With the further study on molecular pathogenesis of multiple myeloma (MM),lots of novel targeted therapies emerged into clinical trials,which improved the response rate and life quality of MM patients.In the 56th American Society of Hematology (ASH) annual meeting,a series of special reports about latest developments in targeted therapy for MM patients were published.Efficacy was improved significantly by these reagents,which may provide the new treatment strategies for MM patients.The advances in the novel targeted therapies will be summarized in this paper based on the new reports in the 56th ASH annual meeting.

Numerous targeted therapies emerged into clinical trials, which improved the response rate and life quality of multiple myeloma (MM) patients. Series latest developments at targeted therapy for MM patients were reported on 58th American Society of Hematology (ASH) Annual Meeting, especially the results of combination with these novel agents showed a major highlight of this meeting. The advances in the novel targeted and biological therapies will be summarized in this paper.

12-O-tetradecanoylphorbol-13-acetate (TPA) is an activator of protein kinase C (PKC)extracted from the croton oil, it is one of the most powerful inducers of differentiation as well as a powerful tumor promoter. It is used by the study of induction and differentiation generally, because of it can induce the differentiation of tumor cell and promote the lymphocyte transformation and secrete of lymphokine, meanwhile,it is able to induce many kinds of cell of leukemic cell strain like HL-60, HEL and so on, Some literature reported that TPA had the function of differentiation of primary leukemic cell. The article mainly explains the induction and differentiation of leukemic cell by phorbol ester as well as its mechanisms, which to provide the theory evidence for prevention and cure of leukemia.

The clinical application of biological immunotherapy such as chimeric antigen receptor T cells (CAR-T) and novel targeted therapy has explored a new therapy for multiple myeloma (MM) treatment. Targeting B-cell maturation antigen (BCMA), allogeneic CAR-T, antibody-drug conjugate (ADC) and bispecific antibody targeting BCMA have achieved remarkable efficacy and safety in many clinical studies. This article introduces the latest immunotherapy for MM at the 62nd American Society of Hematology (ASH) Annual Meeting.

Objective To promote the differentiation of Flk1+CD31-CD34-cells isolated from adult adipose tissues into pancreatic islet endocrine cells in vitro.Methods Flk1+CD31-CD34-cells were first cultured and plated in medium supplemented with B27,epidermal growth factor(EGF),and basic fibroblast growth factor(bFGF).Next,the culture medium was changed.The glucose concentration in the serum-free medium was increased.At the same time,betacellulin and nicotinamide were added.Reverse transcription polymerase chain reaction(RT-PCR) was used to detect the expression of nestin,ngn3,insulin promoter factor-1(IPF-1),insulin,and glucagon before and after differentiation induction;immunofluorescent staining for nestin,insulin and glucagon and radioimmunoassay(RIA) for insulin.Results Initially,a nestin positive precursor cell population was found,then small round cells increased in number after 6 days.Later on,they were differentiated into islet-like clusters.The induced cells resulted in the formation of clusters which exhibited higher insulin secretion and other pancreatic endocrine hormones.RT-PCR detected an enhanced expression of pancreatic genes in the differentiated cells.Immunofluorescence revealed a high percentage of insulin-expressing cells in the clusters.Furthermore,the intra-cellular insulin content was detected by RIA after the induction culture.Conclusion These cells represent a previously unidentified adult intrinsic pancreatic precursor population and are a promising candidate for cell-based therapeutic strategies.

The present study retrospectively analyzed 2 patients suffering from refractory pure red cell aplasia after major ABO-incompatible hematopoietic stem cell transplantation who received treatment in the Henan Institute of Haematology between April 2004 and February 2006. Patient 1 was a 25-year-old female with acute lymphocytic leukemia in second remission, and patient 2 was a 16-year-old gid with acute myeloid leukaemia in second remission. The two patients received a transplant of human adipose tissue-dedved mesenchymal stem cells (1.0×106/kg). Both of them acquired rapid recovery from pure red cell aplasia without any side effects. These findings suggest that adipose tissue-dedved mesenchymal stem cells seem to be a promising therapeutic option in patients with refractory pure red cell aplasia after ABO-incompatible hematopoietic stem cell transplantation, in whom conventional treatment fails.

