Research Progress on Artemisinin and Its Derivatives against Hematological Malignancies / 中国结合医学杂志

Although current therapeutic methods against hematological malignancies are effective in the early stage, they usually lose their effectiveness because of the development of drug resistances. Seeking new drugs with significant therapeutic effects is one of the current research hotspots. Artemisinin, an extract from the plant Artemisia annua Linne, and its derivatives have excellent antimalarial effects in clinical applications as well as excellent safety. Recent studies have documented that artemisinin and its derivatives (ARTs) also have significant effects against multiple types of tumours, including hematological malignancies. This review focuses on the latest research achievements of ARTs in the treatment of hematological malignancies as well as its mechanisms and future applications. The mechanisms of ARTs against different types of hematological malignancies mainly include cell cycle arrest, induction autophagy and apoptosis, inhibition of angiogenesis, production of reactive oxygen species, and induction of differentiation. Additionally, the review also summarizes the anticancer effects of ARTs in many drug-resistant hematological malignancies and its synergistic effects with other drugs.

Expression of TFPI-2 gene and its promoter methylation in acute myeloid leukemia / 中国实验血液学杂志

The aim of this study was to detect the mRNA expression of tissue factor pathway inhibitor-2 ( TFPI-2) and its methylation in bone marrow mononuclear cells from acute myeloid leukemia (AML) patients and to explore its significance in AML. Bone marrow mononuclear cells were isolated from newly diagnosed AML patients (n = 33), complete remission AML patients (n = 19), relapsed/refractory AML patients (n = 12) and iron deficiency anemia patients (control group, n = 15). Expression of TFPI-2 mRNA was detected with real-time quantitative PCR (RT-PCR) and the methylation of CpG island in its promoter was detected with methylation-specific PCR (MSP). The results showed that the expression of TFPI-2 mRNA in newly diagnosed AML, complete remission AML and relapsed/refractory AML patients was much lower than that in the controls (P < 0.05). Furthermore, its expression in relapsed/refractory AML patients was lower than that in newly diagnosed AML patients (P = 0.006). Compared with complete remission AML patients, the expression of TFPI-2 mRNA in newly diagnosed AML patients was significantly reduced (P = 0.030). The percentage of TFPI-2 promoter methylation in AML patients was 64.63% (42/64). In newly diagnosed AML group, complete remission AML group and relapsed/refractory AML group,the percentages of TFPI-2 promoter methylation were 66.67% (22/33), 52.63% (10/19) and 83.33% (10/12) (P > 0.05), respectively. The optical density ratio of TFPI-2 mRNA expression was 0.165 (0.005-2.099) in methylated AML patients, and 0.597 (0.011-2.787) in unmethylated AML patients (P < 0.05). Methylation of TFPI-2 gene promoter was not detected in control patients. After 2 courses of chemotherapy, the level of TFPI-2 mRNA was much higher in the CR group than that in the non-CR group (P < 0.05). It is concluded that the down-regulation or silence of TFPI-2 gene potentially results from its promoter methylation, and the expression level of TFPI-2 and the methylation status of its promoter may be used as indicators of risk stratification and evaluation of disease progress.

Expressive changes of CD4(+)T cell subset transcription factors in patients with aplastic anemia, myelodysplastic syndrome and acute myeloid leukemia and their clinical significances / 中国实验血液学杂志

