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 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 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.

Abortion, Induced ; Adult ; Chorionic Villi ; blood supply ; Endarteritis ; pathology ; Female ; Humans ; Pregnancy ; Pregnancy Complications ; pathology ; Stillbirth

Adult ; Heat Stroke ; Humans ; Male ; Occupational Diseases

Adult ; Ethylene Dichlorides ; poisoning ; Humans ; Male ; Occupational Exposure

Recurrent spontaneous abortion (RSA), which affects 1% to 5% of women of reproductive age, is difficult to treat in the clinical setting. In the investigations of iramunopathogenesis, diagnosis and treatment of RSA since the late 1980s, it was found that RSA was associated with abnormal maternal local or systemic immune response, the pathogenesis of autoimmune RSA was mainly associated with antiphosphlipid antibody (APA), while that of alloimmune RSA was due to the disturbance of maternofetal immunological tolerance. Systemic etiological screening process and diagnosis systems of RSA with immune type were developed, and anticardiolipin (ACL) + β_2-GP1 combining multiple assay for effective diagnosis of RSA with immune type was initially established. According to dynamic monitoring clinical parameters before and during gestation, low-dose, short-course and individual immunosuppressive therapy and lymphocyte immunotherapy for RSA. With immune type were initiated. The outcomes of the offsprings of patients with RSA were followed up, and the safety and validity of the therapies were confirmed. The research achievement leads to great progress in the diagnosis and treatment of RSA in China.

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.

Objective To provide guide for clinical diagnosis,classification and treatment of chronic kidney disease-mineral and bone disorder(CKD-MBD).Methods A total of 56 patients with chronic renal insufficiency were divided into three different groups:CKD4 period,CKD5 period and long-term hemodialysis(over 5 years).Their serum calcium,phosphorus,iPTH,bAP,and TRACP levels,and the bone mineral density were examined.Results The incidences of CKD-MBD in the three groups were 36.4%,73.0%,and 87.5%,respectively.High-turnover bone disease accounted for 65.79%,and the low-turnover bone disease accounted for 34.21%.The bAP and TRACP activity was significantly increased in patients with high-turnover bone disease and decreased in those with low-turnover bone disease.Conclusion CKD-MBD is a common complication of patients with chronic renal failure,with the main type being high-turnover bone disease,but the low-turnover bone disease can not be ignored.TRACP and bAP are the sensitive indicators of osteoblasts and osteoclast activity,and they could be used to identify high or low turnover bone disease.TRACP and bAP,together with serum calcium,phosphorus,iPTH,bone mineral density changes,can be used for a comprehensive analysis of bone metabolic state,so as to guide the diagnosis,sub-typing,drug selection and comprehensive treatment of CKD-MBD.