To assess the efficacy of cotransplantation of haploidentical mesenchymal stem calls (MSC) and hematopoietic stem cells (HSC) in the treatment of refractory severe aplastic anemia, Two child patients with refractory severe aplastic anemia admitted to Henan Institute of Haematology from August 2002 to December 2007 were selected. Adipose tissue-derived MSCs (AMSCs) were separately originated from haploidentical mother and peripheral blood stem calls (PBSCs) from HLA-identical sibling brother or sister of patients. The patient 1 received a cotransplantation of PBSCs and AMSCs (1 × 106/kg) at a dose of 4.5 × 108 mononuclear calls/kg (containing 4.41 × 106 CD34+ calls/kg and 0.11 ×105 CD3+ cells/kg); the patient 2 received a second PBSCT at a dose of 6.5 × 108mononuclear cells/kg (containing 4.62×106 CD34+ cells/kg and 0.12×105 CD3+ cells/kg) and AMSC (1 × 108/kg) from his haploidentical mother. The results show that the cotransplantation was successful. During the two years of follow up, the two patients exhibited good condition, with no other treatment or transfusion dependence.

BACKGROUND: There is no consistently effective therapy for patients with steroid-refractory acute graft-versus-host disease (GVHD). A variety of alternative approaches have been tested, including antithymocyte globulin, mycophenolate mofetil (MMF), pentostatin, and monoclonal antibodies; however, these treatments have been only modestly successful. OBJECTIVE: To further evaluate the efficacy of human adipose-derived stem cells (ASCs) as the salvage therapy for steroid-refractory acute GVHD. DESIGN: A clinical trial.SETTING: Department of Haematology, Henan Institute of Haematology, Henan Tumor Hospital.PARTICIPANTS: The clinical trial was performed at the Henan Institute of Haematology from September 2002 to August 2005. Eight patients were treated with ASCs for grades Ⅲ-Ⅳ steroid-resistant acute GVHD. The study was approved by the Ethics Committee at Henan Tumor Hospital and informed consent was obtained from patients and ASCs donors before they enrolled.METHODS: Eight patients with steroid-refractory grades Ⅲ-Ⅳ acute GVHD received intravenous infusions of ASCs. The ASCs dose was 1.0×106/kg. Seven patients were treated once and one patients twice. Four patients received ASCs from haplo-identical family donors and four from unrelated mismatched donors. MAIN OUTCOME MEASURES: The efficacy of human ASCs as the salvage therapy for steroid-refractory acute GVHD. RESULTS: No side effects were noted after the ASCs infusions. Acute GVHD disappeared completely in seven of eight patients and six of these seven patients are still alive after the median follow-up of 30 months (range 11-90 months) after the initiation of ASCs therapy. All four surviving patients were in good clinical condition and in remission of their hematological malignancy. Two patients died-one with no obvious response to ASCs died of multiorgan failure and one of relapse of leukemia. CONCLUSION: These results suggest that ASCs is a very promising treatment for severe steroid-resistant acute GVHD.

