Objective To construct a bicistronic retrovirus expression vector to coexpress Delta1 and EGFP genes. Methods Delta1-IRES-EGFP bicistronic retrovirus vector was constructed by linking full-length Delta1 cDNA with IRES-EGFP retrovirus vector. Transfected with the retrovirus vector by electroporation, the ecotropic- and amphotropic-packaging cell lines GP+E86 and PA317 were selected by addition of puromycin. Positive cell clones were obtained and tested for the efficiency of the retroviral supernatants. Two positive cell clones with relatively highest titers were picked up and used for ping-pong amplification assay to improve the retroviral supernatants titer. At day 2 after transfection, GFP expression in NIH3T3 was detected by FACS, and Delta1 expression was determined by RT-PCR assay. Transfected or wild type NIH3T3 were co-cultured with myoblasts cell line C2C12 to test whether the exogenous Delta1 protein was functional or not. Results The bicistronic retrovirus expression vector coexpressing Delta1 and EGFP genes was successfully constructed and verified by PCR and DNA sequencing. A high-level expression of functional Delta protein was detected in transfected NIH3T3 cells. Conclusion Retrovirus is a safe, highly efficient gene transfer system.

Vectors used to carry foreign genes play an important role in gene therapy, among which, the adeno-associated virus (AAV) has many advantages, such as nonpathogenicity, low immunogenicity, stable and long-term expression and multiple-tissue-type infection, etc. These advantages have made AAV one of the most potential vectors in gene therapy, and widely used in many clinical researches, for example, Parkinson's disease. This paper introduces the biological characteristics of AAV and the latest research progress of AAV carrying neurotrophic factor, dopamine synthesis related enzymes and glutamic acid decarboxylase gene in the gene therapy of Parkinson's disease.

Objective:To analyze the therapeutic effect and safety of finasteride on aged patients with isolated systolic hypertension (ISH)complicated benign prostatic hyperplasia (BPH).Methods:A total of 84 aged ISH + BPH pa-tients admitted from Jun 2011 to May 2013 were selected.According to medication plan,they were divided into fin-asteride group (n=44)and routine treatment group (n=40,only received routine treatment because they cannot use finasteride).Course of treatment was all six months.International prostate symptom score (IPSS),quality of life (QOL) questionnaire score,prostate-specific antigen (PSA),incidence rate of acute urinary retention,maximum u-rinary flow rate (Qmax),postvoid residual urine (PRV),prostate volume (PV)and incidence rate of adverse reac-tions were compared between two groups.Results:Compared with routine treatment group,there were significant reductions in IPSS [(13.2±2.4)scores vs.(9.4±1.1)scores],PRV [(21.4±4.6)ml vs.(9.0±2.3)ml],PV [(36.8±3.8)ml vs.(29.5±3.0)ml],PSA [(2.45±0.74)μg/L vs.(1.26±0.48)μg/L]and incidence rate of acute urinary retention (17.50% vs.9.09%),P<0.05 or <0.01,and significant increase in Qmax [(10.2±2.4) ml vs.(13.7±3.8)ml]and QOL score [(3.1±1.0)scores vs.(4.7±1.5)scores]in finasteride group (P<0.05). There were no significant difference in incidence rates of adverse reactions between two groups,P>0.05 all.Con-clusion:Therapeutic effect of finasteride is significant and adverse reactions don’ t increase significantly in aged pa-tients with isolated systolic hypertension complicated benign prostatic hyperplasia.

Objective To study the effects of chloroquine on proliferation and cell cycle of hepatocellular carcinoma cell line HepG2 cultured in vitro and provide basis for research on chloroquine in therapy for liver cancer.Methods HepG2 cells were divided into six groups:control group and chloroquine (8.00,16.00,32.00,64.00 and 128.00 mmol?L-1 )groups.The cell viability was determined by MTT,the cell cycle and apoptosis were detected by flow cytometry.Results Compared with control group,the cell viabilities were inhibited significantly 24 h after treated with chloroquine (32.00-128.00 mmol?L-1) or 48-72 h after treated with chloroquine (8.00-128.00 mmol?L-1) ( P

Objective To compare T2* value in healthy and diseased achilles tendons (AT) using 3D-ultra-short echo time (UTE),and to analyze the correlation between T2* value and clinical scores.Methods Fifteen patients (patients group) and ten volunteers (control group) were included and underwent an MR scan with UTE sequence.The T2* value of four ROI in AT were calculated.Independent sample t-test was used to compare the differences of T2* value between two groups.The correlation of T2* values with the American Orthopaedic Foot and ankle society ankle hindfoot scale (AOFAS) and achilles tendon total rupture score (ATRS) were analyzed.Results The T2* value of four ROI were statistically significantly higher in patients group than those of control group.There were negative correlation between T2* and AOFAS or ATRS scores (r=-0.92,-0.90,both P＜0.01).Conclusion The T2* value of pathologic AT increases.The quantitative MRI UTE-T2* can evaluate AT from the biochemical structure and correlates with clinical outcomes,which is helpful to diagnose AT in the early stage.

