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 investigate the anti-tumor effects of combined recombinant murine interleukin-12 and GM-CSF gene adenovirus vector(AdCMVIL-12,AdCMVGMCSF) on B16 melanoma.Methods B16 melanoma cells were injected subcutaneous at right back of C57/BL6 mice,48 tumor bearing mice were divided into 4 groups at random,when tumor diameter attained to 5 mm,AdCMVIL-12,AdCMVGM-CSF and AdCMVIL-12+AdCMVGM-CSF and AdCMVLacZ(control) were injected in different groups,tumor growth and immune responses of each group were tested,and the IL-12 and GMCSF levels in serum were observed.Results In combined group,necrosis, lymphocytes and macrophages were found in tumor tissues.30 d after treatment,the average volume in combined group was(60.73?11.36)mm~3, compared with AdCMVLacZ(control) group,and AdCMVIL-12 group,AdCMVGM-CSF group [(675.18?80.59),(186.91?19.15),(223.45?23.11)mm~3],there were significant differences(P

Objective To investigate the clinical application of specific allergen immunotherapy (AIT)for the children with al-lergic asthma and rhinitis.Methods The in vitro specific allergen test system was adopted to detect allergen specific IgE and total IgE in children outpatients of the Children's Hospital.The children patients with dust mite allergy were divided into groups for con-ducting therapy.The immunotherapy group adopted the Dermatophagoides farinae drops for hypoglossis desensitization therapy, while the control group was performed the anti-allergic therapy by the common drugs.The clinical control indicator scores were per-formed after 6-months and 12-month therapy and at the same time the change of serum specific LgE was observed.Results Among 218 cases of allergy,114 cases(52.4%)were inhalation allergy and 101 cases (46.3%)were food allergy.98 cases of dust mite al-lergy accounted for 44.95% of the total allergy and 85.96% of the inhalation allergy.The difference in the curative effects between the AIT group and the control group had statistical significance(P <0.01).Conclusion Among 218 children cases of allergy,the dust mite allergy was maximum in the specific allergen detection;the hypoglossis specific immunotherapy has significant curative effect for the children with allergic asthma and rhinitis.

Of 807 Cases of Diabetes mellitus, 100 were ketoacidosis with or with-ot coma(40 Cases among them with coma), 55 were males, 45 females. Therange of age was 5 to 76 years. There were 39 cases of ketonemia with ju-venile diabetes, and the incidence was 29. 5 per cent of 137 cases. 61 werefound to have ketonemia with adult dialetes, the incidence being 10. 2 percent of 594 cases. Blood glucose level was 166. 7 to 1000mg/dl. The mostcommon precipitating causcs of ketonemia were infection and the dis-continue of insulin treatment(58 per cent). According to the dosage of insu-lin, our patients were divided into two groups. One was a small dosagegroup(the total mean dose 25. 5 ? 3. 1 u), the other was a large dosage group(the first 8 hours' mean dose 121. 67 ? 102u, the fitst 24 hours' mean dose206. 62 ? 18. 01) There were on death and severe complications of insulintherapy in the small dosage group, but 13 (16 per cent) were died in thelarge dosage group, in which there took place such complications as hypo-kele mia(11 per cent), hypoglycemia (26 per cent) and encephaledema (3. 8per cent). Also, the negative acetone bodies occured faster in the smalldosage group. For this reason, we consider the effect of the treatment inthe small dosage group was better. Recently, we have controlled alkelidosage more strictly than before. 100-200ml of 4% Bicarbonate was givento the patient if the blood CO_2 combinedpower was lower than 20 Vol pe,

