Research Advances in the Mechanisms of BRD4 and Its Inhibitors in Hematologic Malignancies--Review / 中国实验血液学杂志

Bromodomain-containing protein 4 (BRD4) is one of the most important members in the bromodomain and extra terminal domain(BET) family, it plays an important role in cellular physiology in human body, such as cell cycles, cell proliferation, and immune response. Recent studies have shown that BRD4 is associated with occurrence and development of acute myeloblastic leukemia, multiple myeloma and lymphoma. The mechanisms of BRD4 in hematologic malignancies including the regulation of c-Myc expression, and participation of the composition of super-enhancer, etc. At present, many kinds of inhibitors have been developed to target inhibit BRD4 for therapy in hematologic malignancies, and some of BRD4 inhibitors have entered phase Ⅱ clinical trials, which suggested that BRD4 inhibitors are expected to become new therapeutic agents for hematologic malignancies. In this review, the research advance of BRD4 and BRD4 inhibitors in hematologic malignancies was summarized briefly.

The Genome-Wide Changes in Expression Profile of CML T Cells After Up-regulation of TCRζ Chain Expression / 中国实验血液学杂志

OBJECTIVE@#To analyze the changes in the gene expression profile of T cells in CML patients after TCRζ up-regulation expression, and to explore the molecular mechanism of T cell reactivation after transgenic up-regulation of TCRζ.@*METHODS@#The peripheral blood mononuclear cells(PBMCs) from 3 newly untreated chronic-stage CML patients were collected, and the CD3@*RESULTS@#A total of 2248 differentially-expressed genes were obtained, including 553 up-regulated genes and 1695 down-regulated genes in experimental group as compared with those in control group (P<0.05) . The GO and KEGG enrichment analyses showed that differentially expressed genes involved in the biological processes related to T cell immune function, such as TCR signaling pathway, T cell proliferation and activation. Some of core genes involved in promoting the TCR signaling pathway, T cell proliferation, activation and apoptosis pathways were significantly up-regulated, while some core genes involved in inhibiting T cell activation were significantly down-regulated.@*CONCLUSION@#The molecular mechanism of the significantly improved T cell activation and proliferation ability in CML patients after TCRζ up-regulation may be related to the differential transcripts mediated signaling pathways of T cell activation, proliferation and apoptosis.

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.

Immune Tolerance Mediated by TIM-3,LAG-3 and BTLA and Their Reversion in Hematological Malignancies-Review / 中国实验血液学杂志

The main mechanism of tumor immune suppression is due to the T cell exhaustion which is mediated by abnormal expression of T-cell immunosuppressive receptors in immune cells. Blocking these molecules may restore partial or all functions of T cells. This article reviews the advance on the role of the newly discovered T cell immuno-suppressive receptors such as TIM-3, LAG-3 and BTLA, including their mediated T cell-immune tolerance and the study of targeted immunotherapy in hematological malignancies, so as to provide the new strategy for immune-targeted therapy for hematological malignancies.

Effects of Down-regulating PPP2R5C Expression on Expression Profile of TAL1-related Regulating Genes in Jurkat Cells / 中国实验血液学杂志

<p><b>OBJECTIVE</b>Based on our previous study showing the inhibition of lenkemia T cell proliferation by down-regulating PPP2R5C expression, this study was aimed to analyze the influence of down-regulating PPP2R5 expression via RNA interference on genes relatied with TAL1 signaling pathway by using gene chip technique.</p><p><b>METHODS</b>The PPP2R5C-siRNA799 was transduced into Jurkat cells by nucleofection, the total RNA was isolated from treated Jurkat cells after culture for 48 hours; the target sequences were prepared by revevse transcription after mRNA purification, and were hybridized with affymetrix gene expression profile chip 3' IVT. The original image data were collected using affymetrix gene chip scanner 3 000, and the gene expression profile was analyzed using gene spring GX 11.0 soflware.</p><p><b>RESULTS</b>The expression of all 26 genes related with TAL1 signaling pathway was changed, out of which the expression of 15 genes were up-regulated and the expression of 11 genes was down-regulated in PPP2R5C-siRNA 799-transfected Jurkat cells. The genes with significantly up-regulated expression were GATA1, TCF4, XRCC6 and TCF3, while the genes with significantly down-regulated expression were SIN3A and RUNX1.</p><p><b>CONCLUSION</b>The down-regulation of PPP2R5C gene expression in Jurkat cells via RNA interference to a certain degree can inhibit TAL1 signaling pathway genes, thereby suppresses the proliferation of Jurkat cells.</p>

