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.

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.

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 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 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 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.

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 evaluate the effectiveness and safety of the combination of dexmedetomidine and propofol for sedation in pediatrics undergoing regional anesthesia.Methods Forty-eight children aged 2 -6 years old,scheduled for elective surgery under regional anesthesia,were randomly divided into the two groups.The dexme-detomidine plus propofol group(group DP,n=24)received an infusion over 10 minutes of dexmedetomidine 0.7μg/kg and propofol 1.0 to 2.5 mg/kg bolus for induction,then an infusion of dexmedetomidine 0.5μg·kg-1·h-1 and propofol 100-300μg·kg-1·min-1 for maintenance.The propofol group(group P,n=24)received a propofol 1.0 to 2.5mg/kg bolus for induction,then an infusion of propofol 150 -300μg·kg-1 ·min-1 for maintenance.If body movement was detected during surgery,a bolus of propofol 1 .0mg/kg was administered in both groups,and if there was another body movement within three minutes of the propofol bolus,a bolus of ketamine 1 .0mg/kg was adminis-tered.Hemodynamic data,respiratory rate,and oxygen saturation were recorded by researchers blinded to the study drugs.Recovery time and the primary outcome were evaluated by a modified Aldrete score.The occurrence of adverse events and the amount of supplemental ketamine used were also recorded.Results The consumption of propofol in the group DP was (149.00 ±24.93)μg·kg-1·min-1,and in the group P was (217.00 ±17.90)μg·kg-1· min-1.The propofol consumption was significant lower that in group DP(t=117.94,P=0.00).The needs for sup-plemental ketamine bolus in the group DP was 0.50(1.00)/case,and it was 2.00(1.00)/case in group P.The need for supplemental ketamine was less in group DP(Z=-4.48,P=0.00).Hypopnea occurred in 5 cases in group P, and 0 in group DP.The incidence of hypopnea was more in group P(χ2 =5.58,P=0.03).In group DP,the mean arterial pressure and heart rate at discharge were respectively (74.96 ±5.69)mmHg and (101.17 ±9.65)bpm,and their baseline values were (81.13 ±7.23)mmHg and (112.00 ±13.65)bpm.Both mean arterial pressure and heart rate in the group DP remained decreased at discharge(t=10.78,P=0.00;t=10.08,P=0.00;respectively).The discharge time of the group DP was (40.91 ±10.90)min and it was(39.67 ±14.95)min in group P.There was no significant difference in the discharge time(t=0.11,P=0.74).Conclusion The combination of dexmedetomidine and propofol can provide appropriate depth of sedation in pediatric patients undergoing regional anesthesia.

Objective To explore the relation between rheumatoid arthritis (RA) and the meteorological factors.Methods All RA cases in the group were followed for 12 months,patients were follow-up for every two weeks.In each followed-up,all RA cases were evaluated by the disease activity score-28 (DAS28) and the improved Jamieson questionnaire,and the correlation of the DAS28 and the average values of meteorolo-gical factors (temperature,humidity,atmospheric pressure,relative humidity,wind speed) by Pearson correlation coefficient.The moderate-severe activity cases were evaluated by chi-square test of different seasons.Results One hundred and eighty-eight cases completed the observation,One hundred and sixty-four cases were in moderate disease activities (x2=9.42,P=0.022).Sixty-eight cases were distributed in winter,which was significantly higher than other three seasons.The average temperature and wind speed was negatively related to the DAS28 (r=-0.743,P＜0.01; r=-0.576,P=0.003).Conclusion The activity of RA has the characteristics of seasonal changes,and is easily being aggravated in winter.Many factors may influence RA,including temperature and wind speed.Creating an appropriate micro-climate environment is beneficial for RA patients to reduce the disease activity.

The efficacy of different kinds of insulin analogue administered by multiple subcutaneous injection or continuous insulin infusion was compared. The two forms of administration all can well control blood glucose. Continuous subcutaneous injection has better compliance.