A new 9,19-cycloartane glycoside from Asplenium ruprechtii / 中国中药杂志

Chemical constituents of water extracts of Asplenium ruprechtii were investigated. Five compounds were isolated by silica gel, Sephadex LH-20 gel column chromatographies and preparative HPLC, and their structures were identified by various spectral analyses as aspleniumside G(1), trans-p-coumaric acid(2), trans-p-coumaric acid 4-O-β-D-glucoside(3), cis-p-coumaric acid 4-O-β-D-glucoside(4), and(E)-ferulic acid-4-O-β-D-glucoside(5). Among them, compound 1 is a new 9,19-cycloartane glycoside.

High expression level of TAL1 gene in newly diagnosed acute myeloid leu-kemia / 中国病理生理杂志

AIM:To investigate the characteristic of T-cell acute lymphocytic leukemia 1 (TAL1) gene expres-sion in acute myeloid leukemia (AML) cell lines and in primary AML cells from de novo AML patients with different sub-types. METHODS:Real-time PCR was used to determine the expression of TAL1 mRNA in acute leukemia cell lines (Jurkat, CCRF-CEM, HL-60 and NB4 cell lines) and peripheral blood mononuclear cells from 47 newly diagnosed AML patients. Twelve healthy individuals were served as healthy control group. RESULTS:A significantly increased level in TAL1 mRNA was found in AML cell lines (HL-60 and NB4), T-cell acute lymphacytic leukemia (T-ALL) cell lines (Jur-kat, CCRF-CEM) and primary AML cells compared with the healthy controls. Over-expression of TAL1 was found in all detected AML subtypes, the highest level of TAL-1 mRNA was found in AML-M1 and AML-M5 subtype ( P <0.05). CONCLUSION:High expression of TAL1 in AML might influence the differentiation and proliferation of myeloid cells, further investigation needs to confirm whether it would be as a biomarker for pathogenesis of AML.

Screening and evaluation of Smo-siRNA targeted to inhibition of Molt-4 cell proliferation / 中国组织工程研究

BACKGROUND: Studies have shown that the occurrence and development of T lymphocytic leukemia is related to the abnormality of Hedgehog pathway. The Smo gene is a key gene in this signaling pathway and controls the transmission of Hedgehog signaling into the cell membrane. OBJECTIVE: To design and screen a highly efficient and specific Smo-siRNA which is able to downregulate the Smo gene expression in Molt-4 cells, thereby inhibiting the Molt-4 cells proliferation and inducing apoptosis. METHODS: (1) Smo-siRNAs numbered 1, 2 or 3, and the scrambled non-siRNA control (SC) were obtained by chemosynthesis. Untreated and sc-treated cells were used as controls. (2) Smo expression levels in Molt-4 cells were analyzed using qRT-PCR at 24, 48, 72 hours after siRNAs delivered by NuclefectorTM.Cell proliferation in vitro was assayed by the cell counting kit-8.The morphology and percentage of apoptotic cells were revealed by Hoechst33258 staining and flow cytometry, respectively. RESULTS AND CONCLUSION: (1) Smo-siRNAs were successfully transferred into Molt-4 cells, and exhibited best silencing results. After transfection with Smo-siRNA1, the mRNA level of Smo was significantly reduced (P < 0.05), and the lowest level was at 48 hours after transfection. (2) Cell proliferation of Molt-4 cells was significantly inhibited by Smo-siRNA at 24 hours after transfection. (3) Hoechst staining results showed morphological changes of Molt-4 were in accordance with those of apoptotic cells. (4) The apoptotic rate was significantly increased in the Smo-siRNA group compared with the control group (P < 0.05). Findings from this study showed that suppression of Smo by RNA interference could effectively inhibit proliferation and induce apoptosis in Molt-4 cells, indicating that Smo-siRNA as gene targeted therapy or synergistic treatment has therapeutic potential in T-cell malignancies.

Influence of diethyl sulfate (DES) mutagenesis on growth properties and pigment secondary metabolites of Phellinus igniarius / 中国中药杂志

The diethyl sulfate (DES) mutagenesis was chosen for the mutagenic treatment to Phellinus igniarius, and the relationship of mutagenesis time and death rate was investigated with 0.5% DES. The differences of mycelial growth speed, liquid fermentation mycelia biomass, morphology and pigment classes of secondary metabolites production speed and antioxidant activities of metabolite products were discussed. The study displayed that DES mutagenesis could change mycelial morphology without obvious effect on mycelium growth, and the DES mutagenesis improved antioxidant activities of the active ingredients of P. igniarius and had more antioxidant activity of hypoxia/sugar PC12 nerve cells than that of P. igniarius.