Objective To study the etiology and relative factors of dilated cardiomyopathy (DCM). Methods A total of 233 DCM patients were enrolled and conducted epidemiological survey. Stable angina patients in the same region were collected as control group. The epidemiological investigation included: life styles and history of diseases; blood pressure, electrocardiogram and echocardiography; serum lipids, glucose, hs-CRP, cTNI and NT-proBNP. Results The prevalence rate of DCM was 29.1 per 100 000 population, the male being almost 2-fold more involved than female, and farmers accounted for 77.7%. Familial clustering was not obvious. There were low level of history of hypertension (12.4%), ischemic heart disease (2.1%), myocarditis (5.2%) and diabetes (3.4%). Compared with control group, DCM patients had higher proportion of farmer (60.0% vs 77.7%, P

Objective To investigate the effect of different treatment on ST-elevation myocardial infarction (STEMI) by evaluating the changes of myocardial collagen metabolism. Methods Forty-one patients with STEMI were divided into drug treatment group (22 cases) and selective percutaneous coronary intervention (PCI) group (19 cases). Meanwhile,47 healthy controls were included in control group. The levels of serum carboxy terminal propeptide of type Ⅰ procollagen (PⅠCP), precollagen Ⅲ ( PC Ⅲ ), matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) were measured by enzyme-linked immunosorbent assay at the 3rd ,6th, 12th, 18th month after STEMI,and compared with those in control group. Results The level of P Ⅰ CP in selective PCI group was significantly lower than that in drug treatment group at the 3rd, 6th month [( 15.08 ± 3.37 ) μ g/L vs. ( 19.78 ± 2.22 ) μ g/L, (23.88 ± 3.33 ) μg/Lvs. ( 30.00 ± 3.14) μ g/L, P ＜ 0.05], but there was no significant difference at the 12th and 18th month (P ＞0.05 ). The level of serum PC Ⅲ in selective PCI group was significantly lower than that in drug treatment group at the 3rd month [(50.70 ±4.83) ng/L vs. (59.91 ±4.64) ng/L,P ＜0.05],and there was no significant difference at others times between drug treatment group and selective PCI group. There was no significant difference in P Ⅰ CP/PC Ⅲ between drug treatment group and selective PCI group in all times(P ＞0.05). Compared with those in control group,the levels of serum MMP-1 in all times were significantly decreased in drug treatment group and selective PCI group(P ＜ 0.05 ), but there was no significant difference between drug treatment group and selective PCI group at same times (P ＞ 0.05). The level of serum TIMP-1was significantly higher in selective PCI group than that in drug treatment group at the 3rd and 6th month [(61.89 ± 11.44) μg/L vs. (52.23 ±4.97)μg/L, (62.85 ±6.31) μg/L vs. (52.97 ±6.43)μg/L,P＜0.05] ,and there was no significant difference in TIMP-1 between drug treatment group and selective PCI group in other times (P＞ 0.05). MMP-1/TIMP-1 in selective PCI group was significantly lower than that in drug treatment group at the 6th month [( 1.53 ± 0.25 )% vs. ( 1.95 ± 0.04)% ,P ＜ 0.05] ,and there was no significant difference in MMP-1/TIMP-1 between drug treatment group and selective PCI group in other times (P ＞0.05). Conclusions Selective PCI can improve myocardial collagen metabolism in patients with STEMI in short term ( ＜ 6 months), but it doesn't appear to be superior to drug treatment in long term (6 - 18 months). Patients with STEMI should enforce drug treatment if they have not been treated with PCI.

The high frequency subject terms were extracted from the PubMed-covered papers published from January 2000 to March 2015 on text mining of biomedical field to generate the matrix of high frequency subject terms and their source papers.The co-occurrence of high frequency subject terms in a same paper was analyzed by clustering analysis.The hotspots in text mining of biomedical field were analyzed according to the clustering analysis of high frequency subject terms and their corresponding class labels, which showed that the hotspots in text mining of bio-medical field were the basic technologies of text mining, application of text mining in biomedical informatics and in extraction of drugs-related facts.