This study was aimed to compare the expressions of specific transcription factors of CD4(+) T cell subset ( T-bet, GATA-3, RORγt and FoxP3 mRNA) in peripheral blood of patients with aplastic anemia(AA), myelodysplastic syndrome(MDS), and acute myeloid leukemia(AML), and investigate their immune status and pathogenesis, so as to provide experimental basis for the choice of clinical treatment. The expression of T-box (T-bet), GATA-3, ROR-γt and Foxp3 mRNA in PBMNC were examined by RT-PCR in 42 cases of MDS, including 22 refractory anemia(MDS-RA) and 20 refractory anemia with excess blasts (MDS-RAEB), in 23 cases of AA, 17 cases of AML patients and 16 healthy volunteers respectively. The results indicated that, compared with normal control group, expressions of T-bet and RORγt mRNA in AA patient group were significantly higher (P < 0.01), expression levels of GATA3 Foxp3 mRNA were lower (both P < 0.01). There was no significant difference in expression of T-bet and GATA3 mRNA between MDS group and normal control group, but the expression levels of Foxp3 and RORγt mRNA were higher than those in normal controls (P < 0.05); T-bet and RORγt in MDS-RA group were higher than those in the normal controls(P < 0.01), and GATA3 expression significantly reduced (P < 0.05), however, there was no significant difference in expression of Foxp3 between MDS-RA and the controls. Expression levels of T-bet and RORγt mRNA in patients with MDS-RAEB and AML were lower than those in normal controls (P < 0.05), but the expression levels of GATA3 and Foxp3 mRNA were significantly higher than those in normal controls (P < 0.01). It is concluded that the transcription factor expressions are different in PBMNC of patients among these three diseases. Immune-mediated excessive apoptosis may play an important role in pathogenesis, bone marrow failure in patients with AA and MDS-RA, and abnormal clones of immature cells may be one of main reasons for bone marrow failure in AML and late stage of MDS.

Clinical significance of mast cells and IL-9 in B-NHL / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of mast cells and interleukin-9 (IL-9) in B-cell non-Hodgkin lymphoma (B-NHL) development and its clinical significance.</p><p><b>METHODS</b>The expression level of CD117 in tumor tissues of 32 B-NHL patients was determined by Western blot. The infiltration of CD117⁺ mast cells (MCs) in human B-NHL tumor tissues was observed by immunohistochemistry staining. To evaluate the correlations between the data from CD117⁺ MCs and biological markers of human B-NHL, a Spearman correlation coefficient (rs) was calculated. IL-9 levels in sera of B-NHL patients were measured by ELISA. Effects of IL-9 on expressions of functional genes of mouse bone marrow-derived mast cells (BMMCs) were detected by real-time quantitative RT-PCR.</p><p><b>RESULTS</b>The expression of CD117 was upregulated significantly in human B-cell NHL involved tissues when compared with that of controls (0.0551±0.0064 vs 0.0192±0.0072, P<0.01). Infiltration of more CD117⁺ MCs was found in tissues from B-cell NHL subjects compared with that of controls. IL-9 level in serum samples from patients with B-cell NHL was higher than that from healthy controls. Addition of rIL-9 to the culture gave rise to increase in the purity of mouse BMMCs in the first three weeks. In vitro culture experiments showed that the addition of IL-9 could induce the differentiation of mouse BMMC and the expressions of MC-related genes, including CD117, Fcer1α, Mcpt1 and Mcpt5.</p><p><b>CONCLUSION</b>Our study showed that IL-9 promoted immune response mediated by MCs, and probably played important roles in B-NHL growth. Pharmacological or targeted inhibition of mast cells or IL-9 activity may provide new strategy for B-cell NHL therapy.</p>

Changes and clinical significance of CD8(+) T cell subset in patients with aplastic anemia, myelodysplastic syndrome and acute myeloid leukemia / 中国实验血液学杂志

