AIM: To construct recombinant adenoviral vector carrying the LEDGFp52 gene by homologous recombination in bacteria and to detect its expression in vitro.METHODS: The LEDGFp52 gene was cloned to adenoviral shuttle plasmid pAdTrack-CMV. Then, the resultant pAdTrack-CMV-LEDGFp52 was cotransfected into BJ5 183 bacteria with the adenoviral backbone plasmid pAdeasy-1. The adenoviral plasmid carrying LEDGFp52 was generated with homologous recombination in bacteria, and the adenoviruses were produced in 293 cells. These 293 cells were then infected with adenoviruses, and the expression of LEDGFp52 was detected by CPE (cytopathic effect) and western blot.RESULTS: The titer of Ad-LEDGFp52 adenoviruses was up to 5×1012 pfu/L after proliferation in 293 cells. LEDGFp52 was expressed efficiently in 293 cells after infection.CONCLUSION: The recombinant adenoviruses vector expressing LEDGFp52 was constructed successfully and can be used in further gene transfection experiments.

AIM: To construct and identify LEDGFp52 eukaryotic expression vector for RNA interference.METHODS: Recombinants were designed and established by targeting gene LEDGFp52 and plasmid pGensil-1 based on LEDGFp52 cDNA sequences of Genomes. Two pairs of oligonucleotides were synthesized according to the Tuschl principle and inserted into plasmid pGenSil-I to generate siRNA eukaryotic expression vector. DH5α strains were transformed, plasmids were extracted, and recombinant vectors were identified by the restriction map and the sequence analysis. The cultured cells were transfected by the recombinant plasmid (pGensil-1-RNA. LEDGFp52-1). At 48 hours after transfection, the whole cell protein was extracted, and the protein level was detected using Western blotting with mouse anti-human LEDGFp52 monoclonal antibody.RESULTS: Recombinant plasmids completely concord with the designs by the restriction map and the sequence analysis,the proteinlevel of LEDGFp52 was down regulated at 48hours after transfecting pGensil-1- LEDGFp52-1 expression vector into HeLa cells, the recombinant eukaryotic expression vectors were successfully constructed.CONCLUSION: siRNA recombinant can be successfully constructed by RNAi technique to inhibit the expression of LEDGFp52.

AIM: To construct the prokaryotic expression vector of rhLEDGFp52 gene,to obtain the rhLEDGF p52 protein.METHODS: rhLEDGFp52 gene was constructed into a prokaryotic expression vector pET30a (+) by recombinant DNA techniques and was identified by enzymatic digestion and sequence analysis. rhLEDGFp52 protein was induced expression by IPTG in E.coli BL21 (DE3), it was tested by Western blot and was purified by Ni-NTA His. Bind. Resin.RESULTS: We Successfully constructed the prokaryotic expression vector of rhLEDGFp52 gene and obtained its expression in E.coli BL21 (DE3),it was expressed in a soluble form and detected up to 34.63% of the total bacterial protein expressed in E.coli BL21 (DE3).Western blot analysis demonstrated that rhLEDGFp52 protein could spicifically integrate with LEDGF-ab. After purified by Ni-NTA His. Bind. Resin, the ultimate concentration of purified rhLEDGFp52 protein was 520μ g/ml and its purity was 87.93%.CONCLUSIONS: rhLEDGF p52 protein was obtained that provides an experimental basis for the further study of the biological function of rhLEDGFp52 protein.

Objective To study whether clinical variables could be used to predict the presence of esophageal varices(EV). Methods One hundred and twenty-six patients with decompensated liver cirrhosis were enrolled. Upper endoscopy was performed to identify the EV. The spleen vein (SV), portal vein(PV), spleen index(SI), ascites was determined by ultrasenography. Platelct count(Pt), prothrombin time(PT) and liver function was determined. Results Ninety-five patients with EV, and 42 patients with severe EV. Patients with EV had significant larger SI and lower Pt. Pt and SI were predictive factors for the presence of EV. When SI≥66.9 cm2 and Pt≤89.0×109/L, they had a positive predictive value of 97.4% and 96.5%, and a negative predictive value of 55.4% and 59.8%, respectively. SI was the only predictive factor for the presence of severe EV. When SI≥82.6 cm2, it had a positive and negative predictive value of 89.2% and 75.4%. Conclusions Pt and SI are predictive factors for the presence of EV. SI is the only predictive factor for the presence of severe EV. Non-invasive factors SI and Pt can be used to predict the EV in patients with decompensated liver cirrhosis.

Objective To study the advantage and disadvantage of two types of rat heart trans- plantation models.Methods Twenty Wistar rats served as donors and 20 SD rats as recipients ran- domly in two groups.Abdominal working and nonworking cardiac transplantation models from Wistar to SD rats were established.In the working model,the donors pulmonary artery was anastomosed to the left atrial appendage.The left ventricle ejected volume through an end to side anastomosis of the donor's aorta to the recipient's abdominal aorta.In the nonworking cardiac transplantation model,the donor's pulmonary artery was anastomosed to the recipient's inferior vena cava and the donor aorta was anastomosed to the recipient aorta.Results Working model and nonworking model were successfully established.The survival rate of both models was 90 %.Total operating time of the working model was 10 min shorter than the nonworking model.Postoperative UCG demonstrated the working trans- planted hearts had ejection.Conclusions Rat abdominal working cardiac transplantation model was used perfectly with high rate of survival and shorter total operating time.The working model approached cardiac physiological functions more than the nonworking model.