Separation and identification of endophytic fungi from desert plant Cynanchum komarovii / 中国中药杂志

<p><b>OBJECTIVE</b>The research aimed to investigate the entophytic fungal community of Cynanchum Komarrovii, including the biodiversity in different organs and the correlations with ecological environment. Endophytic fungi with patent bioactivity were also rapidly screened.</p><p><b>METHOD</b>PDA medium was used to isolate and purify the endophytic fungi from C. komarovii living in Shaanxi and Ningxia district, respectively. The strains were identified based on the morphological characteristics of the fungi and similarity of 5.8S gene and internal transcribed spacer (ITS) sequence. Pyriculaia oryzae model was applied to preliminarily screen the active fungi.</p><p><b>RESULT</b>Ninety-four strains of endophytic fungi were isolated and identified to 9 species, 13 genera, 9 families and 6 orders, among them, 47 strains were from the plants living in Ningxia. And then, 5 of them were isolated from roots, 14 from branches, and 28 from leaves. They were identified belonging to 8 species, 9 genera, 5 families and 4 orders. Additionally, 47 strains were from the plants living in Shaanxi. 16 were isolated from the roots, 18 from branches, 13 from leaves. They were identified belonging to 5 species, 8 genera, 6 families and 4 orders. By preliminary screening, 18 strains of endophytes completely inhibited the germination of conidium, which showed a potential bioactivity for these fungi. Both N4 and S17 strains had stronger growth inhibition effect.</p><p><b>CONCLUSION</b>Endophytic fungi from desert plant C. komarovii have the feature of diversity. Different geographical environment and type of organizations lead to the significant difference on the quantity and the species composition. Most of fungi in Ningxia C. komarovii distribute in leaves. However, most of those in Shaanxi C. komarovii distribute in stems and leaves. It also indicated that endophytes from C. komarovii had a strong antifungal activity.</p>

Determination of chrysosplenetin, metabolic inhibitor of artemisinin, in rat plasma by UPLC-ms/MS and study on its pharmacokinetics / 中国中药杂志

<p><b>OBJECTIVE</b>The study aimed to develop the assay of chrysosplenetin (CHR), a metabolic inhibitor of artemisinin by UPLC-MS/MS in rat plasma and investigate the pharmacokinetics parameters of CHR.</p><p><b>METHOD</b>The plasma samples were precipitated by acetonitrile to remove the proteins. Separation was carried out on a Shim-pack XR-ODS C,18(2. 0 mm x 100 mm, 2. 2 micromp.m) column using a mobile phase containing methanol-0. 1% formic acid (87:13) using by diazepam as internal standard. Mass spectrometer with electrospray ionization (ESI) operated in the positive ion mode was used for analysis. Total analysis time was 2 min.</p><p><b>RESULT</b>The assay was linear in the range 5-5 000 microg L-1 (r =0. 999 3) with recoveries in the range from 69. 0% to 81.2% and satisfied inter-, intra- precision and accuracy. CHR after oral administration is not easy to absorb with double or multimodal peak phenomenon. The t1/2 of CHR after intravenous injection was very short and that of low, medium, and high dosage was (17. 01 +/- 8. 06) , (24. 62 +/- 4. 59), (28. 46+/- 4. 63) min, respectively.</p><p><b>CONCLUSION</b>The developed method was special, rapid, and sensitive for determination of CHR pharmacokinetics. [Key words] UPLC-MS/MS; chrysosplenetin; pharmacokinetics; plasma; rat</p>

