BACKGROUND:Although the methods of mechanical scrapping,tissue block transplantation,and enzyme digestion to culture endothelial cells have been quite mature,it has been constantly updated.OBJECTIVE:To explore the culture and identification of rabbit aortic endothelial cells.METHODS:Aorta was derived from one-week-old New Zealand rabbit.Following removing the adventitia,the endothelium was digested in the mixture of 2 g/L type Ⅰ collagen,2 9/L type Ⅱ collagen,2 g/L type Ⅳ collagen,and 2 g/L type Ⅴ collagen (1:1:1:1)for 20 minutes,and the digestionwas stopped with culturemedium(1:1).Cells in the endothelium were lightly scraped to make cell suspension,which was centrifuged to precipitate cells with DMEM culture medium containing 20%fetal bovine serum,1 μg/L VEGF,2 μg/L bFGF,and 6 U/L gentamicin.The cells were then blown into single cells and the culture liquid was changed every 48 hours.The cells were passaged according to the ratio of 1:2.Inverted phase contrast microscope was used to observe cell culture;immunohistochemistry and immunofluorescence were used to determine Ⅷ-related antigen;electron microscope was used to detect Weibel-Paladed body.RESULTS AND CONCLUSION:Ⅷ-related antigen and electron microscopy confirmed that the primary and 5-passage endothelial cells were successfully cultured,suggesting that rabbit aortic endothelial cells were cultured into cell line.Ⅷ-related antigen and electron microscopy determined that a combination with W-P body was a good method to determine endothelial cells.

Objective To explore the effectiveness and safety of fiberbronchoscope combined with cryotherapeatic equipment in treatment of bronchial foreign body in children.Methods 72 children with bronchial foreign body received fiberbronchoscope combined with cryotherapeatic equipment, then the complications were observed during and after the operation.Results Bronchial foreign bodies were successfully removed in all the patients. Among the 72 cases, transient hypoxia was the most common complication during the operation which happened in 37 cases and most occurred in the small age group, and decreased with the age increasing; cough or worsening of cough was the most common complication after the operation which happened in 20 cases; No serious complications were found during the follow-up.Conclusion Although it has some complications when bronchial foreign body received the fiberbronchoscope combined with cryotherapeatic equipment, but the operation time is short, and the complication is mild. It has great application value and security.

Objective To establish a rapid specific quantitative assay for Bacillus anthracis detection. Methods According to the principle of complex probe quantitative assay, the primers and quantitative probes targeted at chromatosome DNA rpoB were designed and applied to detect Bacillus anthracis. The influence factor of quantitative PCR were determined. Results The optimal system of this method was aquired: the length of quenching probe is 15mer,the ratio of fluorescent probe to quenching probe is 1/2 and the concentrtion of Mg 2+ is 3 mmol/L.The sensitivity of this assay for Bacillus anthracis is 10 3 copies. It can distinguish Bacillus anthracis from other closely related Bacillus. Conclusion The method can rapidly quantitatively detect the Bacillus anthracis with high sensitivity and specificity, it can be applied to clinical diagnosis.

Objective To investigate the manifestation and the prevention means of video display terminal (VDT) visual fatigue in old people. Methods Conventional eye inspection were performed for 45 patients, including optical muscle movement, position of eye, confluence function and presbyopia. Results We found that the average difference between middle distance correcting lens and 33 cm reading distance from presbyopia correcting lens was about 0.52 DS( P

Objective To evaluate the expression of molecular markers in prostate cancer and to clarify the significance of these markers as prognostic indicators for androgen deprivation therapy. Methods A series of 116 prostate cancer patients under androgen deprivation therapy as a single treatment was reviewed. Expression levels of 7 proteins, including androgen receptor(AR),E-cadherin, Chromogranin A(CgA) , Ki-67, Survivin, EZH2 and hepsin, were measured by immunohistochemical staining. Results Of the 7 molecules. Ki67,EZH2 and Survivin expression were significantly associated with several conventional prognostic factors. Univariate analysis identified clinical stage, Glea-son scores,pretreatment serum PSA level, Ki-67 and Survivin expression as significant predictors for prostate-specific antigen (PSA) progression after endocrine therapy. Of these significant factors, Survivin expression, clinical stage and Gleason scores appeared to be independently related to PSA progression after endocrine therapy by multivariate analysis. Furthermore, there were significant differences in PSA progression-free survival according to positive numbers of these three independent risk factors. Conclusion Survivin could be a useful independent prognostic factor in prostate cancer with endocrine therapy, besides clinical stage and Gleason score.

