MSC in RSV-treated group was lower than that in model group, which was significantly different between RSV-treated group and model group at passage 13 (P < 0.05).

Conclusion: RSV can inhibit the spontaneous senescence of human MSC by mediating mTOR activity.

Objective: To investigate the expression level, clinical significance and function of circular RNAs (circRNAs) circ_0073585 in the bone marrow of patients with acute myeloid leukemia (AML).

Methods: The expression levels of circ_0073585 in bone marrow samples of 106 newly diagnosed AML patients and 38 controls were detected by real-time quantitative PCR (RQ-PCR). The differences were compared between the two groups and their clinical significance was analyzed. The diagnostic value of circ_0073585 expression for AML was evaluated by receiver operating characteristic curve(ROC). THP-1 cells with lentivirus overexpressing circ_0073585 vector and empty vector were divided into two groups: circ_0073585-THP-1 and NC-THP-1 group. CCK-8 assay and flow cytometry were used to study the effects of circ_0073585 on THP-1 cell proliferation, survival, apoptosis and drug sensitivity.

Results: Compared with the controls, the expression level of circ_0073585 in the bone marrow of AML patients was significantly reduced (P < 0.001). There was a statistically significant difference between the high and low expression groups of circ_0073585 in the white blood cell count, platelet count (P < 0.01) and chromosome risk (P < 0.05). Compared with NC-THP-1 cells, the proliferation and viability of circ_0073585-THP-1 cells were weakened (P < 0.01), the apoptosis rate was increased (P < 0.01), and the sensitivity to homoharringtonine (P < 0.05) and daunorubicin hydrochloride (P < 0.001) was increased.

Conclusion: The expression level of circ_0073585 is decreased in AML patients. Overexpression of circ_0073585 can inhibit the proliferation and viability of leukemic cells, promote apoptosis, and increase sensitivity of leukemia cells to chemotherapy drugs.

Objective: To establish the absorption and elution test for relatively quantitive obtaining anti-A and anti-B blood group IgG antibodies in the plasma of O-type RhD-positive pregnant women.

Methods: 95 cases of the O-type RhD-positive pregnant women plasma samples were randomly selected for obtaining the IgG antibodies of anti-A and anti-B blood group, with absorption test under 37 ℃ and elution test under 56 ℃, and the IgG anti-A and anti-B antibody titers of plasma and elution were determined by the microcolumn gel anti-human globulin test. The differences and correlation between the titers of IgG antibodies in the eluent and plasma were compared and analyzed.

Results: After a logarithmic transformation (Log2), there was no statistically difference between IgG antibody anti-A difference value and anti-B difference value in the eluent and plasma (P >0.05). The titer of IgG antibody in the eluent was positively correlated with the titer of IgG antibody in the plasma (r =0.914). The linear equation for IgG antibody titers fitted by a scatter plot between the eluent and plasma was Y=-3.55+0.96X.

Conclusion: The absorption and elution test can be used to obtain the anti-A and anti-B IgG antibodies in the plasma of O-type RhD-positive pregnant women, whose plasma origin IgG titer is greater than 8. Meanwhile, the acquisition of anti-A antibodies was as effective as anti-B antibodies at the same time, and the antibodies obtained are positive proportional to their respective concentrations in the plasma.

Objective: To explore the overall survival and prognostic factors of patients over 50 years old with acute myeloid leukemia (AML).

Methods: The clinical data of 222 AML patients aged over 50 years in our hospital from January 2016 and June 2021 were retrospectively analyzed. Kaplan-Meier method was used to evaluate the overall survival (OS) rate, and Cox regression model to evaluate the prognostic factors.

Results: The 1-year and 3-year OS rates of all patients were 46.8% and 28.8%, respectively. The recurrence rate of patients who achieved remission during follow-up time was 57%. Both univariate and multivariate analysis showed that advanced age, MLL family fusion gene, PHF6 gene mutation, TP53 gene mutation, intolerance to standard chemotherapy, incomplete remission, complex karyotype, +mar karyotype and inv(3) karyotype were significantly correlated with prognosis (all P <0.05). Negative fusion gene and positive AML- ETO fusion gene had no obvious survival advantage in this population. In patients with complete remission, there was no significant survival advantage in those who achieved minimal residual disease negative.

Conclusion: AML patients aged over 50 years have a poor outcome and high recurrence rate. The prognosis is affected by multiple factors and has its own characteristics.

Objective: To analyze the relationship between serum cystatin C (CysC), β₂-microglobulin (β₂-MG) and the efficacy of demethylation therapy in patients with acute myeloid leukemia (AML).