Overexpression of eIF4E gene in acute myeloid leukemia and its relation with disease progression / 中国实验血液学杂志

The aim of this study was to detect the expression level of eIF4E gene in patients with non-treated, remission and non-remission/relapse acute myeloid leukemia (AML), and other non-malignant haematologic diseases so as to analyze and reveal the relationship of eIF4E gene expression with AML progression. SYBR Green I RT-PCR was used to assay the expression level of eIF4E mRNA extracted from bone marrow mononuclear cells in 30 patients with AML (6 in M2, 5 in M3, 8 in M4, 10 in M5, 1 in M6) and 20 patients with non-malignant hematologic diseases. The β2-microglubin(β2M) was used as internal reference and the formula 2(-ΔCt)×100% was applied to calculate the expression level of eIF4E gene. The results showed that the eIF4E expression level (7.098 ± 5.544)% in patients with non-treated and non-remitted/relapsed AML was significantly higher than that in patients with remission (0.964 ± 0.312)% (P < 0.01) and non-malignant hematologic diseases (0.248 ± 0.163)% (P < 0.01). There was no difference between latter two group patients, even though the expression level of eIF4E gene in patients with M4 and M5 was higher. As compared with non-malignant hematologic diseases, the expression level of eIF4E gene of patients with remission patients showed no significant difference. It is concluded that the over-expression of eIF4E gene has been found in patients with AML, and its level obviously decreases along with remission of disease, thus the eIF4E gene may be a surveillance parameter for disease progression.

One-year continuous observation of change in peripheral T cell subsets in workers exposed to low levels of benzene / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To observe the T cell subsets and blood cells in the peripheral blood of workers exposed to low levels of benzene for one year, and to investigate the relationship between T cell function impairment and benzene-induced hematopoietic injury after benzene exposure.</p><p><b>METHODS</b>Eighty-eight workers (58 males and 30 females, aged 18 ∼ 22 years) who just began to work in the workshop of a paint factory with exposure to benzene in Guangzhou, China were assigned to experimental group, and 88 workers (58 males and 30 females, aged 18 ∼ 25 years) who worked in the workshop without exposure to benzene were selected as controls. The blood samples of the workers were examined once every 4 months to measure the percentages of peripheral T cell subsets and peripheral blood cell counts in the one-year study. The benzene concentrations at operation points were also measured.</p><p><b>RESULTS</b>The peripheral blood cell counts in the benzene-exposed workers had no significant changes in the first and second examinations; the white blood cell (WBC) counts in the experimental group in the third and fourth examinations were significantly lower than that in the control group [(6.4 ± 3.0)×10(9)/L and (6.3 ± 2.7)×10(9)/L vs (7.3 ± 3.0)×10(9)/L, P < 0.05], and the platelet (PLT) count in the experimental group in the fourth examination was also significantly lower than that in the control group[(179 ± 74)×10(9)/L vs (189 ± 70)×10(9)/L, P < 0.05]. Compared with those in the control group (CD4+: 54.29 ± 12.78%, CD8+: 37.25 ± 12.30%), the percentage of CD3+ T cells in the experimental group increased in the third examination; the percentage of CD4+ T cells in the experimental group decreased continuously in the second, third, and fourth examinations (50.77 ± 11.05%, 45.40 ± 9.41%, and 41.27 ± 10.62%), while the percentage of CD8+ T cells in the experimental group kept increasing (46.07 ± 10.18%, 50.36 ± 10.62%, and 56.40 ± 9.41%) (P < 0.05).</p><p><b>CONCLUSION</b>The change in T cell subsets precedes that in the blood system in the workers exposed to low levels of benzene.</p>

Supportive effects of conditioned culture media of human umbilical cord mesenchymal stem cells on hematopoiesis in vitro / 中国实验血液学杂志