This study was purposed to detect the balance and the activity change of cytotoxic T cell subsets in aplastic anemia (AA) patients, myelodysplastic syndrome (MDS) patients and acute myeloid leukemia (AML) patients, and to explore the cellular immune mechanism for abnormal hematopoiesis of the three diseases, so as to provide experimental basis for the choice of clinical treatment. The proportion of the cytotoxic T cells and part of the T-cells subsets in peripheral blood were detected by flow cytometry in 35 cases of MDS, including 19 refractory anemia (MDS-RA), 16 refractory anemia with excess blasts (MDS-RAEB), 17 AA, 15 AML patients and 10 normal donors respectively. The results showed that compared with the control group, the percentage of Tc1, Tc1/Tc2, CD8(+)HLA-DR(+), CD3(+)CD8(+)CD28(+), CD8(+)CD45RO(+) cells was significantly higher and the percentage of CD8(+)CD45RA(+) was significantly lower in AA and MDS-RA group. There was no difference in the percentage of Tc2 cells between AA/MDS-RA and normal controls; the percentage of CD8(+)CD45RO(+) cells was significantly higher and the percentage of Tc1, CD3(+)CD8(+)CD28(+), CD8(+)HLA-DR(+) was significantly lower in MDS-RAEB group, the percentage of CD8(+)CD45RA(+) was lower but the difference was not significant, and there was no difference in the percentage of Tc, Tc1/Tc2 cells between MDS-RAEB group and the control group. The percentage of Tc2 cells was significantly higher and the percentage of other parameters was significantly lower in AML group than those of normal controls. It is concluded that the cellular immune statuses in AA, the different stages of MDS and AML are different. In AA and the early stage of MDS, the balance of Tc1/Tc2 shifts to Tc1, and the activation of T-cell subsets increases. In the late stage of MDS and AML, the balance of Tc1/Tc2 shifts to Tc2, the activation of T-cell subsets decreases. The former may be closely related to bone marrow failure while the latter may be one of the important mechanisms in which the malignant clones escape from immune effect.

The study of IL-18 and IL-18BP balance in aplastic anemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the role of IL-18 and IL-18BP balance in aplastic anemia (AA).</p><p><b>METHODS</b>A total of 29 AA patients and 22 controls were recruited in present research. The expressions of IL-18 and IL-18BP were measured by enzyme-linked immunosorbent assay (ELISA). The mRNA expressions of IL-18 and IL-18BP were measured in all subjects using real-time quantitative polymerase chain reaction (RT-PCR).</p><p><b>RESULTS</b>The levels of the IL-18 in plasma of AA and normals were (365.5 ± 160.6) pg/ml and (175.9 ± 92.8) pg/ml (P < 0.01); and the expression of IL-18 in severe AA patients (441.3 ± 116.9) pg/ml were higher than that in non-severe AA patients (326.4 ± 167.0) pg/ml (P < 0.05). The level of IL-18BP was increased in plasma of AA (1788.6 ± 523.8) pg/ml than in normals (1083.6 ± 489.6) pg/ml (P < 0.05). But the ratio of IL-18/IL-18BP in AA patients was much higher than that in controls (P < 0.05). RT-PCR revealed the levels of IL-18 and IL-18BP mRNA were up-regulated in AA patients when compared to controls, but the ratio of IL-18/IL-18BP was significantly elevated in AA patients.</p><p><b>CONCLUSION</b>IL-18/IL-18BP imbalance may play an important role in pathogenesis of AA and regulating the balance of IL-18 and IL-18BP may be a therapeutic approach to AA.</p>

Secretion of IL-18 and IL-18 binding protein from splenocytes of ITP patients in vitro / 中国实验血液学杂志

This study was aimed to investigate the expression and clinical significance of IL-18, IL-18 binding protein (IL-18BP), IFN-γ and IL-4 secreted from splenocytes of patients with idiopathic thrombocytopenic purpura (ITP) in vitro. Spleen mononuclear cells (MNC) were prepared by using routine sterile method, and were cultured in RPMI 1640 complete medium containing 10 µg/ml PHA, 10% fetal calf serum at 37°C and 5% CO2. The levels of IFN-γ, IL-4, IL-18 and IL-18BP secreted from MNC of ITP patients and normal controls were determined after culture for 48 hours. The results showed that after culture of spleen MNC for 48 hours, the levels of IL-18 and IFN-γ were significantly higher in patients with ITP than that in controls, but the levels of IL-18BP was not significantly elevated in ITP patients. The level of IL-4 was below the detectable limit of the assay used. It is concluded that imbalance between IL-18 and IL-18BP may play an important role in pathogenesis of ITP, and regulation of balance between IL-18 and IL-18BP may be a therapeutic approach against ITP.