Antineoplastic Agents, Phytogenic ; therapeutic use ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; Phytotherapy ; Plants, Medicinal ; chemistry

In the last twenty years, using all-trans retinoic acid (ATRA) as a differentiation inducer, Shanghai Institute of Hematology has achieved an important breakthrough in the treatment of acute promyelocytic leukemia (APL), which realized the theory of reversing phenotype of cells and provided a successful model of differentiation therapy in cancers. Our group first discovered in the world the variant chromosome translocation t(11;17)(q23;q21) of APL, and cloned the PML-RAR alpha, PLZF-RAR alpha and NPM-RAR alpha fusion genes corresponding to the characterized chromosome translocations t(15;17); t(11;17) and t(5;17) in APL. Moreover, establishment of transgenic mice model of APL proved their effects on leukemogenesis. The ability of ATRA to modify the recruitment of nuclear receptor co-repressor with PML-RAR alpha but not PLZF-RAR alpha caused by the variant chromosome translocation elucidated the therapeutic mechanism of ATRA from the molecular level and provides new insight into transcription-modulating therapy. Since 1994, our group has successfully applied arsenic trioxide (As(2)O(3)) in treating relapsed APL patients, with the complete remission rate of 70% - 80%. The molecular mechanism study revealed that As(2)O(3) exerts a dose-dependent dual effect on APL. Low-dose As(2)O(3) induced partial differentiation of APL cells, while the higher dose induced apoptosis. As(2)O(3) binds ubiquitin like SUMO-1 through the lysine 160 of PML, resulting in the degradation of PML-RAR alpha. Taken together, ATRA and As(2)O(3) target the transcription factor PML-RAR alpha, the former by retinoic acid receptor and the latter by PML sumolization, both induce PML-RAR alpha degradation and APL cells differentiation and apoptosis. Because of the different acting pathways, ATRA and As(2)O(3) have no cross-resistance and can be used as combination therapy. Clinical trial in newly diagnosed APL patients showed that ATRA/As(2)O(3) in combination yields a longer disease-free survival time. With the median survival of 18 months, none of the 20 cases in combination treatment relapsed, whereas 7 relapsed in 37 cases in mono-treatment. This is the best clinical effect achieved in treating adult acute leukemia to this day, possibly making APL the first adult curable leukemia. Based on the great success of the pathogenetic gene target therapy in APL, this strategy may extend to other leukemias. Combination of Gleevec and arsenic agents in treating chronic myeloid leukemia has already make a figure both in clinical and laboratory research, aiming at counteracting the abnormal tyrosine kinase activity of ABL and the degradating BCR-ABL fusion protein. In acute myeloid leukemia M(2b), using new target therapy degradating AML1-ETO fusion protein and reducing the abnormal tyrosine kinase activity of c-kit will also lead to new therapeutic management in acute leukemias.
Antineoplastic Agents ; therapeutic use ; Benzamides ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Humans ; Imatinib Mesylate ; Leukemia, Promyelocytic, Acute ; drug therapy ; genetics ; metabolism ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Piperazines ; therapeutic use ; Protein-Tyrosine Kinases ; antagonists & inhibitors ; metabolism ; Pyrimidines ; therapeutic use ; Receptors, Retinoic Acid ; genetics ; metabolism ; Tretinoin ; therapeutic use

This article summarizes the progress of hematology in the recent tens years to show that experimental hematology used to pick up the 'hints' from clinical problems as the renewal of research directions and targets in experimental studies continuously. As the feedback, the results from lab investigations inserted into clinical practice and eventually made a quick modernization of hematology, which was actually a good model for the "translational research". The past few decades have witnessed tremendous advances in our understanding of normal hematopoiesis where genes dictate, epigenetics regulate, transcription factors mediate, and stem cells self-renew and differentiate. Dissection of disease pathogenesis not only elucidates molecular basis of disorders including hemoglobinopathy, aplastic anemia, hemophilia, hematopoietic malignancies such as leukemia and myeloproliferative disorders, but also provides therapeutic targets for drug development. Introduction of targeted therapies and combinatory targeting therapies greatly benefits hundreds of thousands of patients, and even turns acute promyelocytic leukemia from highly fatal to highly curable. In the 21st century the experimental hematology is entering the era of genomics and system biomedicine, and the pace of progress extrapolates to a prediction of hematologic neoplasms control in this century.
Animals ; Clinical Laboratory Techniques ; trends ; Hematologic Diseases ; genetics ; metabolism ; physiopathology ; Hematologic Neoplasms ; genetics ; metabolism ; physiopathology ; Hematology ; trends ; Humans