Effect of granulocyte colony stimulating factor on myeloid-derived suppressor cells in the bone marrow and peripheral blood: a preliminary study / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of granulocyte colony stimulating factor (G-CSF) on myeloid-derived suppressor cells (MDSCs) in the bone marrow and peripheral blood, and explore the relationship between MDSC and graft-versus-host disease (GVHD).</p><p><b>METHODS</b>Bone marrow, peripheral blood and peripheral blood stem cells were obtained from 12 healthy hemopoietic stem cell donors before and on day 5 after G-CSF mobilization. Flow cytometry was employed to examine the number of MDSC, and the relationship between MDSC number and the incidence of GVHD was analyzed.</p><p><b>RESULTS</b>In normal physiological conditions, MDSC could be detected in the peripheral blood and bone marrow with a cell percentages of (1.35±0.35)% and (2.44±1.11)%, respectively, showing a significantly higher cell percentage in the bone marrow (P=0.015). On the 5th day after G-CSF mobilization, the percentage of MDSCs increased to (4.01±1.82)% in the peripheral blood and to (4.38±2.19)% in the bone marrow, showing no significant difference between them (P=0.083). The mobilization caused a significant increase in the number of MDSCs in the peripheral blood (P=0.047) but not in the bone marrow (P=0.761). The number of MDSCs in the collected samples showed a significant inverse correlation to the incidence of GVHD (P=0.048).</p><p><b>CONCLUSIONS</b>MDSCs are present in the peripheral blood and bone marrow of healthy donors, with a greater number in the bone marrow. G-CSF can mobilize the MDSCs from the bone marrow to the peripheral blood to increase number of MDSCs in the peripheral blood, which may contribute to a lowered incidence of GVHD in hematopoietic stem cell transplantation (HSCT).</p>

Clinical implications of HLA-G protein expression in acute leukemia / 南方医科大学学报

<p><b>OBJECTIVE</b>To detect the expression of membrane-bound HLA-G (mHLA-G) and serum HLA-G (sHLA-G) in acute leukemia patients and investigate the correlation between HLA-G expression and the occurrence and development of acute leukemia.</p><p><b>METHODS</b>Enzyme-linked immunosorbent assay and flow cytometry were used to detect the expression levels of sHLA-G and mHLA-G in 40 newly diagnosed leukemia cases, 10 refractory and relapsed leukemia cases, and 30 leukemia cases receiving chemotherapy. Ten normal individuals served as the normal control group.</p><p><b>RESULTS</b>The mean serum level of sHLA-G in normal individuals was 5.87±2.07 ng/ml, as compared to 10.05±6.58 ng/ml in newly diagnosed leukemia patients and 12.32±5.85 ng/ml in refractory and relapsed cases. The mean level of mHLA-G in normal individuals, newly diagnosed cases, and refractory and relapsed cases were (0.29±0.20)%, (0.60±0.44)%, and (0.77±0.41)%, respectively. The mean levels of sHLA-G and mHLA-G were significantly higher in the newly diagnosed cases than that in the normal controls (P<0.05), and significantly higher in patients before chemotherapy than in those with complete remission after chemotherapy (P<0.05).</p><p><b>CONCLUSION</b>HLA-G expression levels might influence the treatment outcomes and can serve as a prognostic factor for acute leukemia.</p>

Quantitative real-time PCR for detecting the expression levels of TCR Vgamma subfamilies in patients with graft-versus-host disease following allogenic hematopoietic stem cell transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of T cell receptor (TCR) Vgamma genes in patients with graft-versus-host disease (GVHD) after allogenic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>The expression levels of the TCR Vgamma I-III genes in peripheral blood mononuclear cells (PBMNCs) from 18 patients with GVHD following allo-HSCT were determined using real-time fluorescence quantitative PCR, with 12 healthy individuals serving as the normal controls.</p><p><b>RESULTS</b>The expression level of TCR Vgamma II gene in the PBMNCs from patients with GVHD was significant lower than that in the normal controls. The expression patterns of TCR Vgamma I-III subfamilies also underwent alterations in patients with GVHD, and the expression level of TCR Vgamma II gene was significantly lower than that of TCR Vgamma I gene or TCR Vgamma III gene.</p><p><b>CONCLUSION</b>The low expression of TCR Vgamma II subfamily might be related to the pathogenesis of GVHD in patients receiving allo-HSCT.</p>

