The biggest highlight in the 55th ASH annual meeting is a series of revolutionary progress in the field of diagnosis,pathogenesis,prediction,prognosis and minimal residue disease (MRD) of malignant blood diseases guiding with next generation of sequencing (NGS) techniques.This article will share the major contents of 7 papers using NGS techniques in the ASH annual meeting with domestic readers here.

Objective: To find out a better technique of auricular reconstruction with Medpor framework for children to obtain better appearance of the reconstructed ear.Methods: The reconstructed ear was implanted by Medpor framework,which was covered by temporal superfacial fascia flap with superfacial temporal arteria.Skin flap of leftover ear covered rear of reconstructed ear,full thick skin graft covered one of obverse side,and infra-leftover ear was modified as auricularlobule.Results: Postoperative follow-up was as long as 24 months.All 12 patients obtained better appearance of the reconstructed ears than those of the operation in earlier period.Conclusion: The modified auricular reconstruction would be a better choice for the children with congenital lack of auricle or hypoplasia of auricle.

Objective:To construct the pEGFP-C2-hKGF vector,a neukaryotic expression plasmid of hKGF gene in order to provide experimental material for the studies of hKGF gene function and hKGF gene therapy afterward.Methods:The hKGF gene was amplified by reverse transcriptage polymerase chain reaction(RT-PCR).Total RNA was extracted from human embryo lung fibroblast(HLF)and then cloned into pMD18-T vector.The recombinant plasmid were screened and extracted.The pEGFP-C2-hKGF vector,an eukaryotic expression plasmid of hKGF gene was constructed after the object hKGF cDNA fragment was inserted into pEGFP-C2 vector and the recombinant were screened,cloned and extracted.The recombinant plasmid vector was transferred into AEC II,and expression of hKGF was assayed by immunohistochemistry.The effect of AEC II proliferation was analyzed by cell count.Results:609bp DNA fragment containing hKGF gene and restriction sites were obtained by RT-PCR.After cloned by pMD18-T vector and certified by restriction analysis and sequencing,pEGFP-C2-hKGF vector was successfully constructed by means of recombinant DNA technology.pEGFP-C2-hKGF encoding the green fluorescent protein(GFP),capable of directing the expression of functional hKGF successfully.Furthermore,hKGF had the effect on AEC II proliferation.Conclusion:The efficient expression vector of hKGF gene is constructed,which can be used to establish the hKGF gene therapy vector.

Objective:To investigate the mechanism of hypertrophic scar formation in different age periods.Methods:The expression of type I and type Ⅲ collagen of hypertrophic scar in different age periods was measured with immunohistochemistry which was divided into two groups 1～19 years old and 20～50 years old,and the same measure was adopted in normal skin divided into four groups-fetal,1～19,20～50,and more than 60 years old .The quantities of type I and type Ⅲ collagen and the ratios were analyzed.Results:①The ratio of type Ⅰ/type Ⅲcollagen of normal skin had a guadually increasing tendency.②The ratio of type Ⅰ/type Ⅲ collagen of HTS was increasing compared with normal skin(NS),and relative quantities of type I collagen of HTS was significantly increasing.The ratio of type Ⅰ/type Ⅲcollagen of HTS in group of 1～19 year old was highter than that in the group of 20～50 year old .Conclusion:Normal structure of the skin is decided by quantities and ratio of type I and type Ⅲ collagen.The formation of hypertrophic scar in different age periods relates to maladjustment of the ratio of type I and type Ⅲ collagen.Collagen synthesis and regulation of its constitute are finished after about six months.They keep relatively stable within 6 month to 2～3 years.

Objective To investigate the relationship between CD + 4CD + 25 regulatory T cells (CD + 4 CD + 25 Tr)and unexplained recurrent spontaneous abortion (URSA) Methods The expression rate of CD + 4CD + 25 Tr in peripheral blood was determined by double label flow cytometric analysis The samples were taken from 20 normal non pregnancy women, 17 URSA women and 20 normal pregnancy women Results The expression frequency of CD + 4CD + 25 Tr cells was decreased significantly( P 0 05) Conclusion The results suggest that downregulation of CD + 4CD + 25 Tr may be associated with URSA

Acute myeloid leukemia (AML) is the major topic in American Society Hematology (ASH)Annual Meeting and Exposition. There were 887 scientific articles on AML in the 53rd ASH Annual Meeting and Exposition in 2011,in which many studies focused on clinics.Here is introduction of o.ptimized regimens for AML and elderly AML,as well as targeted therapy for AML presented in the meeting.