Objective To evaluate the efficiency and safety of domestic rapamycin-eluting stent(Firebire stent) in the treatment of coronary heart disease.Methods Coronary angiography and stenting were carried out through femoral artery approach in 331 patients with coronary heart disease.The incidences of in-hospital major adverse cardiac events were recorded all the patients were followed up for 1 year.The follow-up angiography was carried out at at 6-9 months after discharge.Results A total of 597 domestic rapamycine-eluting stents were implanted in 331 patients.One patient died of pump failure after primary PCI and another patient required emergency CABG because of coronary artery rupture.NSTEMI happened in 3 patients after PCI.The incidence of in-hospital MACE was 1.5%.Three hundred and one patients finished a mean follow-up of 12.3?3.2 months.During the follow up period,non-fatal myocardial infrarction was recorded in 1 case,recurrent angina in 16 cases and target lesion revascularization in 5 cases.The overall MACE rate was 4.3%.Follow-up angiography was completed in 110 patients and the restenosis rate was 5.5%(6/110).Conclusion Domestic rapamycin-eluting stent(Firebird stent) was safe and effective in the treatment of coronary heart disease and may provide a higher cost-effectiveness due to its relatively low price.However,its long-term effect needs more studies to verify.

Objective To investigate the relationship between cerebral 0, metabolism during coronary artery bypass graft (CABG) performed with cardiopulmonary bypass (CPB) and postoperative mental disorders (POMD) in the elderly.Methods Thirty patients (13 male,17 female) aged 65-76 yr undergoing CABG performed with CPB were studied. Patients with senile dementia were excluded. The patients were premedicated with scopolamine 0.3 mg and pethidine 50mg. Anesthesia was induced with midazolam 0.05-0.1 mg?kg_(-1), propofol 0.2-0.5 mg?g_(-1), fentanyl 10-15?g?g_(-1) and vecuronium 0.1-0.3 mg?g_(-1) and maintained with isoflurane inhalation and intermittent iv boluses of fentanyl. Nasopharyngeal temperature (T) was maintained at 26℃-28℃) during CPB. Radial artery was cannulated for BP monitoring and blood sampling.Swan-Ganz was placed via right internal jugular vein (UV).Another catheter was inserted into right UV and threaded retrogradely until J point for blood sampling. Blood samples were taken from artery and UV simultaneously for blood gas analyses and determination of blood glucose and lactate concentration before anesthesia (T_1 ),immediately after induction of anesthesia (T_2), during splitting of sternum (T_3) when T was reduced to 28℃ (T_4) and rewarmed to 35℃(T_5 ) and at the end of bypass (T_6 ).Cerebral O2 delivery (CDO2 ),cerebral oxygen consumption (CMRO2),cerebral glucose consumption (CMRglu) and lactic acid production (ADVL) were calculated. POMD was assessed using CAM-ICU chart.Results (1)Seven patients developed POMD (7/30 or 23.3% ).(2) Low jugular bulb 02 saturation (SjO2 ≤50% or PjO2≤25% mm Hg) was observed in 8 patients during rewarming. The incidence was significantly higher in POMD patients (3/7 or 42.96%) than that in non-POMD patients (5/23 or 21.74%). (3) CBF/CMRO2 was significantly lower in POMD patients than in non-POMD patients during rewarming.(4) CMRO2/CMRglu was significantly lower but ADVL was significantly higher in POMD patients than in non-POMD patients during hypothermia, rewarming and at the end of CPB.(P

The aim of this study was to evaluate the accuracy of transtracheal doppler (TTD) cardiac output monitoring. Eleven mongrel dogs (weight: 8～15kg) were anesthetized with intravenous thiopental 20mg?kg~(-1), and the doppler tube was inserted into trachea and fixed at a proper position where clear signal was received, and the catheter with thermistor was passed into pulmonary artery via femoral vein. CO were measured simultanously by thermodilution (TD) and TTD in the basic state, during 10 mins after intravenous administration of propofol 2mg?kg~(-1), in 30 mins following intravenous dopamine 100?g?kg~(-1) or under condition of isovolumic hemodilution in sequence. The results demonstrated that the CO measured by TTD correlated positively with that by TD (r=0.88, P