Study on relationship between chemical castration and thymic function following hematopoietic stem cell transplantation / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the impact of luteinizing hormone-releasing hormone (LHRH) on the protection of thymic function after allogenic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>Murine model of MHC mismatched allogeneic HSCT (C57BL/6-->BALB/c) was established. The severity of acute graft-versus-host-disease (GVHD) was assessed according to a clinical scoring system. The intra-cellular levels of IFN gamma, TNFalpha and IL-1 beta in thymocyte were analyzed by protein array and thymic function by quantification of signal-joint TCR rearrangement excision circles (sjTRECs).</p><p><b>RESULTS</b>All recipients in group A (allogeneic mice), B (allogeneic LHRH castrated-mice) and C (syngenic mice) achieved hematopoietic reconstitution. White blood cell (WBC) over 1.0 x 10(9)/L in groups A, B and C were on day (11.2 +/- 1.4), day (9.8 +/- 0.6) and day (9.7 +/- 0.7), respectively (P = 0.003, 0.002). The onset of acute GVHD in group B was (14.1 +/- 0.7) d and in group A was (11.4 +/- 1.2) d (P = 0.000). All mice in groups A and B developed acute GVHD. No mice occurred aGVHD in group C. The average scores of acute GVHD in groups A and B were (9.1 +/- 0.7) and (5.1 +/- 1.0), respectively (P = 0.000). The levels of IFN gamma, TNFalpha and IL-1 beta in control group were (2.3 +/- 2.5) ng/ml, (1.7 +/- 1.1) pg/ml and (1.8 +/- 1.2) pg/ml, respectively. The IFN gamma levels in groups A, B and C were (10.5 +/- 2.1) ng/ml, (6.7 +/- 2.1) ng/ml and (5.2 +/- 3.3) ng/ml, TNFalpha levels were (7.0 +/- 2.6) pg/ml, (4.3 +/- 0.8) pg/ml and (3.0 +/- 1.8) pg/ml, and IL-1 beta levels were (24.9 +/- 9.0) pg/ml, (17.4 +/- 3.9) pg/ml and (10.8 +/- 3.1) pg/ml, respectively. There were significant differences in the levels of cytokines between group A and the control group (P = 0.000, 0.000, 0.000). The levels of cytokines in group B were significantly higher than those in control group (P = 0.000, 0.003, 0.000). The levels of IFN gamma and IL-beta in group C were significantly higher than those of in control group (P = 0.015, 0.013), and so did in group A than in group B (P = 0.002, 0.002, 0.004), and in group A than in group C (P = 0.000, 0.000, 0.000). The analysis of linear regression showed that the average levels of IFN gamma and TNFalpha paralleled with aGVHD scores (r(2) = 0.359, P = 0.045; r(2) = 0.228, P = 0.019). The average sjTRECs copies/1000 PBMNCs were (39.4 +/- 44.7) in the control group and (12.3 +/- 13.0), (58.0 +/- 71.8) and (19.6 +/- 14.6) in groups A, B and C, respectively. There was no significant difference in the multiple comparisons of peripheral blood levels of sjTRECs among these four groups (P = 0.468).</p><p><b>CONCLUSION</b>IFN gamma, TNFalpha and IL-1 beta might be involved in the damage to the thymus by acute GVHD. Sex steroid inhibitor can not only reduce the severity of thymic damage after allo-HSCT, but also reduce the severity of aGVHD and the mechanism might be associated with the reduction of intra-cellular levels of IFN gamma in thymocyte.</p>

Research Progress on Trehalose Synthase / 微生物学通报

Trehalose is a naturally occurring non-reducing disaccharide. This sugar has been known as a protector of biomembrane and macromolecules. And it has been widely used in foods, medicines, cosmetics. Trehalose synthase (TreS) is an intramolecular transglycosylase. It can catalyze the conversion of maltose into trehalose in one step. As the simple protocol and cheap substrate, it is very prospect in the industrial production. Here we have discussed the properties, catalytic mechanism and gene-engineering of TreS, and also proposed some solutions to its present research problems.

Role of membrane-bound human leukocyte antigen G in inducing immune tolerance after allogeneic hematopoietic stem cell transplantation / 南方医科大学学报

<p><b>OBJECTIVE</b>To evaluate the role of human leukocyte antigen G (HLA-G) in the better effect of allogenetic bone marrow transplantation than that of peripheral blood stem cell transplantation.</p><p><b>METHODS</b>Flow cytometry was used to detect the expression of membrane-bound HLA-G (mHLA-G) on donor peripheral blood (PBC) or bone marrow (BM) mononuclear cells. The levels of soluble HLA-G (sHLA-G) in the plasma and bone marrow fluid were determined using enzyme-linked immunosorbent assay (ELISA) before and after granulocyte colony-stimulating factor (G-CSF) mobilization.</p><p><b>RESULTS</b>The mean levels of mHLA-G after G-CSF mobilization in the PBC and BM were significantly higher than that before G-CSF mobilization (P=0.001 and 0.000), but the plasma levels of sHLA-G showed no significant changes after the mobilization (P=0.279). The mean levels of sHLA-G in the BM fluid significantly increased (P=0.002) to a level higher than that in the PBC after G-CSF mobilization (P=0.004).</p><p><b>CONCLUSION</b>HLA-G plays an important role in immune tolerance after hematopoietic stem cell transplantation with G-CSF mobilization.</p>

