Background Treatment of corneal ulceration by transplanting drug-loaded amniotic membrane has been used widespreadly abroad,however,seldom study is found in China up to now.Objective This study was to explore the sustained release property and the efficacy of the drug-loaded amniotic membrane.Methods The bacteriostatic area of amniotic membrane fragments immersed with different concentrations (5,20,30 g/L) of levofloxacin for different time points (5,15,30,60 minutes) was evaluated by in vitro test.Bacterial corneal ulceration models were established in 20 rabbits by injected 0.7 MCF staphylococcus aureus suspension (0.1 ml) into the central corneal stroma to form the cloudy area of 4.0-6.0 mm.Then the rabbits were randomized into two groups.Regular amniotic membrane transplantation was performed laterally and 0.5％ levofloxacin drops was topically administered after operation in the amnion+levofloxacin drops group,and drug-loading amnion transplantation was used in the drug-loading amnion group.Aqueous humor of 0.1 ml was collected in 30 minutes,1 hour,2,3.5,5.5 hours after levofloxacin was administered and 1 day,3,7,10,14,21 days after operation for the detect of levofloxacin level with high-performance liquid chromatography.The corneal symptom was scored based on McNeill's criteria in 1 week,2 weeks and 4 weeks and the ulceration area was assessed under the slit lamp in the first week.The pathological examination was carried out in the fourth week after surgery.Results The mean bacteriostatic area was bigger with the increase of levofloxacin concentration,and bacteriostatic area in amnion immersed for 15 minutes was bigger than that of 5 minutes (P＜0.01).The levofloxacin concentration of aqueous humor after transplantation was decreased by extending the time,and that in 30 minutes and 5.5 hours after operation was (0.873±0.264) mg/L and (0.106±0.027) mg/L,respectively,in the amnion+levofloxacin drops group,and that in day 1,3,7 after surgery in the drug-loading amnion group was higher than at 30 minutes in the amnion+levofloxacin drops group,showing all significant differences (all P =0.00).In the first,second and fourth week after operation,the corneal symptom score was 1.7±0.6,1.3±0.5,0.2±0.4 in the drug-loading amnion group and 2.2±0.8,2.0±0.6,1.5±0.8 in the amnion+ levofloxacin drops group,with the significant differences among the different groups and time points (Fgroup =9.49,P =0.01 ;Ftime =22.96,P=0.01).The corneal ulceration area was (1.6±0.6) mm2 in the drug-loading amnion group and (3.2±0.8) mm2 in the amnion+levofloxacin drops group 1 week after operation,showing a significant difference between them (t =3.98,P =0.00).Histopathological revealed that the various layers of cornea tissue appeared irregular arrangement in the amnion + levofloxacin drops group 4 weeks after operation with 1-2 layers of new squamous epithelium.Disorder hypothallus structure,more inflammatory cells and residual vascular cavity were visible.However,new squamous epithelium of 4-5 layers was seen in the drug-loading amnion group,and inflammatory cells and residual vascular cavity were less than the amnion+levofloxacin drops group 4 weeks after operation.Conclusions Levofloxacin-loaded amniotic membrane can sustained release levofloxacin and maintain an effective drug concentration in aqueous humor,which improves the treating efficacy for staphylococcus aureus corneal ulceration.

Six flavonol glycosides were isolated and calibrated from Ginkgo biloba extract, and then used to calibrate the content in 2 baiches of G. biloba reference extract, so was rutin. RSD values of rutin, kaempferol-3-O-rutinoside, kaempferol-3-O-rhamnoside-2-glu- coside, quercetin-3-O-rhamnop-yranosyl-2-O-(6-O-p-coumaroyl)-glucoside, kaempferol-3-O-rhamnopyranosyl-2-O-(6-O-p-coum-aroyl) - glucoside were around 1.1%-4.6%, nevertheless, RSD values of quercetin-3-O-glucoside and isorhamnetin-3-O-rutinoside were more than 5%. According to the results, the reference extract of G. biloba can be used as the substitute to determine rutin, kaempferol-3-O- rutinoside, kaempferol-3-O-rhamnoside-2-glucoside, quercetin-3-O-rhamnopyranosyl-2-O-(6-O-p-coumaroyl)-glucoside and kaempferol-3-0-rhamnopyranosyl-2-O-(6-O-p-coumaroyl)-glucoside instead of corresponding reference substances. So reference extract in place of single component reference in assay is feasible.
Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flavonols ; chemistry ; isolation & purification ; Ginkgo biloba ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo assess long term stent implantation efficacy of myocardial bridge and severe atherosclerosis lesions in the segments proximal to the myocardial bridge.