This study was aimed to explore whether the conditioned culture medium of human umbilical cord-derived mesenchymal stem cells (hUC-MSC) has supportive effects on hematopoiesis in vitro. hUC-MSC were cultured in 75 cm(2) culture flasks at a concentration of 2×10(6) cells per flask. After 48 h, the conditioned culture medium was harvested. CD34(+) cells were isolated with the human cord blood CD34 positive selection kit. The CD34(+) cells were plated in three different culture systems: the culture supernatant from hUC-MSC added into incomplete methylcellulose without recombinant human cytokines as conditioned culture medium; the complete methylcellulose medium with recombinant human cytokines as positive control medium; incomplete methylcellulose adding DMEM/F12 with 10% FBS instead of conditioned culture medium as the negative control medium. After 14 days of culture, colonies containing ≥ 50 cells were scored and types of colonies were classified under inverted microscope. The immunophenotypes of cells which were collected from the colonies were detected by flow cytometry. The results showed that conditioned culture medium of hUC-MSC supported the differentiation of CD34(+) cells into CFU-G (47.67 ± 0.58), CFU-GM (48.67 ± 4.73) and CFU-M (3.00 ± 2.00) in vitro, while the CFU-E, BFU-E or CFU-GEMM were absent. Comparatively, in the positive control medium all kinds of CFU were observed. Interestingly, the percentage of CD45(+)cells of CFU in conditioned culture medium (97.43 ± 2.15)% was more than CD45(+)cells in positive control medium (39.69 ± 0.96)% (P < 0.05). It is concluded that the conditioned culture medium of hUC-MSC has been confirmed to have ability to support hematopoiesis separately in vitro. Besides, it enhances the differentiation of CD34(+) cells into myeloid cells except cells of erythroid lineage.

Immune regulation role of A20 and its clinical significance / 中国实验血液学杂志

A20 was originally identified as a TNFα-induced protein 3 (TNFAIP3), a key regulator of inflammation signalling pathways, as well as a NF-κB inhibitor. It plays a critical role in regulation of innate and adoptive immunity. Recently, A20 has also been proposed to function as a tumor suppressor. Lacking A20 gene is involved in inflammation-mediated autoimmune disease and tumorigenesis in several human B-cell lymphomas. Current advance concerning the feature of A20 expression in immune cells, the biological function, the immune regulated function in native immunity, humoral and cellular immunity, the inactivation of A20 in lymphocytic malignancies and the polymorphism and abnormal expression of A20 in autoimmune disease indicate that the clinical significance of A20 should be worthy to recognize and to be further employed in induction of immune tolerance, antitumor immuno-regulated therapy and antiviral immunotherapy and so on.

Establishment of stable subline of K562 cells expressing human leucocyte antigen a1101 / 中国实验血液学杂志

The aim of this study was to establish a stable subline of K562 cells expressing the HLA-A(*)1101 protein, which was expected to provide target cells for characterizing the HLA-I restrictive antigen specific cytotoxic T lymphocyte (CTL) effects against chronic myeloid leukemia (CML). The HLA-A(*)1101 protein encoding gene was amplified from peripheral blood mononuclear cell (PBMNC) of CML patient by RT-PCR; the 2A peptide linker (D-V-E-X-N-P-G-P) gene was linked to the 3'terminal of the HLA-A(*)1101 gene by recombinant PCR, then the recombinant was cloned into the pEGFP-N3 plasmid which contains an enhanced green fluorescent protein gene, and the eukaryotic recombinant expression vector containing HLA-A(*)1101-T2A-EGFP transcription box was constructed; the pEGFP-N3 vector and recombinant vector was separately electroporated into K562 cells. The expression of GFP was monitored by fluorescence microscopy, finally stably transfected sublines of K562 cells containing HLA-A(*)1101 gene, and of K562 containing pEGFP-N3 vector were obtained by G418 selection; the transcriptional or translational expression of HLA-A(*)1101 gene was detected with RT-PCR and flow cytometry respectively. The results indicated that the eukaryotic expression vector HLA-A(*)1101-T2A-EGFP plasmid was successfully constructed; after G418 selection for 2 months, two sublines of K562 cells (HLA-A(*)1101(+)K562, pEGFP-N3(+)K562) expressing GFP were constructed. The expression of HLA-A*A1101 gene could be determined in HLA-A(*)1101(+)K562 cell line by RT-PCR, while the pEGFP-N3(+)K562 cells could not express HLA-A*A1101 gene. HLA-A(*)1101 protein and GFP double positive HLA-A(*)1101(+)K562 cells were up to 88.5%, which was obviously higher than pEGFP-N3(+)K562 cells (0.698%) by flow cytometric analysis. It is concluded that a simple and effective method to select HLA-A(*)1101(+)K562 cells has been established and a subline of K562 cell expressing HLA-A(*)1101 protein on its cell membrane was successfully constructed, which provides the tool cells for further studying the specific cellular immunity against-CML.