Ovarian steroid cell tumor, not otherwise specified: a clinicopathologic study / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the clinicopathologic features, diagnostic criteria, differential diagnosis and treatment options of ovarian steroid cell tumor, not otherwise specified (NOS).</p><p><b>METHODS</b>Light microscopy and immunohistochemical study was carried out in 8 cases of ovarian steroid cell tumor, NOS. The literature was reviewed.</p><p><b>RESULTS</b>The 7 cases of benign ovarian steroid cell tumor, NOS were composed mainly of polygonal cells with granular eosinophilic cytoplasm and larger cells with vacuolated cytoplasm. They resembled the architecture of normal adrenal gland, with formation of cell nests and trabeculae. The single case of malignant ovarian steroid cell tumor had evidence of significant cellular pleomorphism, haemorrhage and coagulative tumor necrosis. The mitotic count measured about 7 per 10 high-power fields. Immunohistochemical study showed that the tumor cells expressed calretinin and alpha-inhibin. Differential diagnosis included oxyphilic granulosa cell tumor, thecoma, Sertoli cell tumor and clear cell carcinoma. The treatment options of benign ovarian steroid cell tumor, NOS was local excision or ipsilateral salpingo-oophorectomy, while the malignant counterpart should be treated with a combination of surgery and chemotherapy, including administration of GnRH agonist.</p><p><b>CONCLUSIONS</b>Ovarian steroid cell tumor, NOS, is the most common type of ovarian steroid cell tumors. Most of which are associated with a benign clinical outcome. Immunohistochemistry is an important adjunct for diagnosis. The treatment options of ovarian steroid cell tumor, NOS depend on its malignant potential.</p>

Construction of rat bdnf gene lentiviral vector and its expression in mesenchymal stem cells / 生物工程学报

Recently, mesenchymal stem cells (MSCs) have been one of the target cells of gene engineering. To construct the lentiviral (LV) vectors carrying the brain-derived neurotrophic factor (Bdnf) gene, the rat mesenchymal stem cells (rMSCs) were infected and finally the Bdnf gene-modified rMSCs was obtained. The CDS region of the rat Bdnf gene was obtained with reverse transcriptase-polymerase chain reaction (RT-PCR), and the transfer plasmid (PNL-BDNF-IRES2-EGFP) of the LV vector was constructed. The three plasmids of LV vector: PNL-BDNF-IRES2-EGFP, HELPER, and VSVG were cotransfected to 293T cells to produce the LV vectors, which enabled the coexpression of the Bdnf gene and the enhanced green fluorescent protein (Egfp) gene. rMSCs were separated from the bone marrow of 2-month-old F344 rats, cultured in vitro, and identified. rMSCs were infected by the LV vectors that were produced already and were identified with fluorescent microscope, RT-PCR, immunocytochemical staining, and western blot. The result of sequencing showed that the sequence of the cloned Bdnf gene was consistent with that reported in the GenBank. The PNL-BDNF-IRES2-EGFP plasmid that was identified showed the correct sequence. After the 3 plasmids of LV vectors were cotransfected to the 293T cells, considerable green fluorescence in 293T cells was observed under the fluorescent microscope; the supernatant was collected and concentrated using ultracentrifugation, and the titer of the replication-defective LV vector particles measured was found to be 6.7 x 10(7) TU/mL. After the constructed LV vectors infected the rMSCs, the results obtained using RT-PCR, immunocytochemical staining, and western blot showed that the expression of BDNF in the Bdnf-rMSCs group (experimental group, EG) was significantly higher than that in the PNL-IRES2-EGFP-rMSCs group (mock group, MG) and the rMSCs group (control group, CG) at both mRNA and protein levels. LV vectors carrying the Bdnf gene were constructed successfully. The Bdnf gene-modified rMSCs could express BDNF to a higher degree. This greatly facilitates the next step in the study, such as the long period of therapeutic observation of cerebral ischemia with Bdnf gene-modified rMSCs.