Objective: To observe the effect of acupuncture plus thunder-fire moxibustion on the expressions of matrix metalloproteinase-3 (MMP-3), tissue inhibitor of metalloproteinase-1 (TIMP-1) and transforming growth factor-β1 (TGF-β1) in cartilage of knee osteoarthritis (KOA) rats, and to explore the mechanism of acupuncture plus thunder-fire moxibustion in the treatment of KOA. Methods:Thirty Sprague-Dawley (SD) rats were randomly divided into a blank control group, a model group and an acupuncture-moxibustion group by random digits table, 10 rats in each group. Rats in the model group and the acupuncture-moxibustion group were injected with papain in the right posterior knee joint to prepare the models. The levels of MMP-3 and TIMP-1 in rat synovium of each group were measured by enzyme-linked immunosorbent assay (ELISA) after 2 weeks of treatment. The level of TGF-β1 was determined by Motic B5 Micro-camera system. Results:The levels of MMP-3 and TIMP-1 in the cartilage of the model group were significantly higher than those in the blank control group (allP<0.01); the levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were lower than those in the model group, and the between-group differences were statistically significant (all P<0.05). The levels of MMP-3 and TIMP-1 in the acupuncture-moxibustion group were higher than those in the blank control group, and the differences were statistically significant (all P<0.05). The level of TGF-β1 in cartilage tissues of the model group was significantly lower than that in the blank control group (P<0.01); the level of TGF-β1 in the acupuncture-moxibustion group was higher than that in the model group (P<0.05), but it was lower than that in the blank control group, and the between-group difference was statistically significant (P<0.05). Conclusion: Acupuncture plus thunder-fire moxibustion can effectively recover the abnormal expressions of MMP-3 and TIMP-1 in KOA model rats and somewhat up-regulate TGF-β1, which may be one of its mechanisms of acupuncture plus thunder-fire for KOA.

OBJECTIVETo identify genetic alterations in diffuse large B-cell lymphoma (DLBCL) and to analyse the relationship between the genetic aberrations and the clinical characteristics.

METHODSUsing comparative genomic hybridization (CGH) to investigate the genomic changes in 24 cases of DLBCL and to analyse the relationship between these aberrations and clinical parameters including Ann arbor stage, systemic symptoms, chemotherapy efficacy and survival.

RESULTSAberrations were detected in 62.5% patients of 24 cases; the most common chromosomal alterations included loss of 6q15-21 as well as gain of 18q11-ter, of which the incidences were 20.8% and 16.7%, respectively; with comparing clinical parameters between patients with normal CGH and abnormal CGH, we found that patients with abnormal CGH suffered more from stage III-IV and had higher incidence of systemic symptoms, poor chemotherapy efficacy and poor survival (P<0.05), but there was no difference observed in the incidence of extranodal involvement between two groups.

CONCLUSIONThe gains and/or losses of genomic DNA from DLBCL patients are the common molecular cytogenetic aberrations; loss of 6q15-21 and gain of 18q11-ter are nonrandom event to DLBCL patients; abnormal CGH is a clinical parameter reflecting malignant progressive course and poor survival to DLBCL patients.

Chromosome Aberrations ; Female ; Humans ; Karyotyping ; Lymphoma, B-Cell ; genetics ; pathology ; physiopathology ; Lymphoma, Large B-Cell, Diffuse ; genetics ; pathology ; physiopathology ; Male ; Nucleic Acid Hybridization ; Statistics as Topic

With recent development and progress achieved in the diagnosis and treatment of leukemia, relapse of disease still remains as the major problem in clinical management and the minimal residual disease (MRD) has been confirmed to be associated with leukemia relapse in the past decades. Due to the low sensitivity, morphology-based assays have limitation in the MRD monitoring. With the development of molecular assays, especially the real-time RT-PCR method have been a sensitive, precise and reliable tool to MRD detection. Numerous clinical studies demonstrated that the existence of MRD reflects the clinical and molecular response, and remain as a major prognostic factor for leukemia. Another important application of MRD detection is to evaluate the efficacy of different therapy at molecular level. In this paper, the different methods and their clinical application for MRD detection were systemically reviewed and it is confident that the establishment of standardized MRD detection system will be important in the clinical prevention for relapse of leukemia.
Flow Cytometry ; Humans ; Immunophenotyping ; methods ; Leukemia ; diagnosis ; genetics ; immunology ; Neoplasm, Residual ; diagnosis ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Human leukemia is closely associated with various genetic alterations such as chromosomal translocations and gene mutations. The use of retroviral transduction/bone marrow transplantation mouse model harboring these genetic abnormalities has been critical in understanding the molecular pathogenesis of leukemia and exploring new therapeutic target. Additional genetic events are verified to cooperate with fusion genes resulting from chromosomal translocations in acute myeloid leukemia (AML) to develop a leukemic phenotype in mice, such as C-KIT N822K with AML1-ETO, FLT3-ITD with PML-RARα, Meis1 with NUP98-HOX, and Cdx4 with MLL-AF9. Mouse model shows that BCR/ABL fusion gene induces chronic myeloid leukemia (CML), and suggests that GATA-2 L359V and high expression of Hes1 are key molecules in acute myeloid transformation of CML. Furthermore, combination therapy with Imatinib and arsenic sulfide for CML mice exerts more profound therapeutic effects than either drug as a single agent. This review focuses the recent progress and application of retroviral-mediated mouse models of myeloid leukemia, and discusses some factors influencing the mouse model establishment, including retroviral construction, retrovirus titer and hematopoietic microenvironment.
Animals ; Disease Models, Animal ; Leukemia, Myeloid ; genetics ; Mice ; Retroviridae ; genetics

