Objective:The aim is to establish L-glutamate specific aminotransferase-L-glutamate dehydrogenase coupling 96-well high throughput screening method,which is applied to molecular evolution of aminotransferase WecE from E.coli.Methods:An optical assay for aminotransferase catalytic activity based on aminotransferase-glutamate dehydrogenase coupling system is established by optimization of coupling enzyme loading,signal molecule NADH concentration and coupling time.Mutants library of WecE is obtained by sitedirected saturation mutagenesis.Positive mutants can be screened out through 96-well preliminary screening and flask second screening.Results:The target transamination reaction is coupled with L-glutamate dehydrogenase indicative reaction system which consists of 0.5 U/ml enzyme loading and 0.4 mmol/L NADH.A positive mutant Y321F whose catalytic activity increases 3.4 fold compared to that of wild type is screened out in Tyr 321 saturation mutagenesis library of WecE.Conclusion:An accurate high throughput screening method with weak background interference is established.It offers feasible solution for molecular evolution of L-glutamate specific aminotransferase.

Objective To compare the effect after endovascular repair (EVAR) or open repair (OR) of ruptured abdominal aortic aneurysm (rAAA) in Department of Vascular Surgery,Beijing Anzhen Hospital.Methods Clinical data of 46 repaired rAAAs patients was retrospectively analyzed from 2005 to 2015.The difference between the EVAR group and the OR group in perioperative mortality,operation time,ICU stay,blood transfused,length of stay (LOS),complication rate were compared by x2 test and t test.Results 18 rAAA patients were repaired by EVAR,aged from 51 to 91 with a mean of (68 ±9).28 were repaired by OR,aged from 41 to 83 with a mean of (70 ± 11).Perioperative mortality was 21.0％ for EVAR and 28.6％ for OR (P ＞0.05).LOS was (15.3 ±9.5) days for EVAR,and (23.9 ± 10.5) days for OR (P ＜0.05).Blood transfused was (3 210 ± 3 780) ml for EVAR and (4 814 ± 3 392) ml for OR (P＜0.05).ICU stay time was (7.7 ±4.2) d for EVAR and (4.2 ±2.5) d for OR (P＜0.05).Conclusion EVAR is a reliable approach for the treatment of acute rAAA.

Objective To evaluate endovascular therapy and open surgery for subclavian artery occlusion disease. Methods In this study, 69 patients received endovascular therapy (44 patients)or open surgery(25 patients)from January 2002 to July 2007.Balloon dilatation was carried out in 3 cases and 43 stents was placed in 41 cases. Results All procedures were successful. In endovascular therapy group, the ratio of healthy/dieased side of mean blood pressure was improved from 0.66±0.14 to 0.96±0.13(t=9.532,P＜0.001＝;in surgery group, the ratio improved from 0.63±0.16 to 0.95±0.18(t=8.236,P＜0.001＝.Sixty-one discharged patients were followed up for 2～49 months(mean 16.7 months),in endovascular group, restenosis occurred in 1 patient 1 year after the therapy, in surgery group, all prothesis remained patent and there was no complication related to prothesis. Conclusions Both endovascular therapy and surgery were the effective methods for subclavian artery occlusion, and endovascular therapy is preferred for less invasiveness.

Objective To investigate the treatment and prevention for postoperative complications of abdominal aortic aneurysm(AAA)in open surgery.Methods 329 AAA patients received open surgery from January 1991 to August 2009.The postoperative complications were analyzed retrospectively.Results 30 d mortality rate was 0.91%,the incidence of postoperative complications was 19.1%(63/329),including cardiac dysfunction in 21 cases,respiratory insufficiency in 15 cases,myocardial infarction in 6 cases,renal failure in 5 cases,arrhythmia in 6 cases,cerebral infarction in 2 cases,artery embolism of lower extremity in 2 cases,wound dehiscence in 2 cases,incisional hernia in 1 case,ecchymoma in 1 case and deep vein thrombosis in 2 cases.One patient died of acute myocardial infarction,one died of renal failure after 20 d dialysis,1 patient died of premature ventricualr contraction and fibrillation ventricular.Other patients recovered well. Conclusions Cardiac dysfunction and respiratory insufficiency are the main postoperative complications of AAA.Preoperative evaluation.careful intraoperative maneuvre and postoperative care ale the key to improve the treatment effectiveness.

Objective To evaluate surgical therapy for ruptured aortoiliac artery. Method Between Apr 1984 and Dec 2003, 23 patients of ruptured aortoiliac artery were admitted with ruptured aortoiliac artery aneurysm in 18 cases and traumatic artery rupture in 5 patients. Two patients were treated with direct vascular repair and 21 patients underwent prosthetic grafts replacement. Result Four cases died perioperatively including 2 patients dying of acute renal failure, one of upper gastrointestinal heamorrhage caused by stress ulcer at 48 h after operation, and one of respiratory failure in 72 h. Conclusion Based on the etiology emergency operations should be performed on patients of ruptured aortoiliac artery by repairing or prosthetic grafts replacement.