Thymic recent output function in patients with B-cell lymphocytic malignancies / 中国实验血液学杂志

The aim of the study was to analyze the naive T cell level of thymic recent output in patients with B-cell malignancies, thereby to evaluate the potential T-cell function. Quantitative analysis of T-cell receptor rearrangement excision circles (TRECs) in DNA of peripheral blood mononuclear cells from 61 cases of B-cell lymphocytic malignancy (including 20 cases of adult B-ALL, 6 case of childhood B-ALL, 4 cases of B-CLL, 17 cases of B-NHL and 14 cases of MM) were preformed by real-time PCR (TaqMan), and TREC-level was detected according to the number of CD3-positive cells. 5 case of ALL-CR and 17 normal individuals were served as controls. The results showed a dramatic reduction of TREC values in all groups of patients. The mean value of TRECs was 0.53 +/- 1.52 copies/1000 PBMNC and 2.01 +/- 3.93 copies/1000 CD3+ cells in adult B-ALL (p = 0.0005, p = 0.0123), 0.11 +/- 0.15 copies/1000 PBMNC and 0.23 +/- 0.27 copies/1000 CD3+ cells in B-CLL (p = 0.0015, p = 0.0381), 0.71 +/- 1.34 copies/1000 PBMNC in B-NHL (p = 0.0017), 0.53 +/- 0.90 copies/1000 PBMNC in MM patients (p = 0.0018), as compared with 3.76 +/- 3.42 copies/1000 PBMNC and 5.87 +/- 4.96 copies/1000 CD3+ cells in normal individuals, the TREC level was significantly decreased in all groups of B-cell lymphocytic malignancy, as well as in ALL-CR group. However, the TREC level in childhood B-ALL was significant higher than those in adult B-ALL group. It is concluded that the function of thymic recent outputting naive T cells in B-cell malignancies significantly decreases, however, the individual difference of thymic output function is obvious. The thymic recent output function can not be recovered during CR phase in patients with B-cell malignancies, so that dynamic analysis of TREC level is necessary.

Expressions of transcription factor GATA-1 and GATA-2 genes in bone marrow stromal cells from patients with leukemia / 中国实验血液学杂志

In order to investigate expressions of transcription factor GATA-1 and GATA-2 genes in the bone marrow stromal cells (BMSCs) from patients with leukemia or normal controls, bone marrow stromal cells from 34 normal cases and 42 cases with leukemia were cultured long-term in vitro. Nonadherent cells (bone marrow hematopoietic cells) and amplified adherent cells (BMSC) were collected separately. Expressions of GATA-1 and GATA-2 genes were analyzed by using RT-PCR-ELISA; the semi-quantitative expression levels of GATA genes in the BMSCs from patients with leukemia were compared with normal controls. The results showed that expressions of GATA-1 and GATA-2 genes could be detected in the BMSCs and the bone marrow hematopoietic cells from both normal controls and the cases of leukemia. The expression ratio of GATA-1 in the BMSCs from acute lymphocytic leukemia (ALL) (85.7%) was similar to the normal controls (88.2%), whereas the expression ratios in BMSCs from acute myelocytic leukemia (AML) (55.6%) and chronic myelocytic leukemia (CML) (41.2%) were significant lower than the normal controls (P < 0.05). The rank of expression level of GATA-1 gene in the BMSCs was "ALL>AML>normal>CML". There was no difference in the expression level of GATA-2 gene within the BMSCs from normal controls and patients with leukemia. The ranks of expression levels of GATA-1 and GATA-2 genes in bone marrow hematopoietic cells were "AML>normal>ALL>CML" and "AML>CML>ALL>normal". The dominant expression of GATA-2 gene was found in the BMSCs from AML, CML or normal controls. It is inferred that the expressions of GATA-1 and GATA-2 genes in the BMSCs of normal controls and patients with leukemia may influence the regulation of hematopoiesis in the bone marrow stroma and it is worthy of further study to explore their roles in pathogenesis and development of leukemia.