This study is designed to serve as a reference for the establishment of health security systems for children’s critical diseases. Through analysis of the operation of Shanghai Children Hospital Care Aid (SCHCA), this study explored the financing model and management of a children’s critical disease healthcare system and analyzed the possibility of expanding this system to other areas. It is found that a premium as low as RMB 7 per capita per year under SCHCA can provide high-level security for children’s critical diseases. With the good experience in Shanghai and based on the current basic medical insurance system for urban residents and the new rural cooperative medical scheme (NRCMS), it is necessary and feasible to build a health security system for children’s critical diseases at the national level.
Adolescent ; Child ; Child Welfare ; Child, Preschool ; China ; Delivery of Health Care ; Health Policy ; economics ; legislation & jurisprudence ; Humans ; Infant ; Infant, Newborn

OBJECTIVETo characterize the structural and the functional feature of a novel gene HSPCSET isolated from human CD34+ hematopoietic stem/progenitor cells (HS/PCs).

METHODSBioinformatic technology was used to identify the structural features of the HSPCSET protein and perform the multiple sequence alignment. Yeast-two-hybrid system was used to identify the proteins interacting with the HSPCSET protein. After sequencing, we selected out the positive clones which had clear functions, and carried out beta-gal experiment and GST pull down assay to confirm the results. The cellular location of the HSPCSET was checked by immunofluorescence assay.

RESULTSThe HSPCSET protein belongs to a SET domain family, which is evolutionarily conserved across species. It implied that HSPCSET may have biologically important function. Using yeast-two-hybrid system, we showed that the protein sequence with SET domain might bind to 13 proteins, which involved in signaling transduction, transcriptional regulation, apoptosis, tumorigenesis, development, etc. And 4 proteins (GADD34, SIVA, DNAJ and PHF1) were confirmed by one-on-one back of the hybrid experiment, beta-gal test and GST pull down assay. When GADD34 and HSPCSET were co-transfected, they co-localized in the nucleus, suggesting a strong interaction.

CONCLUSIONThe novel gene HSPCSET is likely to have biologically important function. This study provides the basis for further studies of its function in hematopoiesis and tumorigenesis.

Amino Acid Sequence ; Animals ; Antigens, Differentiation ; metabolism ; Cell Cycle Proteins ; metabolism ; Computational Biology ; Conserved Sequence ; Hematopoietic Stem Cells ; metabolism ; Humans ; Molecular Sequence Data ; Protein Phosphatase 1 ; Protein Structure, Tertiary ; Proteins ; chemistry ; genetics ; metabolism ; Sequence Homology, Amino Acid ; Two-Hybrid System Techniques

To investigate the potential role and the mechanism of PLZF-RARalpha/RARalpha-PLZF double fusion gene in the pathogenesis of acute promyelocytic leukemia (APL) in vivo at systematic biological level, PLZF-RARalpha/RARalpha-PLZF double transgenic mouse model was established by intercross; the integration and expression of fusion genes were analyzed by PCR and RT-PCR; the disease phenotype was detected by morphological and pathological examination of peripheral blood and bone marrow cells, as well as flow cytometry assays; the effects of ATRA with or without tricostatin A on bone marrow blast cells from PLZF-RARalpha/RARalpha-PLZF double TM were observed. The results showed that leukemia occurred in 5 PLZF-RARalpha/RARalpha-PLZF double TM 7, 7, 9, 11 and 11 months respectively, out of them two (40%) with classic APL features, the others (60%) with chronic myeloid leukemia through an observation period of 18 months. The leukemia occurrence of PLZF-RARalpha/RARalpha-PLZF TM was about 10%, which was similar to PLZF-RARalpha TM as that reported before. The latency was over 6 months, not earlier than PLZF-RARalpha TM only. No morphologic changes of PLZF-RARalpha/RARalpha-PLZF double TM blast cells to ATRA were observed, but increased cytoplasmic-nuclear ratio and nuclear condensation in bone marrow blast cells were found in combination of ATRA with tricostatin A. It is concluded that PLZF-RARalpha/RARalpha-PLZF double fusion gene transgenic mice have heterogeneity of pathogenesis. HDAC inhibitors such as trichostatin A, in combination with ATRA, induce differentiation of the blast/promyelocytic cells from PLZF-RARa/RARa-PLZF double TM, but not ATRA alone.
Animals ; Antigens, CD34 ; blood ; Bone Marrow Cells ; drug effects ; immunology ; pathology ; Cell Differentiation ; drug effects ; Chorionic Gonadotropin ; genetics ; Disease Models, Animal ; Female ; Flow Cytometry ; Humans ; Hydroxamic Acids ; pharmacology ; Leukemia, Promyelocytic, Acute ; blood ; genetics ; pathology ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred CBA ; Mice, Transgenic ; Oncogene Proteins, Fusion ; genetics ; Pedigree ; Receptors, Chemokine ; blood ; Tretinoin ; pharmacology