Expression of CD133 in the bone marrow of patients with myelodysplastic syndrome and its clinical significance / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the expression of CD133 in the bone marrow of patients with myelodysplastic syndrome (MDS) and explore its clinical significance.</p><p><b>METHODS</b>The expression of CD133 and CD34/CD38 in the bone marrow was detected using flow cytometry in 31 cases of refractory anemia with excess blasts (RAEB), 10 cases of refractory cytopenia with multilineage dysplasia (RCMD) and 11 cases of aplastic anemia (AA).</p><p><b>RESULTS</b>The percentage of CD133-expressing cells was 6.75% in patients with RAEB, significantly higher than that in patients with RCMD (1.41%) and AA (2.70%) (P<0.05); the percentage of CD133-positive cells were similar between the latter two patient groups (P>0.05). The percentage of CD34(+)/CD38- cells was similar in the 3 groups (P>0.05), all lower than 1%.</p><p><b>CONCLUSIONS</b>Advanced MDS patients are characterized by an increase of CD133-expressing cells, suggesting the value of CD133 in the diagnosis of RAEB. CD34(+)/CD38- cells do not show a significant value in the diagnosis of MDS.</p>

Abnormal increase in CD8(low) T lymphocyte in patients with occupational chronic lead poisoning / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To analyze the changes in CD8(low) T lymphocyte subsets in patients with occupational chronic lead poisoning.</p><p><b>METHODS</b>Flow cytometric analysis was used to count the numbers of CD8+ cells. 23 patients with occupational chronic lead poisoning and 20 controls were examined.</p><p><b>RESULTS</b>Compared with control group (8.21% ± 3.02%), the CD8(low) T lymphocyte (12.98% ± 5.62%) were significantly increased in patients with occupational chronic lead poisoning.</p><p><b>CONCLUSION</b>Although the ratio of CD+ T lymphocyte is normal, the CD8 level is significantly decreased. The increase of CD8(low) T lymphocyte may be an important phenomenon of immuno-injury induced by lead. CD8(low) T lymphocyte could be an new direction for research of lead immuno-toxicity.</p>

Clonal expanded T-cell repertoire in MDS / 中国实验血液学杂志

The myelodysplastic syndromes (MDS) comprises a heterogeneous group of clonal hematopoietic stem cell disorders, while immunological abnormalities are frequently observed in such patients. T-cell mediated suppression of hematopoietic precursors contributes to the cytopenia and is related to the initiation and development of myelodysplastic syndromes (MDS). In this review, current knowledge concerning the feature of T-cell clonal expansion in MDS based on the analysis of T-cell receptor repertoire is summarized, including following issues: autoimmunity in MDS patients associated with T-cell mediated suppression of hematopoiesis, characteristics of T-cell clonal expansion in MDS patients, change of T-cell repertoire and recent thymus output function in MDS patients, efficiency of immunosuppressive treatment associated with T-cell clonal expansion in MDS patients and significance of T-cell clonal analysis in clinical application.

Change of expression pattern of CD3 genes in peripheral blood T-cells from CML patients / 中国实验血液学杂志