Peripheral blood naive T cell level and its T cell receptor Vbeta repertoire usage profile in patients with chronic myelogenous leukemia / 中华血液学杂志

<p><b>OBJECTIVE</b>To analyze peripheral blood naive T cell level, its T cell receptor (TCR) Vbeta repertoire usage profile and clonality for evaluating the recent thymic output function and the expansion feature of TCR Vbeta subfamily T cells in patients with chronic myelogenous leukemia (CML).</p><p><b>METHODS</b>Quantitative detection of T-cell receptor excision DNA circles (TRECs) in peripheral blood mononuclear cells (PBMNC) from 20 cases of CML was preformed by real-time PCR (TaqMan) analysis, and TRECs-number in T-cells was calculated from peripheral blood CD3-positive cell rate. The expression and clonality analysis were detected by RT-PCR and Genescan technique in PBMNC from 14 out of the 20 patients. Nine normal individuals served as controls.</p><p><b>RESULTS</b>A dramatic reduction of TRECs value in patients with CML was detected as compared with that in normal controls. The mean value of TRECs was 0.06 +/- 0.16 copy/1000 CD3(+) cells in CML patients while 6.84 +/- 4.71 copies/1000 CD3(+) cells in normal controls (P < 0.01). The 1 - 12 Vbeta subfamilies were variably expressed in samples from 14 patients. Genescan analysis identified clonal expanded T cells of some Vbeta subfamily from 13 cases. Vbeta3, Vbeta10, Vbeta19, Vbeta21 and Vbeta22 subfamilies clonal T cells were more frequently seen.</p><p><b>CONCLUSION</b>There is a prominent reduction of recent thymic output naive T cells function in CML. The predominant usage and clonal expansion of TCR Vbeta subfamily T cells could be identified, indicating that CML patients have specific immune response to leukemia associated antigen, in spite of their T cell immunodeficiency.</p>

Expression of SCL gene in bone marrow stromal cells from patients with leukemia / 中国实验血液学杂志

In order to investigate expression of SCL (stem cell leukemia) gene in bone marrow stromal cells (BMSC) and bone marrow cells from patients with leukemia and normal individuals, bone marrow mononuclear cells from AML (18 cases), CML (17 cases), ALL (7 cases) and normal individuals (33 cases) were cultured long-term in vitro. Nonadherent cells (hematopoietic cells) and amplified adherent cells (BMSC) were collected respectively. RT-PCR-ELISA assay was then performed to detect expression of SCL gene. The expression ratio of SCL gene were analyzed and its expression level was normalized by beta(2)M gene acting as an internal reference for the purpose of semi-quantitative analysis. The results indicated that the expression ratio of SCL gene was lower in BMSC from AML (27.8%) and CML (11.8%) than that in normal controls (69.7%, P < 0.05), while was higher in the nonadherent cells from CML (64.3%) than that in its corresponding BMSC (P < 0.05). Semi-quantitative analysis showed that SCL gene expression level in nonadherent cells from AML was higher than that in its corresponding BMSC (P < 0.05). In conclusion, the low-level expression state of SCL gene in BMSC from patients with AML and CML may be involved in the abnormal regulation of hematopoiesis in myelocytic leukemia.

Clonal expansion of TCR Vbeta subfamily T cells induced by bcr3-abl2 peptide / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the clonal expansion of T cell receptor (TCR) Vbeta subfamily T cells from cord blood induced by bcr3-abl2 peptide in vitro.</p><p><b>METHODS</b>T cells from 3 units of cord blood were amplified by anti-CD(3) monoclonal antibody (McAb) and IL-2 with or without synthetic b3a2 peptide. T cell specific cytotoxicity was analyzed by lactate dehydrogenase (LDH) assay, TCR Vbeta subfamilies by using reverse transcriptase-polymerase chain reaction (RT-PCR) and genescan technique.</p><p><b>RESULTS</b>bcr3-abl2 peptide specific cytotoxicity T cells were successfully induced from the 3 units of cord blood by synthetic b3a2 peptide. Compared with that in CD(3) McAb induced cells, distribution pattern of TCR Vbeta repertoire was different in T cells induced with b3a2 peptide. Oligoclonal and oligoclonal tendency TCR Vbeta subfamily T cells could be identified in cord blood T cells induced by b3a2 peptide in 1 or 2 weeks, whereas those induced by anti-CD(3) McAb and IL-2 were mostly polyclonal.</p><p><b>CONCLUSION</b>The cytotoxicity T cells with anti-CML specificity could be induced by b3a2 peptide. The specific anti-CML cytotoxicity may be derived from the clonal expansion TCR Vbeta subfamily T cells.</p>