Objective To discuss the relationship between serum anti-Phosphalipase A2 receptor (PLA2R) antibodies and glomerular IgG4 subclass in patients with membranous nephropathy and evaluate the diagnostic value of the two markers.Methods Patients diagnosed as membranous nephropathy from October 2011 to April 2014 in Peking Union Medical College Hospital were included and divided into IMN and SMN groups accoding to their clinical diagnosis.Serum anti-PLA2R antibodies and glomerular IgG subclasses were both detected by indirect immunofluorescence assay.Receiver operator characteristic curves were used to evaluate the diagnostic efficiency of anti-PLA2R antibodies and glomerular IgG4.Results Prevalence of serum anti-PLA2R antibodies of IMN patients was 69.5％ (41/59); prevalence of MLN patients was 4.8％ (1/21).Within the IMN group,thirty-five patients showed positive results of both serum anti-PLA2R antibodies and glomerular IgG4; Six patients were positive for serum anti-PLA2R antibodies but negative for glomerular IgG4; Seventeen patients were positive for glomerular IgG4 but negative for serum anti-PLA2R antibodies; one patient was negative for both tests.The sensitivity of serum anti-PLA2R antibody was 69.5％ and the specificity was 95.2％; the sensitivity of glomerular IgG4 was 89.8％ and the specificity was 52.3％.The sensitivity of the combined marker consisting of serum anti-PLA2R antibody and glomerular IgG4 was 59.3％ and the specificity was 100％.Four out of the six patients secondary to HBV infection,one out of the three patients secondary to Sj(o)gren syndrome,one out of the three patients secondary to malignant tumor showed positive results of serum anti-PLA2R antibodies.Conclusions Serum antiPLA2R antibodies were of high prevalence among IMN patients; the prevalence among SMN patients varied with etiologies.Results of serum anti-PLA2R antibodies and glomerular IgG4 were helpful to rule out secondary etiologies in the diagnosis of membrnous nephropathy.

Objective To evaluate the efficacy and safety of small-middle endoscopic sphincterotomy combined with endoscopic papillary balloon dilation for patients with extrahepatic bile duct stone. Methods The patients with special duodenal papilla included 38 cases, and those with normal duodenal papilla 143 cases. 38 patients had received SEST + EPBD, 143 had received MEST + EPBD. Results 181 patients had received SMEST + EPBD in our hospital and the related data were retrospectively analyzed. All 181 patients were successfully removed, the success rate was 100.00 %. There was no perforation occurred postoperatively, but mild acute pancreatitis occurred in 8 patients (4.42 %, 8/181) and bleeding occurred in 9 patients. The three major early complications rate were 9.93 %(17/181), which was cured by the conservative management. Conclusion SMEST plus EPBD is a safe and effective treatment for extrahepatic bile duct stone, with retaining the feature of sphincter of duodenal papilla, especially for patients with special duodenal papilla.

Objective To investigate the traffic capacity of ambulances in Beijing,and to explore factors and methods to resolve this problem.Method A survey on all the ambulances on-duty in Beijing from 17th August to 17th September 2006 was conducted by questionnaires.Results The average speed of ambulances in Beijing was 32.07 km/h.There were statistically significant differences in terms of the areas and time, respectively.Conclusions The traffic jam of Beijing was serious.The traffic capacity of ambulances was far from being ideal,which restricted the operation of 120 ambulances.It is difficult to satisfy the needs of Beijing 2008 Olympic games and it is time for the relevant authorities to search for appropriate methods and solve this problem.

After testing the resistance of streptomycin to NS-41-80 Streptomyces Nanchangensis NS-41-80, the spores treated with 4 different dosage of super-mutagent EMS were regenerated on the lethal media, thus 3,000 streptomycin-resistant ( str ) str mutants were obtained. Then 202 spore mutants were screened through the pre-screening experiments and their yields are beyond 20% higher than the original strains respectively. In the re-screening experiments 48, 7 and 1 mutants were obtained whose yields are 100%, 200% and 300% higher than the original strains respectively and take the precentage of 23.76% and 1.60%, 3.46% and 0.23% and 0.5% and 0.03% of the number of the pre-screened strains and the rescreened strains respectively. Then 5 mutants were fermented whose yields are above 240% higher and the original strain through the continuous 3 batches fermentation, the yields are 57%～96.4% more than the original strains. Among them the yield of 80-5.3-165 is upwards of 6,000?g/mL and its norm is 5,855?g/mL. And a model of the streptomycin-resistance mutational screening of the strain producing Nanchangmycin was established.

Objective To explore the clinical outcome of improved technique of Gamma nail in the treatment of intertrochanteric hip fracture of the elderly patients. Methods From March 2002 to October 2006.39 patients with intertrochanteric hip fracture were operated by improved technique of Gam-ma nail.There were 18 males and 21 females at average age of75.7 years(67_98 years).There were 6 patients with type A1 fracture,24 with type A2 fracture and 9 with type A3 fracture according to AO/ASIF classification.Of all.36 patients(92.3%)had osteoporosis.The operation improvements included the following points:(1)The patients were placed at the lateral decubitus position with the fractured limb on the uppermost,with flexion of knee and hip of 60°.The normal hip and knee were flexed as possible.(2)One-off indirect traction reduction was used after general anesthesia. no requirement of continuous mechanical traction.(3)C-arm image intensifier was employed to obtain normal and lateral projections.Results Of all,35 patients were followed up for a mean period of3 years and 2 months, ranging from 6 months to 5 years and 2 months.Operation data showed incision length of(4.3±1.2)cm,mean opera-tion time of(46±10)minutes,intraoperative bleeding volume of(65±26)ml and intraoperative X-ray exposure of(3.0±2.1)times.Postoperative recovery data showed survival in one-year follow up,with ambulation time of(10.5±3.6)days and fracture union time of(10.9±2.1)weeks.Mean Parker's score wag(6.9±3.2)points 6 months after operation. Conclusions Improved technique of Gamma nail can shorten operation duration,reduce operative trauma and bleeding,reduce X-ray exposure and im-prove success rate of surgery.as facilitates early pest-operative recovery and reduces the perioperative mortality rate of the elderly.