Objective:To compare the perioperative complications of carotid endarterectomy with patch angioplasty or primary closure.Methods:The clinical data of 492 carotid endarterectomy patients at the Vascular Surgery Department of Anzhen Hospital from Mar 2003 to Dec 2016 was analyzed retrospectively.Results:There were 364 cases (74%) in the patch angioplasty group and 128 cases (26%) in the primary closure group. The incidence of perioperative ischemic stroke was significantly lower in the patch angioplasty group than that in the primary closure group (0.8% vs. 3.9%, P=0.031), and there was no difference in the incidence of the remaining perioperative complications. By subgroup analysis, the incidence of perioperative ischemic stroke was significantly lower in the patch angioplasty group than in the primary closure group when the diameter of the internal carotid artery was <5 mm (0.7% vs. 6.0%, P=0.001), whereas there was no difference between the two groups when the diameter of the internal carotid artery was ≥5 mm. Conclusions:Carotid endarterectomy with patch angioplasty can reduce the incidence of perioperative cerebral infarction, especially in cases with an internal carotid artery diameter <5 mm.

Objective To evaluate retrograde transpopliteal access for femoral-popliteal artery total occlusion with blind puncture.Methods Clinical data of 22 cases admitted from Sep 2014 to June 2016 undergoing endovascular treatment of the femoral-politeal artery occlusion with transpopliteal artery retrograde access by blind puncture were analyzed.Results A total of 22 patients underwent retrograde popliteal access with blind puncture after antegrade attempts failed.Puncture above the knee was performed in 18 cases and below the knee in 4 cases.The average increase of ABI was 0.57.Hematoma of puncture site was observed in 2 patients,other complications included pneumonia in 1 case and renal insufficiency in 2 cases.The mean follow-up time was (13 ± 5)months.Restenosis occurred in 8 patiens(36.4％)during the follow-up time.The primary patency was (86.4 ± 0.07) ％ at 6 months and (70.7-± 0.12) ％ at 12 months.There was no major amputation rate and mortality during the follow-up.Conclusions Retrograde transpopliteal access for femoral-popliteal artery occlusion with blind puncture is safe and effective.

Objective To sum up the experience in the diagnosis and management of acute lower extremity arterial injury.Methods Between Jan 1988 and Feb 2004,125 cases of lower limb arterial injuries were admitted and undergoing surgery, including gunshot in 3 cases, blunt trauma in 56 and stabbing in 66 cases. Associated injuries included bony injury in 36 cases, nerve injury in 11 cases, and vein injury in 23 cases. Arterial primary or patch repair was performed in 28 cases, end-to-end anastomosis in 37 cases, saphenous vein graft used in 32 cases, prosthetic graft bypass in 23 cases, thrombectomy in 2 cases and blood vessel ligation in 3 cases.Results Limb salvage rate was 85.6% and patient survival of 98.4%. Massive bleeding and multiple organ failure caused mortality in one each. The rate of amputation is 10.4%(13/125), with preoperative gangrene being present in 8 cases.Conclusion Lower extremity arterial injury carries a high amputation rate. The use of Doppler scanner is helpful for early diagnosis. An ankle/brachial index of less than 1 in the affected limb was considered as a positive sign of arterial injury. Prompt revascularization and early fasciectomy are important to reduce amputation rate and mortality.

OBJECTIVETo investigate the potential developmental toxicity of Radix Ophiopogonis decoction in SD rats.

METHODTimed-pregnant SD rats were given Radix Ophiopogonis decoction (26.9 g x kg(-1)) or vehicle (distilled water) by gavage on gestation days 6-17. Maternal clinical sign, abortions, premature deliveries, and body weight were monitored throughout gestation. At termination (gestation days 20) pregnant females were evaluated for clinical status and gestational outcome; live fetuses were examined for gender, external, visceral and skeletal malformation and variations.

RESULTNo deaths, premature deliveries or dose-related clinical signs were attributed to Radix Ophiopogonis decoction. Maternal body weight and body weight gain were not affected. There were no effects on fetus weight and viability, incidences of fetal malformation and variation.

CONCLUSIONThese results demonstrated that Radix Ophiopogonis decoction had no detectable adverse effects in either the treated F0 female rats or the fetuses.

Animals ; Body Weight ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; adverse effects ; Embryonic Development ; drug effects ; Female ; Fetal Development ; drug effects ; Fetal Weight ; drug effects ; Humans ; Male ; Models, Animal ; Ophiopogon ; chemistry ; Pregnancy ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the perioperative and long-term effects of endovascular aneurysm repair(EVAR) of infrarenal abdominal aortic aneurysm (AAA).Methods Clinical data of 131 AAA cases undergoing EVAR were retrospectively evaluated for the safety and long-term efficacy.Results The operative time was (137 ±29) min,blood loss was (142 ±20) ml,blood transfusion was (46 ± 26) ml,ICU staying time was (17 ± 4) h.Major perioperative complications were severe heart failure in 8 cases,myocardial infarction in 2 cases,pulmonary complications in 5 cases,internal leakage in 4 cases.During the period of up to 60 months there were15 cases of endoleak including 8 cases of type Ⅰ,5 cases of type Ⅱ,1 each case of type Ⅲ and Ⅳ and 2 deaths.By Kaplan-Meier survival analysis there were complications developing after 60 months and up to 40％ of them needing reintervention.Conclusions Endovascular repair is the safe treatment for AAA,but discharged patients need close long-term follow-up.Complications that ensued need intensive management.