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 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.

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 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.

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.

Objective To analyze causes of complication of long bone fracture of inferior extremity treated with interlocking intramedullary nail,and put forward prophylaxis and treatment measures.Methods Retrospective research on 140 patients of long bone fracture of inferior extremity treated with interlocking intramedullary nail from Oct.2002 to Oct.2005,to find out post-operative complications and put forward prophylaxis and treatment measures.Results All fractures were bone union,and complications occurd in 25 patients.Conclusion Interlocking intramedullary nail is preferred surgery to long bone fracture of inferior extremity.

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.

Objective To investigate the clinical and X-ray characteristic findings of the cystic and expansive bone metastasis.Methods The clinical and X-ray findings of the cystic and expansive bone metastasis in 35 cases were analysed with the literaturereviewed.There were 21 males and 14 females,the age ranged from 35 to 70 years with average of 52 years.Results The bone metastasis was found 1~10 years after primary tumor diagnosed in 29 cases befor primary tumor diagnosed in 5 cases and the primary tumor did not know in one case.The primary neoplasmes included nasopharyngeal carcinoma in 10,lung carcinoma in 9,renal carcinoma in 5,breastcarcinoma in 4,thyroid carcinoma in 3,hepatic carcinoma,prostatic carcinoma,esophageal carcinoma and primary focus unknown in onerespectively.The metastatic lesions localized in the long bone in 20 cases,ribs in 4,pelvis in 4,vertebrae body in 2,scopula in 2,clavicle,metacarpal and skull in one respectively.There were 16 cystic destruction,9 expansive destruction,10 both cystic and expansive.The marginated lesions were in 27 and 8 unclear in outline of lesion was in 8.There were distruction in the bony cortex with soft tissue in 7,and sclerosis in outline of lesion in 11 and bone crest and spot clacification inside the lesions in 6.Conclusion The time of metastases occured of cystic and expansive bone metastases is depended on the differentiating degree of primary tumor cell its developmental speed.

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.