BACKGROUND: The treatment of chronic myelogenous leukemia (CML) is revolutionized by the tyrosine kinase inhibitor imatinib mesylate (imatinib). However, resistance to imatinib is increasingly recognized as a clinical problem, the prognosis of patients who develop imatinib resistance is poor, particularly in acute transformation phase of leukemia.OBJECTIVE: To characterize a novel CML cell line and to further elucidate the mechanisms of resistance to STI571.DESIGN: An observational comparative experiment.SETTING: Henan Institute of Haematology, Henan Tumor Hospital.MATERIALS: Thirty female BALB/c nu/nu mice with 5 weeks old were purchased from Animal Center, Chinese Academy of Medical Sciences. STI571 was kindly provided by Novartis (Nuremberg, Germany). VP-16 was purchased from Bristol-Myers Squibb (Munich, Germany); anti-P-gp from Santa Cruz Company, USA; anti-ab1 from BD Biosciences Company, USA. The disposal of experimental animal was coincidence with the ethical standard.METHODS: The experiment was performed in the Henan Institute of Haematology from September 2003 to November 2005. A novel K562 cell line (K562/VP16) was achieved after exposure of the K562 cells to VP16. A small subpopulation (SP K562/VP16) that was capable of excluding Hoechst 33342 in the K562/VP16 cell line was isolated by flowcytometry sorting. The rest of the K562/VP16 cells were classified as non-SP K562/VP16. In order to elucidate the mechanisms involved in K562/VP16 SP cells which became resistant to STI571, the expression of multidrug-resistant gene 1 (MDR1), Bcr-Abl and P-gp was detected in K562, non-SP K562/VP16, or K562/VP16 SP calls, respectively. Furthermore, one thousand cells of K562, K562/VP16 SP and non-SP cells were injected,respectively, intraperitoneally into the right flanks of ten 5-week-old female BALB/c nu/nu mice. The same experiment was repeated twice.MAIN OUTCOME MEASURES: Comparison of STI571 resistance and oncogenicity of non-SP K562/VP16 and K562/VP16 SP cells.RESULTS: The MDR-1 gene expression of the Mr 170 000 P-gp was detected in K562/VP16 non-SP and K562/VP16 SP cells but not in K562 cells. The expression levels of P-gp in the two K562/VP16 cell lines were similar (P ＞ 0.05).The levels of Bcr-Abl and Abl proteins were similar in the K562 cell line and in non-SP K562/VP16 and K562/VP16 SP cells (P ＞ 0.05). Compared with non-SP K562/VP16, the K562/VP16 SP cells were more resistant to STI571, and this resistance could hardly be reversed by many multidrug resistance inhibitors. In addition, in vivo study showed that the K562/VP16 SP cells induced oncogenicity in mice, while the K562/VP16 non-SP cells failed to do so.CONCLUSION: Bcr/abl gene amplification and MDR1 overexpression may not be an important clinical mechanism in the diversity of resistance development to imatinib treatment, and the development of drug resistance by leukemia cells may be at least partly due to a rare SP cells which drives leukemia occurrence and maintenance. So, these SP cells need to be targeted for effective cancer therapy.

Objective To explore the feasibility of establishment of NOD/SCID-mouse model with multiple myeloma by using plasma cells from myeloma patients.Methods The femurs and tibias were removed from the New Zealand white rabbits;the muscles,periosteum and cartilage tissues were cleared.Then each bone was cut into two pieces gently along its middle.The NOD/SCID mice weighing 25 - 30 g (4 - 6 weeks)were anesthetized by intraperitoneal injection;rabbit bone was inserted into the right side of the mouse back and engraftment of the bones was allowed to take place after 4 weeks.The 5000 000 purified plasma cells which expressed CD38 +/CD45 - were immunofluorescence labeled and then injected slowly into the implanted rabbit bone through the distal end.The mice were observed weekly;the plasma cells growth in mice was screened by the living-imaging system and the tumor from the mice was determined by biopsy.Results The implanted rabbit bone survived after 4 weeks.The tumor in mice was observed 2 weeks after the purified myeloma cells were injected into the rabbit bone,and it reached 100 mm3 after 8 weeks.Results of the living-imaging system showed that the myeloma cells had uptake in the rabbit bone after 2 weeks of injection and this phenomenon was more pronounced after 8 weeks of injection (2.4×10 4 vs .1.5× 10 5 ,P < 0.05 ).The tumor infiltrated with numerous plasma cells and osteoclasts increased upon the biopsy. Conclusion Rabbit bone marrow implanted into NOD/SCID mice can effectively support local injection of plasma cells of multiple myeloma patients,and the NOD/SCID-mouse model of myeloma has been established.This model can be used to study in vivo experiments related to myeloma and clinical therapeutic approaches for this disease.