Our previous finding showed that down-regulation of CD3ζ gene was detected in patients with chronic myeloid leukemia (CML). In order to further elucidate the feature of T cell immune status in the signal transduction in CML patients, the expression patterns of all 4 CD3 genes were characterized in peripheral blood of patients, the expression levels of CD3γ, δ, ε and ζ chain genes were detected by real time qPCR with SYBR Green I staining in peripheral blood mononuclear cells (PBMNCs) from 17 cases of de novo CML patients in chronic phase and 17 cases of healthy individuals, the ß₂-microglobulin gene was used as an internal reference, and the mRNA expression level of each CD3 gene was evaluated by the 2(-ΔCt) x 100% method. The results showed that the median expression levels of CD3γ, δ and ε genes (2.344%, 0.515% and 3.516%) in CML patients were not significantly different from healthy individuals (p = 0.072, p = 0.190, p = 0.615, respectively), while the expression level of CD3ζ gene in PBMNCs from CML patients (0.395%) was lower than that from healthy individuals (1.538%) (p < 0.001). The expression patterns of 4 CD3 genes in proper order were CD3ε > CD3γ > CD3δ > CD3ζ in CML group, in contrast, the expression patterns were presented as CD3γ > CD3ε > CD3ζ > CD3δ in healthy group. It is concluded that the present study characterized the expression pattern of CD3γ, δ, ε and ζ chain genes in CML patients, lower expression of CD3ζ is the feature of TCR signal transduction immunodeficiency and the expression patterns of 4 CD3 genes are changed in CML patients.

Structural feature and biological function of PPP2R5C gene / 中国实验血液学杂志

PPP2R5C is one of the members of regulatory subunits of protein phosphatase 2A (PP2A), which plays a critical role in cell proliferation, differentiation and transformation, based on its induction of dephosphorylation of P53 at various residues. Recently, it was characterized that the alteration of expression pattern of PPP2R5C is associated with cell malignant transformation, thus PPP2R5C was thought as a marker for progressive disease in B-CLL. In this article the gene structure and biological function of PPP2R5C as well as relation of PPP2R5C with genesis and development of cancer were discussed.

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>

Serum proteomics in patients with RAEB myelodysplastic syndromes / 南方医科大学学报

<p><b>OBJECTIVE</b>To screen the molecular markers for refractory anemia with excess blasts in transformation (RAEB) in myelodysplastic syndromes (MDS) by serum proteome profiling.</p><p><b>METHODS</b>The serum protein were isolated from patients with RAEB, acute myeloid leukemia or normal subjects by 2-dimensional electrophoresis (2-DE), and the electrophoresis gels were obtained to identify the differentially reacting protein spots. The replica gels of the differentially reacting proteins were analyzed to locate the matching protein spots, which were identified by peptide mass fingerprint based on matrix-assisted laser desorption/ionization time of-flight mass spectrometry (MALDI-TOF-MS) and database searching.</p><p><b>RESULTS</b>Seven differentially expressed proteins in RAEB were found by 2-DE. Of the 7 proteins, 4 were identified by MALDI-TOF-MS to have significantly differential expression in RAEB, including dipeptidyl peptidase (DPP/CD26), polymerase (DNA directed) kappa, PRO2044 and an albumin-like protein.</p><p><b>CONCLUSION</b>2-DE-based serum proteome profiling helps identify serum proteomic biomarkers related to MDS. DDP/CD26 has increased expression in the serum in RAEB subtype MDS, suggesting its possible role in advanced MDS.</p>

Significance of quantitative detection of bcr-abl mRNA in chronic myeloid leukemia patients after allogeneic hematopoietic stem cell transplant / 中国实验血液学杂志

The objective of this study was to analyze the level of bcr-abl mRNA in peripheral blood (PB) after allogeneic stem cell transplantation (allo-SCT) in chronic myeloid leukemia patients, providing a experimental basis for diagnosing early relapse. bcr-abl mRNA levels in 78 PB and bone marrow (BM) samples from 15 CML patients after allo-SCT were detected by using real-time quantitative PCR. The results indicated that levels of bcr-abl mRNA before transplantation were high (median 29.303%) and decreased greatly (median 0) at the first month after allo-SCT. During the first year after allo-SCT, the patterns of serial bcr-abl transcripts varied in number, but the overall bcr-abl transcript levels significantly decreased at 6 months after allo-SCT. Majority of patients with undetectable or very low levels of bcr-abl mRNA were monitored after 1 year following transplantation. The hematological features of BM and PB in all detected patients remained normal. PB and BM bcr-abl values were not different significantly and had the similar trend of changes. It is concluded that the bcr-abl mRNA levels in CML patients change greatly early after allograft. Serial monitoring measurements for bcr-abl mRNA contribute to understanding the trend of change and effect of transplantation, also can be a guidance for starting therapy. But detectable levels of bcr-abl mRNA during the first 6 months do not indicate relapse. Measurements of bcr-abl mRNA of PB may be more suitable for routine monitoring long-term disease status in CML after allo-HSCT.

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.