Objective To investigate the effect of 5-azacytidine on cardiomyogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Method BMSCs were isolated from the marrow of adult SD rat’s femoral /tibial bones.Different concentration of 5-azacytidine were added to primary BMSCs on 3 d and cultured for different times.Cardiomyogenic differentiation of BMSCs was observed by immunohistochemistry and RT-PCR. Result After treated with 5-azacytidine, BMSCs proliferated slowly, became spindle-shaped after 10 d and aligned in a striated pattern after 20 d.TnT positive cells were showed by Immunohistochemistry and they expressed two cardiac-marked genes GATA-4 and ?-MHC.Thus 5-azacytidine induced cardiomyogenic differentiation of BMSCs in a time and concentration-dependent manner. Conclusion Our study suggests that bone marrow mesenchymal stem cells can differentiate into cardiomyocytes in vitro. They are ideal donor cells in cellular cardiomyoplasty for treatment of myocardial infarction.

Objective To study the producing method for Euphorbia pekinensis promoted by endophytic fungi.Methods The tissue culture plantlets were innoculated with endophytic fungi,then it was transplanted to the pods and cultured for one year.The biomass and content of terpenoids(isoeuphpekinensin and euphol)were determined.Results The growth of the E.pekinensis was promoted by endophytic fungi E4(Fusarium sp.Ⅰ)and E5(Fusarium sp.Ⅱ).The fresh weight of the root increased 49.45% and 59.74% for E4 and E5 treated,respectively.The plants dry weight coefficient increased 2.99% and 6.48% for E4 and E5 treated,respectively.The contents of isoeuphpekinensin and euphol in the plants were increased for tissue cultured in both bottles and pods.For the plants cultured in pods for one year,the isoeuphpekinensin and euphol increased 92.79% and 40%,respectively treated by E4.The isoeuphpekinensin and euphol increassed 105.32% and 241.38%,respectively treated by E5.Conclusion The endophytic fungi E4 and E5 could promote the growth of E.pekinensis and accumulated terpenoids.

BACKGROUND: There have been reports about the cochlea injury after high-velocity projectile wounding. The effect on ultrastructure of cochlea after blast was still unclear.OBJECTIVE: To investigate ultrastructural changes of cochlea and cochlear nerve after maxillofacial blast wound.DESIGN:A randomized controlled observational trail with dogs as subjects.SETTING: Otolaryngeal Department and Maxillofacial Surgery Laboratory of the Third Military Medical University of Chinese PLA.MATERIALS: The study was conducted from August 1995 through July 1997. The animal model in maxillofacial blast wound was established in Animal Center of Xinqiao Hospital of the Third Military Medical University of Chinese PLA. Specimens were treated in Maxillofacial Laboratory and observed in Electron Microscope Laboratory. Totally 15 dogs of either sex (weighting 9. 5 - 13.5 kg, mean 11.3 kg ) were randomly divided into three groups with 5 in each group. Two groups served as trauma groups 1 and 2 and the other group as control.METHODS: The maxillary and mandibular regions of 10 dogs in trauma groups were wounded by model 8 cardboard-shelled detonators to establish animal model of maxillofacial blast wound. At the 1st and 6th hour after trauma the wounds were examined and specimens of cochlea and cochlear nerve were dissected out for electron microscopic observation to study the ultrastructural changes. The specimens in the control group were treated the same way as those in the trauma groups except for blast injury.RESULTS: After wounded, the cochlea and cochlear nerve in the early period manifested cilia disorder, edema of the nerve and mitochondrial degeneration. At the 6th hour after trauma there were extensive degeneration in cochlea and cochlear nerve, cilia fallen off hair cells and dissolution of the structures in nerve sheath.CONCLUSION: The ultrastructural changes of cochlea and cochlear nerve are severed as a result of maxillofacial blast wound, but in early period the injury is reversible. So it is very important that early cure should be emphasized in treatment.

p21 WAF1/CIP1 gene is known for a most important cell cycle regulator as well as its roles in apoptosis and differentiation. This review focuses on p21 WAF1/CIP1 gene functions and its association with carcinogenesis. Better understanding of the structure and function of p21 WAF1/CIP1 gene may help to comprehend molecular mechanisms of cancers and to facilitate diagnosis and treatment of malignancy.

OBJECTIVE:To provide reference for developing the implementation path of patent standardization for pharmaceuti-cal industry. METHODS:For the situation that pharmaceutical industry is still in the exploratory stage and lacks of viable experi-ence,the mature experiences of electronic communication industry in patent leading standardization implementation,authoritative organizations helping implementing patent standardization and industry alliance to promote the development of patent standardiza-tion were used as reference,combining with standard development cycle,trying to develop the implementation path of patent stan-dardization that met characteristics of pharmaceutical industry. RESULTS & CONCLUSIONS:The implementation path of patent standardization for pharmaceutical industry was divided into standard formation stage,promotion stage and upgrade stage. The main path in each stage was from patent to registration standards officially formed,guiding by policies and regulations of Food and Drug Administration to conduct standard promote by drug generation,and upgrading the drug standard by patent replacement.