ObjectiveUsing large sample clinical study to reevaluate the safety and efficacy of recombinant human growth hormone(ANSOMONE) in treating pediatric growth hormone deficiency(GHD).Methods1012 cases were administered with ANSOMONE at a dose of 0.1 IU · kg-1 · d -1 in bedtime for 6 months.The alteration of mean height,growth rate,height standard deviation scores,bone age,and anti-GH antibody and occurrence of adverse effects were observed.ResultsThe growth rate was promoted from ( 2.50 ± 0.85 ) cm/year to ( 13.44 ± 4.27 ) cm/year,the net growth rate was( 10.94 ±3.69)cm/year.There was significant difference between the pre-treatment and post-treatment patients ( P ＜ 0.01 ).The post-treatment anti-GH antibody positive rate was 13.34％,which had no obvious effect on the efficacy.Few patients exert transient inflammation reactions,which disappeared with any treatment.ConclusionANSOMONE had significant efficacy,and could promote the height increase obviously.It had good safety without any obvious side effect and early-maturing effect.

Health Promotion ; Outcome and Process Assessment (Health Care) ; Workplace

Objective To explore the features of transsphenoidal microsurgical treatment of male pro- lactinoma and to evaluate the sexual function before and after transsphenoidal microsurgery.Methods A se- ries of 23 cases male prolactinoma were analysed combined with retrospective and prospective study.9 casese with sexual disfunction had finished the International Index of Erectile Function with 15-item questionnnaire (ⅡEF-15) before and after the surgery.Results 8/10 cases treated with dopaminergic agonist combined with surgery achieved complete remission,while only 4/13 cases achieved complete remission with surgery on- ly(x~2=5.490,P=0.036).Aecoding toⅡEF-15,sexual function improved after surgery in 9 cases(paired- sample) t test,P＜0.05).Conclusion Transsphenoidal microsurgical treatment of prolactinoma may im- prove the male sexual functions.Dopaminergic agonist combined transsphenoidal microsurgery may achieve a better outcome.

0.05).The combined use of sinomenine and CsA could prolong graft survival significantly(groups 4 vs groups 1,2,3,5)(P

Background Excimer laser corneal refractive surgery is widely used because it reduces the dependency of refractive error eye to glasses.However,the adverse effects of anti-inflammatory drugs is a problem after operation.Lotemax eyedrops is a kind of steroidal anti-inflammatory drug,with a good effect on the operative eye.But its influence on intraocular pressure(IOP) after long-term topical administration is less studied.Objective This study was to investigate the influence of lotemax eyedrops on IOP after long-term topical administration in myopic eyes received excimer laser corneal refractive surgery.Methods An one-month retrospective case serial study was adopted.The case history data of 1552 eyes from 1552 Patients who received excimer laser corneal refractive surgery were collected in Tangshan Eye Hospital from 2011 January to 2012 January,including 321 eyes with laser in situ keratomileusis(LASIK),608 eyes with sub-Bowman keratomileusis (SBK) and 623 eyes with femtosecond laser for flap excimer laser in situ keratomileusis(FS-LASIK).Lotemax eyedrops was topically administered after operation 4 times per day initial and then decreased 1 time weekly for 1 month.IOP was measured in before,1 week,2 weeks,3 weeks,4 weeks after operation with non-contact tonometer.The increasing range of IOP was divided into ＜5 mmHg,5-15 mmHg and ＞ 15 mmHg after operation.Optical head parameters were measured by Heidelberg retina tomography when IOP increasing after operation.Results Within 1 month,elevated IOP was found in 47 of 1552 eyes after administration of lotemax eyedrops,including 3.12％ (10/321) in the LASIK group,3.29％ (20/608) in the SBK group and 2.73％ (17/623) in the FS-LASIK group,without significant difference among them(x2 =1.95,P＞0.05).The IOP elevated 1 week after operation and remained normal after that till drug withdrawal in all eyes.There were no significant differences in the disk area (1.65 ±0.44) mm2 vs.(1.66 ±0.44) mm2,disk volume (0.40 ±0.09)mm3 vs.(0.39±0.08) mm3 and mean nerve fiber layer thickness (0.28 ± 0.08) mm vs.(0.29 ± 0.07) mm in IOP elevation duration compared with 1 month after IOP recovery(t =0.34,0.81,0.48,P＞0.05).Conclusions Topical administration of 0.5％ lotemax eyedrops after excimer laser corneal refractive surgery leads to reversible high IOP.It is very important to monitor the IOP during use of the drug.