Acute myeloid leukemia (AML) is the most commom hematologic malignancy in adults.There are so many highlights in the 54th ASH annual meeting in the field of mechanism,diagnosis and treatment of AML.Especially many studies have identified genes close related to AML development,clones related to relapse and molecular marker for prognosis using next-generation sequencing (NGS).Minimal residual disease (MRD) assay has been used for evaluation of therapy efficacy and posttherapy prognostication in AML treatment.The tyrosine kinase inhibitor targeting FLT3-ITD in this meeting is the best highlight in the treatment of refractory and relapsed AML.

In this year,ASH annual meeting reported several new targeting therapeutic drugs for chronic lymphocytic leukemia (CLL) and a part of lymphoma.Those drugs focus on some key targets in the signal transductional pathway of B cells.Although these targeting drugs individually did not improve the complete remission rate of CLL,but several clinical trial results showed that the combination therapy of those drugs with other drugs significantly improved the efficacy of CLL,and prolonged the progression-free survival of CLL patients.Therefore,experts believe that combination therapy and sequential therapy will start a new era in the treatment of CLL.This paper will share only the latest advances in the treatment of CLL and parts of lymphoma with domestic readers.

BACKGROUND: Mesenchymal stem cells from bone marrow stroma are able to differentiate into multiple mesoderm-type cell lineages. Because of the ease of their isolation and their extensive differentiated potentiality, mesenchymal stem cells become potential resource of cell and gene therapy. OBJECTIVE: To summarize biological characteristics and application of bone marrow mesenchymal stem cells. RETRIEVAL STRATEGY:An online search of Pubmed was undertaken by using the keywords of "Bone marrow mesenchymal stem cells, gene therapy, transplant"to identify the relevant articles published in English from January 1999 to December 2006. At the same time,Wanfang database was undertaken to identify the relevant articles published between January 1999 and December 2006 with the key words of "Bone marrow mesenchymal stem cells, gene therapy, transplant" in Chinese.The data were selected primarily, and then quotations of each article were checked. Inclusive criterion:The articles related to the biological characteristics and application of bone marrow mesenchymal stem cells were included. Exclusive criteria:the articles with repetitive research or Meta analysis were excluded.Totally 95 relevant articles were selected and 78 of them met the inclusive criterion. The 45 excluded articles were of old or repetitive content. LITERATURE EVALUATION: Selected articles were on basic experiment or clinical study of the biological characteristics, cell transplantation and gene therapy of bone marrow mesenchymal stem cells. Totally 33 articles of the 78 articles met the inclusive criterion were included. Of them, 4 articles were on the isolation of bone marrow mesenchymal stem cells, 3 on surface marker of bone marrow mesenchymal stem cells, 6 on biological characteristics of bone marrow mesenchymal stem cells, 3 on immune response between bone marrow mesenchymal stem cell transplantation and host and 17 on the application of bone marrow mesenchymal stem cells on disease therapy. DATA SYNTHESIS: In the suitable condition, bone marrow mesenchymal stem cells can differentiate into multiple cells, such as bone, chondrocytes, fat, muscle, tendon, neuron-like cells, myocardial cells and stroma-cells that support hematopoietic stem cells. Bone marrow mesenchymal stem cells seem to be the cells of immunosuppression and low-down immunogenicity. It makes bone marrow mesenchymal stem cells used in systemic transplantation, local implantation, as a vehicle for genes in gene therapy protocols or to generate transplantable tissues and organs in tissue engineering protocols. The results of these initial trials are very encouraging in animal test and several clinical trials are successful in some patients. But the enough clinical trails and long-term safety of therapeutics based on bone mesenchymal stem cells are needed. CONCLUSION: Biological characteristics of bone marrow mesenchymal stem cells have been gradually recognized. It is hoped that using bone marrow mesenchymal stem cells in clinic will bring major advances in the therapy of some diseases.

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