METHODSThe study population consisted of 3 groups (103 patients). Group A included 28 patients with severe atherosclerosis lesion of luminal narrowing of > or = 70% in the segments proximal to the myocardial bridge. Group B included 16 patients with symptomatic myocardial bridge lesion of systolic luminal narrowing of > or = 95%. Group C included 59 patients with severe atherosclerotic lesion of luminal narrowing of > or = 70%. All lesions were successfully treated with stent by standard interventional techniques. Quantitative coronary angiography was performed before and immediately after stent deployment. Follow-up Quantitative coronary angiography was performed at six months or later. Clinical evaluation was done at 20 months after PCI.

RESULTSThere was no significant difference in luminal diameter and stent diameter among 3 groups immediately after stent implantation (P > 0.05). At six months, restenosis occurred in 4 patients in Group A (14.3%), in 7 patients in Group B (43.7%), and in 8 patients in Group C (14.8%), respectively. The rate of restenosis was significantly lower in group A and C than in group B (P < 0.05). No significant difference was found between group A and C. Additional balloon dilating of stent were performed in all restenosis patients. Clinical evaluation at 20 months showed that all patients remained free of angina and cardiac events.

CONCLUSIONThe efficacy of intracoronary stent implantation in treating severe atherosclerosis lesion in the segments proximal to the myocardial bridge is not affected by abnormal haemodynamic changes of myocardial bridges. The rate of restenosis in intracoronary stent implantation of myocardial bridges is higher than that of atherosclerotic lesions in the segments proximal to myocardial bridge.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; Atherosclerosis ; therapy ; Coronary Artery Disease ; therapy ; Female ; Humans ; Male ; Middle Aged ; Myocardial Bridging ; therapy ; Stents ; Treatment Outcome

Diabetes is the most important comorbidity of cardiovascular disease, and cardiovascular disease is the main cause of mortality and disability of patients with type 2 diabetes. In order to standardize the diagnosis and treatment of patients with diabetes and cardiovascular disease, the National Health Commission Capacity Building and Continuing Education Center organized the experts from the field of cardiology and endocrinology systematically reviewing the research progresses and expert experiences of relevant disciplines from home and abroad, and formulated this consensus. This consensus covers the diagnosis, drug treatment, and risk factor management for patients with diabetes and cardiovascular disease (including atherosclerotic cardiovascular disease and heart failure) from the perspective of cardiovascular disease and diabetes management aiming to strengthen the comprehensive management of patients and ultimately to improve the prognosis of patients. The management of cardiovascular diseases mainly includes the management of blood pressure, blood lipids, anti-thrombosis, anti-myocardial ischemia, anti-ventricular remodeling and so on. Diabetes management mainly includes lifestyle intervention (including diet, exercise, weight loss, etc.), anti-hyperglycemia therapy (including drugs and insulin), blood glucose monitoring, and hypoglycemic prevention. In addition, specific clinical recommendations are given to patients with special health care needs such as diabetic nephropathy, elderly (>75 years), and cardiovascular critical illness.

Objective To explore the indications of axillary artery approach in endovascular treatment and to analyze complications associated with axillary artery puncture.Methods In 111 cases endovaccular treatment via axillary artery approach by Seldinger technique was performed.The indications of axillary artery approach and the complications associated with axillary artery punture were respectively analyzed.Results The success rate of angiography was 100％,and the success rate of angioplasty was 90.8％ by axillary artery approach.The total incidence of complications was 10.3％.The incidence of local hematomas was 4.8％,nerve injury was 3.2％,pseudoaneurysm was 0.8％,acute thrombosis of the axillary artery was 0.8％,acute thrombosis of the axillary vein was 0.8％.The main factors affecting complications include vascular conditions,perioperative medication,anatomy of the axillary artery,location of puncture point,the success rate of first attempt,and pressure of bandage.Conclusions The axillary artery approach increases the success rate of endovascular treatment.Reasonable choice of axillary artery appruch,meticulous perioperative management and fully understanding the anatomical characteristics of the axillary artery can decrease the complications of axillary artery puncture.

Objective To discuss the methods, advantages and indications of multipoint puncturing in performing endovascular therapy for complex lower extremity arterial occlusive diseases. Methods During the period from Oct. 2011 to Oct. 2013, a total of 46 patients with complex lower extremity arterial occlusive diseases were treated with endovascular therapy by using multipoint puncturing technique. The puncturing type, the advantages of multipoint puncturing technique and the success rate of this technique were analyzed. Results The multipoint puncturing was divided into three types: type Ⅰ : puncturing from the opposite direction to deal with the same target vessel; type Ⅱ: direct puncturing of the target vessel; and type Ⅲ:using the same puncturing direction to deal with different target vessels. The success rate of endovascular treatment was 80.4%. Conclusion Multipoint puncturing technique helps improve endovascular treatment success rate for lower extremity arterial occlusive diseases. Full understanding of the advantages of multipoint puncturing technique, perfect preoperative planning, precise puncturing technique and proper interventional equipments are helpful to ensure a successful treatment.

