Clinical Significance of Detecting CD4T Cells in Peripheral Blood of Patients with Follicular lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the changes of CD4T lymphocytes in peripheral blood of patients with follicular lymphoma and its clinical significance.</p><p><b>METHODS</b>Blood samples were collected for detection of whole blood cells, including absolute monocyte count (AMC), absolute lymphocyte count (ALC), hemoglobin (Hb), platelet count (Plt). Age, sex, pathological grade, number of involved lymph nodes, bone marrow involvement (BMI), Ann Arbor stage, B symptoms, serum lactate dehydrogenase (LDH) and serum β-2 microglobulin (β2-MG) were recorded, the prognostic stratification was performed by using FLIPI and FLIPI-2. The T lymphocyte subsets were analyzed by flow cytometry, including the absolute number of CD4T lymphocytes (ACD4C) and the absolute number of CD8T lymphocytes (ACD8C).</p><p><b>RESULTS</b>Patients were with higher Ann Arbor stage, Hb<120 g/L, LDH greater than the upper limit of normal, the number of lymph nodes were involved> 4, the bone marrow was involvement, β2-MG levels were high FLIPI score and FLIPI-2 score, AMC level was higher (P<0.05). There were no significant differences in ACD4C levels among different groups. Patients with AMC≥0.89×10/L showed a shorter progression-free survival (PFS) and a shorter overall survival time (OS) (P=0.010,0.002) as compared with patients with AMC<0.89×10/L. The patients with ACD4C>0.16×10/L had longer progression-free survival and overall survival time, as compared with patients with ACD4C ≤0.16×10/L (P=0.016,0.012). Low ACD4C and high AMC related with shorter PFS and OS (P=0.013, 0.020). Univariate Cox regression analysis showed that age (P=0.026), bone marrow involvement (P=0.017), elevated LDH (P=0.001), β2-MG (P=0.014), FLIPI and FLIP2 score (P= 0.004 and 0.000) related with a shorter PFS. Multivariable Cox regression analysis showed that Hb (P=0.015), elevated LDH (P=0.003), β2-MG (P=0.045), bone marrow involvement (P=0.016) and FLIPI-2 score(P=0.003) related with short OS. ACD4C ≤0.16×10/L was a factor influencing prognosis of FL patients (PFS and OS) (P<0.05).</p><p><b>CONCLUSIONS</b>Low ACD4C levels relatees with poor prognosis of patients with FL, and the ACD4C levels may be an important predictor for FL disease and prognosis.</p>

Research Advances of IDH2 Gene Mutation in Acute Myeloid Leukemia / 中国实验血液学杂志

Acute myeloid leukemia (AML) is a malignant clonal hematologic disease from hematopoietic stem and progenitor cells. The isocitrate dehychogenase 2 (IDH2) gene mutation has been recently found, which may be associated with the course of AML. The incidence of IDH2 gene mutation in the patients with acute myeloid leukemia is high, especially in the AML patients with normal karyotype. Different subtypes of IDH2 mutation, or companing other molecular biology, will make different influence on clinical features and progress of patients with AML. IDH2 mutation is stable, which can be used as the test sign of AML and minimal residual disease (MRD), and for guiding the clinical treatment and predicting the progress. In this article, the research progress of IDH2 mutation in acute myeloid leukemia is reviewed.

Relationship between Calreticulin Gene Mutation and JAK2/MPL Negative Myeloproliferative Neoplasms / 中国实验血液学杂志

In 2008, WHO made the JAK2V617F gene mutation as one of the specific molecular diagnostic markers of BCR/ABL-negative myeloproliferative neoplasms (MPN). In 2013 two research teams demonstrated that whole genome sequencing technology (WGS) was used to detect calreticulin gene mutation in essential thrombocythaemia (ET) and primary myelofibrosis (PMF) patients with JAK2V617F⁻ and MPL⁻ mutations. In this review, the relationship of CALR gene mutation with MPN is briefly summarized.

Inducing-apoptosis effect of brucine on human monocytic leukemia cell line THP-1 and its mechanism / 中国实验血液学杂志