Methods: A prospective cohort study was conducted on 98 AML patients admitted to the Affiliated Hospital of Inner Mongolia Medical University from February 2019 to January 2022. All patients were treated with decitabine (DAC) + HAG regimen, 28 days as a course and treated for 3-4 courses. At the end of each course of treatment, the treatment effect of the patients was evaluated, and the patients who achieved complete remission (CR) transferred to consolidation therapy, while the patients who did not reach CR at the end of the course of treatment were considered as treatment failure. The examination items before treatment include routine blood parameters, serum CysC, and β₂-MG, and general clinical data of the patients were collected. According to the statistical results, logistic regression model was used to analyze the relationship between serum CysC, β₂-MG and the efficacy of demethylation therapy in AML patients. The ROC curves were drawn, and the predictive efficacy of serum CysC, β₂-MG on demethylation therapy in AML patients was evaluated by the area under the curve (AUC).

Results: Of the 98 AML patients enrolled in the study, 5 cases were excluded during the treatment period, and 93 cases finally completed the chemotherapy courses. Among them, 23 patients achieved CR after the initial induction chemotherapy (course 1-2), and 11 patients achieved CR after the re-induction chemotherapy (course 3-4). The success rate of demethylation therapy was 36.56 % (34/93). Compared with the patients in treatment success group, patients in treatment failure group had a higher proportion of intermediate- and adverse-risk, lower levels of platelet (PLT) and hemoglobin (Hb), and higher expression levels of serum CysC and β₂-MG, all of which were statistically significant (P < 0.05). Logistic regression analysis showed that high expression of serum CysC, β₂-MG and adverse-risk were independent risk factors for failure of demethylation treatment in AML patients (OR >1, P < 0.05). The ROC curves showed that the AUC values of serum CysC, β₂-MG alone and combined in predicting the efficacy of demethylation therapy in AML patients were 0.788, 0.785 and 0.834, respectively.

Conclusion: The failure of demethylation therapy in AML patients is related to the high expression of serum CysC and β₂-MG, and detection of serum CysC and β₂-MG before treatment can predict the risk of demethylation therapy failure in AML patients.

The application of chimeric antigen receptor T cell (CAR-T) immunotherapy has ushered in a new era in cancer therapy, especially in the treatment of many kinds of refractory malignant tumors. The curative effect is significant for refractory/recurrent hematologic malignancies, such as acute leukemia, lymphoma, and multiple myeloma (MM). Tumor microenvironment (TME) is closely related to the efficacy and adverse reactions of CAR-T therapy. TME not only affects the activity of CAR-T cells, reduces their anti-tumor ability, but also directly involved in the occurrence and development of CAR-T cell therapy-related adverse reactions, such as cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Therefore, a deeper understanding of the role of blood TME in the process of CAR-T immunotherapy and the occurrence and development of adverse reactions is helpful for the application of CAR-T therapy in hematological malignancies. In this review, the influence of blood TME on the efficacy and adverse reactions of CAR-T immunotherapy was briefly summarized, aiming to provide evidence-based support for the clinical optimization of therapeutic regimen of refractory/recurrent hematologic malignancies.

Objective: To investigate the expression and significance of PD-1/PD-L1 in MDS blast cells, T lymphocyte cell subsets and Treg cells.

Methods: Eighty-eight MDS patients and 19 AML patients were collectd as the study subjects, and Iron deficiency anemia and healthy bone marrow donors were used as control group. The expression of PD-1/PD-L1 in MDS/AML blast cells, T lymphocyte cell subsets and Treg cells was detected by flow cytometry, and the expression level of Th1/Th2/Th17-related cytokines in peripheral serum was detected.

Results: The expression of PD-1/PD-L1 in blast cells, T lymphocyte cell subsets and Treg cells in low risk MDS group were lower than that in control group, medium and high risk MDS group and AML group（all P < 0.01）, and Th1/Th17 type cytokines were dominant. The expression of PD-1/PD-L1 in blast cells, T lymphocyte cell subsets and Treg cells of intermediate and high risk MDS group and AML group were higher than that of control group and low risk MDS group (all P < 0.01), and Th2 type and Treg type (IL-10、TGF-β) cytokines were dominant. After treatment, the differences of PD-1/PD-L1 expression were not statisticatly significant in blast cells, T lymphocyte cell subsets and Treg cells between the MDS remission group and the control group (all P >0.05). The expression of PD-1/PD-L1 in blast cells, T lymphocyte cell subsets and Treg cells in MDS non-remission group were significantly higher than that in remission group and control group (all P < 0.01).