Objective To investigate the effects of distal thrombolysis versus thrombus aspiration on myocardial perf usion and prognosis in patients with acute ST segment elevation myocardial infarction(AMI)during emergency percutaneous coronary intervention(PCI). Methods 96 patients with acute ST segment elevation myocardial infarction(onset<6 hours)were randomly divided into thrombolysis group and distal thrombus aspiration group. Patients in the distal thrombolysis group(n=46)received transcatheter urokinase injection to the occlusive segment followed by balloon dilatation or stenting. Patients in the thrombus aspiration group(n=50)were given thrombus aspiration after balloon angioplasty or stenting. Patients were followed up for 30 days after operation. The coronary blood flow and myocardial perfusion were compared between the 2 groups. The incidence of major adverse cardiac events(MACE)and left ventricular systolic function after 30 days were compared. Results There was 1 case in the distal thrombolysis group (0.2%)and 6 case in the thrombus aspiration group(12%)presented with TIMI fl ow≤grade Ⅱ(P=0.008). A 65.2% of patients in the distal thrombolysis and 42.0% of patients in the thrombus aspiration group achieved > 50% of ST segment resolution in 2 hours(P=0.019). At 30-day follow up,the LVEF was found higher in the thrombolysis group compared with the aspiration group(54.1±8.6)% vs.(50.8±7.3)%,P=0.047 but the LVEDD(44.3±7.2)mm vs.(46.5±6.8)mm,P=0.038 and NT-proBNP levels(117.8±71.8)μg/L vs.(161.2±72.3)μg/L,P=0.025 were found significantly lower in the thrombolysis group. Conclusions For the ST segment elevation myocardial infarction,distal thrombolysis,when compared with thrombus aspiration,may reduce the incidence of slow flow and no reflow,and may improve the left ventricular systolic function.

Objective To discuss the key points of endovascular therapy for complex subclavian artery occlusive diseases. Methods During the period from January 2012 to December 2013, a total of 92 patients with complex subclavian artery occlusive disease were admitted to Xuanwu Hospital of Capital Medical University, Beijing, China. The clinical data were retrospectively analyzed. The features of the lesions, the success rate of endovascular therapy, the use of combined approaches, the relief of symptoms after treatment, etc. were evaluated. Results The complex subclavian artery occlusive diseases could be divided into three types. Type Ⅰ: long segment of the left subclavian artery was occluded; type Ⅱ: ostial stenosis or occlusion of the right subclavian artery; and type Ⅲ: subclavian artery stenosis or occlusion was associated with the ostial disorder of the vertebral artery, or the opening of vertebral artery was affected by the subclavian artery stenosis or occlusion. The technical success rate was 82.6%. Combination use of femoral artery and brachial artery approach was employed in 27.2% of patients, which had improved the technical success rate. After the treatment the symptom improvement rate was 81.6%. Conclusion Upper limb artery approach can improve the re-canalization rate of left subclavian artery with long segment occlusion, and can ensure the accurate positioning of stent at the site of right subclavian artery opening. During the procedure of endovascular intervention for subclavian artery occlusion disease, attention should be paid to the protection of the vertebral artery.

Brain ; N-Methylaspartate ; Neurons ; Patch-Clamp Techniques

OBJECTIVETo establish normally conditionally-immortalized human umbilical vein endothelial cells (HUVECs) by ectopic expression of the human telomerase catalytic enzyme (hTERT) and simian virus 40 large T (SV40 LT) antigen.

METHODSPrimary HUVECs were transfected with recombinant retrovirus containing hTERT or SV40 LT respectively. Subsequently drug resistant cell clones were screened and expanded for further studies. Endothelial cell biomarkers were confirmed by examination.

RESULTSThe morphological phenotype of the transfected cells was similar to the non-transfected cells. Von Willebrand factor, hTERT and SV40 LT could be detected in transfected HUVECs. Moreover, higher telomerase activity in transfected cells was maintained for over 50 population doublings compared with only low level of endogenous telomerase transiently at early population doublings in primary HUVECs. When exposed to TNF-alpha (tumor necrosis factor-alpha), the expression of E-selectin in transfected cells was significantly up-regulated, but no alteration of endothelial lipase was found.

CONCLUSIONEctopic coexpression of hTERT and SV40 LT can effectively immortalize HUVECs without tumorigenicity in vitro. Immortalized HUVECs may be an ideal target of further molecular function studies.

Antigens, Polyomavirus Transforming ; genetics ; metabolism ; Cell Culture Techniques ; methods ; Cell Size ; Cell Survival ; physiology ; Cells, Cultured ; DNA-Binding Proteins ; genetics ; metabolism ; Endothelial Cells ; cytology ; physiology ; Genetic Enhancement ; methods ; Humans ; Protein Engineering ; methods ; Recombinant Proteins ; metabolism ; Telomerase ; genetics ; metabolism ; Tissue Engineering ; methods ; Transfection ; methods ; Umbilical Veins ; cytology ; physiology