This study was aimed to investigate the inducing-apoptosis effect of brucine on human monocytic leukemia cell line THP-1 cells and its possible mechanism. The inhibition effect of brucine on growth of THP-1 cells was measured by CCK-8 method. Morphological changes of THP-1 cells treated with brucine was detected by acridine orange/ethidium bromide (AO/EB)double staining. Annexin-V/PI double labeling method was used to assay the apoptosis rate of THP-1 cells. The effect of brucine on THP-1 cell cycle distribution was detected by PI single staining. RT-PCR was used to detect the expression of BCL-2 and BAX. The results showed that the brucine could inhibit the THP-1 cell growth in concentration and time-dependent manners at the range of 50 to 400 µg/ml. The cells stained with AO/EB revealed that the brucine induced the nuclear chromatin condensation. After the THP-1 cells were treated with brucine of 400µg/ml for 48 hours, most nucleic were stained as orange-red, and condensed, displaying the late apoptotic cell morphology. Annexin-V/PI detection showed that brucine could induce apoptosis of THP-1 cells in a concentration-dependent manner. Compared with the control group, more cells in brucine-treated group were arrested at G0/G1 phase in a concentration-dependent manner. RT-PCR detection revealed that the expression of BCL-2 was down-regulated strikingly and BAX was up-regulated. It is concluded that brucine can efficiently inhibit cell growth and block THP-1 cells in G0/G1 phase. The mechanism of THP-1 cell apoptosis induced by brucine may be related to the inhibition of BCL-2 and activation of BAX.

Th17 cells and aplastic anemia / 中国实验血液学杂志

During the past few years, a novel family of CD4⁺T cell lineage was detected and named as Th17 cells because of its unique ability expressing IL-17, which also can produce IL-17A, IL-17F, IL-21, IL-22 and IL-26. Some cytokines, such as TGF-β, IL-6, L-23 may promote the differentiation of Th17 subset, whereas some cytokines, such as IL-21, IL-2, IFN-γ, may have inhibitory effects. Th17 cells serving as immune effectors play an important role in autoimmune diseases caused by chronic inflammation injury. More and more studies confirmed that Th17 cells have closely correlations with the development of aplastic anemia, and may be a new target in the diagnosis, therapy, prognosis and prophylaxis of aplastic anemia.

Effect of brucine on secretion function and proliferation capability of T lymphocytes in patients with aplastic anemia / 中国实验血液学杂志

The aim of this study was to investigate the effect of brucine on secretion of TNF-α, IFN-γ, IL-4 and proliferation of T lymphocytes in patients with aplastic anemia (AA), and to explore its mechanism. Peripheral blood T lymphocytes from 10 patients with AA and 10 healthy volunteers were isolated, purified and cultured. T lymphocytes from the patients were divided into 0, 100, 200 and 400 µg/ml brucine-treated groups. T lymphocytes from healthy volunteers were used as control group. After being cultured for 72 hours, the levels of TNF-α, IFN-γ, IL-4 in the supernatant of cultured T lymphocytes from AA patients were detected by ELISA, and the proliferation of T lymphocytes from AA patients was detected by MTT. The results showed that compared with the normal control group, the levels of TNF-α and IFN-γ in the culture supernatant significantly increased, and IL-4 was significantly decreased. The levels of TNF-α and IFN-γ in the culture supernatant of brucine treated groups were lower, and were dependent on the concentration of brucine. However, the levels of IL-4 were found to be not obviously changed. The inhibition rate of T lymphocytes in 100, 200 and 400µg/ml brucine-treated groups were (13.61 ± 4.31)%, (14.28 ± 4.31)% and (15.12 ± 4.56)% respectively, among which the differences were not statistically significant. It is concluded that the brucine can reduce the levels of TNF-α and IFN-γ through inhibiting the proliferation of T lymphocytes in AA patients, which provides experimental basis for therapy of AA patients.

Apoptosis-inducing effects of brucine on human chronic myeloid leukemia cell line K562 / 中国实验血液学杂志

To investigate the apoptosis-induction effect of brucine on human chronic myeloid leukemia cell line K562 cells, K562 cells were exposed to various dosages of brucine. MTT method was used to assayed the growth inhibition effect of brucine on K562 cells. The apoptosis of K562 cells was detected by acridine orange/ethidium bromide (AO/EB) double staining, Annexin-V/PI double labeling method and DNA agarose gel electrophoresis. The results showed that brucine could remarkably inhibit the K562 cell growth in a concentration-dependent and time-dependent manners at the range of 50 to 400 µg/ml, and its most significant inhibition was observed at 400 µg/ml for 72 hours and the inhibition rate was 94.0%. Staining of cells with AO-EB revealed that brucine induced nuclear chromatin condensation. After the K562 cells were treated with the brucine of 400 µg/ml for 72 hours, the most of the nucleus were orange stained and condensation-like or bead-like showing apoptotic morphology. The K562 cells treated with brucine of different concentrations (50, 100, 200, 400, 800 µg/ml) for 72 hours, Annexin-V/PI detection showed brucine could induce apoptosis of K562 cells, and apoptosis rate increased gradually with increasing concentration of drugs. The K562 cells treated with brucine of 400 µg/ml for 72 hours displayed typical ladder strap in DNA gel electrophoresis. It is concluded that brucine can efficiently inhibit cell growth and induce apoptosis of K562 cells with dose-dependent manner in concentrations of 50 - 400 µg/ml.