Conclusion: The high expression of PD-1/PD-L1, dominance of Treg (IL-10、TGF-β) and Th2-related cytokines and inhibition of effector T lymphocyte cells in patients with MDS is conducive to tumor cell proliferation and immune escape, which may promote the progression of MDS disease.

Objective: To optimize the technical parameters related to the preparation of novel frozen human platelets and formulate corresponding protocol for its preparation.

Methods: Novel frozen human platelets were prepared with O-type bagged platelet-rich plasma (PRP), the key technical parameters (DMSO addition, incubation time, centrifugation conditions, etc.) of the preparation process were optimized, and the quality of the frozen platelets was evaluated by routine blood tests, apoptosis rate, platelet activation rate and surface protein expression level.

Results: In the preparation protocol of novel frozen human platelets, the operation of centrifugation to remove supernatant was adjusted to before the procedure of platelets freezing, and the effect of centrifugation on platelets was minimal when the centrifugation condition was 800×g for 8 min. In addition, platelets incubated with DMSO for 30 min before centrifugation exhibited better quality after freezing and thawing. The indexes of novel frozen human platelets prepared with this protocol remained stable after long-term cryopreservation.

Conclusion: The preparation technique of novel frozen human platelets was established and the protocol was formulated. It was also confirmed that the quality of frozen platelets could be improved by incubating platelets with DMSO for 30 min and then centrifuging them at 800×g for 8 min in the preparation of novel frozen human platelets.

Objective: To investigate the expression level of urothelial carcinoembryonic antigen 1 (lncRNA UCA1) in the bone marrow of acute myeloid leukemia (AML) patients, and to explore the clinical significance of lncRNA UCA1 expression level in AML patients.

Methods: Bone marrow samples of 50 AML patients were collected as experimental group, and bone marrow samples of 20 iron deficiency anemia (IDA) patients were collected as control group. The relevant clinicopathological characteristics of AML patients were collected. Real-time quantitative PCR (qRT-PCR) was used to detect the expression level of lncRNA UCA1 in the experimental and control groups, and the relationships between lncRNA UCA1 expression and clinical pathological characteristics and prognosis in AML patients were analyzed. Kaplan-Meier curves were used to analyze the effect of lncRNA UCA1 on the overall survival (OS) of AML patients; And Cox regression model was used to analyze the factors affecting the prognosis of AML patients.

Results: Compared with the control group, the expression level of lncRNA UCA1 was significantly elevated in patients with AML (P < 0.001); The proportion of patients with hemoglobin lower than 90 g/L in lncRNA UCA1 high expression group was significantly higher than that in lncRNA UCA1 low expression group (P =0.004); The expression level of lncRNA UCA1 was higher in M1, M2, and M4 subtypes, while it was lower in M0 and M5 subtypes, and the difference was statistically significant (P =0.009). There were no significant difference in sex, age, white blood cell (WBC) count, platelet (PLT) count, bone marrow blasts , chemotherapy regimen and efficacy, karyotype, gene mutation, and prognostic risk stratification between patients in UCA1 high expression group and those in UCA1 low expression group (all P > 0.05). The OS of patients with high expression of lncRNA UCA1 was significantly shorter than that of patients with low expression of lncRNA UCA1 (P =0.0229).

Conclusion: The expression level of lncRNA UCA1 is significantly upregulated in AML patients. High expression of lncRNA UCA1 is associated with poor clinicopathological features and poor prognosis. Therefore, lncRNA UCA1 can be used as a prognostic indicator and a potential therapeutic target for AML patients.

Objective: To investigate the expression and clinical significance of long noncoding RNA(lncRNA) HEIH in patients with acute myeloid leukemia (AML).

Methods: 50 newly diagnosed AML patients (except M3) admitted to the First Affiliated Hospital of Bengbu Medical College from January 2019 to December 2020 were included in the study, with 30 patients with non-hematological malignancies as controls. The relative expression level of lncRNA HEIH in all patients were detected, the correlation of clinical characteristics, gene mutations, FAB classification, efficacy, prognosis and overall survival (OS) of AML patients with the expression level of lncRNA HEIH were analyzed.

Results: The expression level of lncRNA HEIH in AML patients was significantly higher than that in patients with non-hematological malignancies (P <0.01). Moreover, AML patients with high white blood cell count (WBC), CEBPA and FLT3 mutations, poor efficacy, and poor prognosis often showed higher expression of lncRNA HEIH, and patients with high lncRNA HEIH expression showed a shorter overall survival (OS).

Conclusion: lncRNA HEIH shows an unique molecular biological significance in AML patients, which may provide a new approach for diagnosis, monitoring and targeted therapy of AML.