Efforts on directed evolution of sortase A to optimize its catalytic properties have been undertaken and shown the promise. To facilitate screening of sortase A mutants with expected catalytic properties, a novel ligation efficiency monitoring system, including reporter substrates GFP-LPETG and GGGYK-Biotin, was developed. GFP-LPETG, wild type sortase A, and a recently reported high activity sortase A mutant were prepared recombinantly from Escherichia coli BL21 (DE3). Taking advantage of the newly designed reporter system, the ligation efficiency catalyzed by wild type and mutant form of sortase A could be sensitively monitored via SDS-PAGE directly. Consistent with previous report, the mutant sortase A displayed much higher catalytic activity compared to wild type enzyme, indicating the new reporter system is easily and fast handled and sensitive. The application of this reporter system into systemic screening will facilitate future directed optimization of sortase A.
Aminoacyltransferases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Biocatalysis ; Biotin ; Cysteine Endopeptidases ; genetics ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; Genes, Reporter ; Ligation ; Mutant Proteins ; genetics ; metabolism

Adult ; Female ; Heart Valve Prosthesis ; Heart Valve Prosthesis Implantation ; adverse effects ; Humans ; Tachycardia, Reciprocating ; etiology ; Tachycardia, Ventricular ; etiology

Seven meroterpenoids and five small-molecular precursors were isolated from Penicillium sp., an endophytic fungus from Dysosma versipellis. The structures of new compounds, 11beta-acetoxyisoaustinone (1) and isoberkedienolactone (2) were elucidated based on analysis of the spectral data, and the absolute configuration of 2 was established by TDDFT ECD calculation with satisfactory match to its experimental ECD data. Meroterpenoids originated tetraketide and pentaketide precursors, resepectively, were found to be simultaneously produced in specific fungus of Penicillium species. These compounds showed weak cytotoxicity in vitro against HCT-116, HepG2, BGC-823, NCI-H1650, and A2780 cell lines with IC 50 > 10 micromol x L(-1).
Berberidaceae ; microbiology ; Cell Line ; Cell Line, Tumor ; Humans ; Lactones ; isolation & purification ; pharmacology ; Monoterpenes ; isolation & purification ; pharmacology ; Penicillium ; chemistry

BACKGROUNDPercutaneous microballoon compression (PMC) for trigeminal neuralgia is an important therapeutic method. The aim of this study was to review the effects of PMC for trigeminal neuralgia in 276 patients.

METHODSFrom December 2000 to May 2003, 276 patients with trigeminal neuralgia were treated with PMC. The course of the disease ranged from 3 months to 38 years. Under the guidance of C-arm X-ray, 14# needle was placed into the foramen ovale using the classical Hakanson's technique. Fogarty balloon catheter was navigated into the Meckel's cave tenderly. A small amount of Omnipaque was slowly injected to inflate the balloon and compress the trigeminal ganglion for 3 to 10 minutes.

RESULTSA total of 290 PMC were performed on the 276 patients. Among them, 252 had immediate relief from pain. The patients were followed up for a mean of 18.7 months (range, 4 to 32), 14 of them had a recurrence. Of the 14 patients, 12 were re-operated with PMC, and the pain was all controlled successfully.

CONCLUSIONSPMC is an effective and technically simple method for trigeminal neuralgia. For older patients with trigeminal neuralgia, it may be the first choice.

Adult ; Aged ; Aged, 80 and over ; Blood Pressure ; Catheterization ; methods ; Female ; Follow-Up Studies ; Heart Rate ; Humans ; Male ; Middle Aged ; Trigeminal Neuralgia ; physiopathology ; therapy

OBJECTIVE: To provide anatomic data of pterygopalatine fossa(PPF) for endoscopic PPF surgery. METHOD: Fifteen wet adult skull specimen fixed in 10% formaldehyde were studies. From center the skulls were sawed, the middle and inferior turbinate were resected, the anterior and posterior ethmoid sinuses were resected. Then after punch out perpendicular part of palatine bone and posterior wall of maxillary,the pterygopalatine fossa were showed and some related measurements were made. RESULT: This pathway can lead to a good view of pterygopalatine fossa. The main soft structures in PPF were maxillary artery and nerve with their branches. The diameter of artery branches was smaller than 3 mm. CONCLUSION The endoscopic PPF surgery is safe and practical from the anatomic data.
Adult ; Endoscopy ; Ethmoid Sinus ; anatomy & histology ; Humans ; Maxilla ; anatomy & histology ; Maxillary Artery ; anatomy & histology ; Maxillary Sinus ; anatomy & histology ; surgery ; Nasal Cavity ; surgery ; Palate, Hard ; anatomy & histology ; Pterygopalatine Fossa ; anatomy & histology ; surgery

Objective To evaluate the methods and consequences of surgical technique in the treatment of Stanford A aortic dissection.Methods 108 patients with type Standford A aortic dissection underwent surgery in our study,including urgent surgery in 53 and selective surgery in 55.The operation was performed under deep hypothennic circulatory arrest (DHCA) in 85 cases.Surgical procedures included ascending and semi arch replacement or total arch replacement (some cases combined with stented graft implanted into the descending aorta),"elephant trunk" procedure.Concomitant procedures included repair of intimal tear in arch or descending aorta,Bentall procedure,aortic valve replacement,Cabrol or modified Cabrol procedure,aortic valvuloplasty,mitral valvuloplasty or mitral valve replacement,tricuspid valvuloplasty and CABG.Results In-hospital mortality was 6.5% (7 of 108 patients).The mor- tality was 7.5% (4 of 53 patients) in urgent surgery group and in elective surgery group was 5.4% (3 of 55 patients).Ninety six percent survived patients were followed up for 1 month to 13.3 years [mean (3.2?1.3) years] and 2 deaths occurred during the fel- low-up period.3 patients underwent re-operatian.Conclusion The choice of surgical procedures depend on the location of intimal tear for Stanford A aortic dissection.The better operative effects can be expected with proper surgical indication,perfecting surgical technique,and enhancing postoperative treatment.

BACKGROUNDPremature ventricular contraction (PVC) is one of the most common kinds of arrhythmias for which the treatment falls into dilemma. Previous clinical application showed that the traditional Chinese Medicine Shensongyangxin (SSYX) capsule is efficacious for the treatment of PVCs. This randomized clinical trial aimed to further evaluate the efficacy and safety of SSYX capsule on treating PVC.

METHODSThe subjects who had frequent PVCs with or without organic heart disease and normal cardiac function were enrolled in the study. The primary endpoint was the change of PVC numbers after eight-week medication with SSYX capsule. The secondary endpoints included change of clinical symptoms related to PVCs and the safety evaluation of SSYX capsule. Totally 188 PVC patients were randomly enrolled in the non-organic heart disease PVCs trial and orally took either SSYX capsules or analogues (three times per day, 4 capsules one time). A total of 671 PVCs patients were randomly enrolled in the organic heart disease PVCs trial, and orally took either SSYX capsules (three times per day, 4 capsules one time) or mexiletine tablet (three times per day, 150 mg one time). The PVCs were monitored and calculated with 24-hour Holter electrocardiogram. Routine blood, liver and kidney function were tested before and after medication with SSYX capsule.

RESULTSSSYX capsules significantly decreased the PVCs numbers and alleviated the related symptoms in patients with or without organic heart disease. In non-organic heart disease group, SSYX capsules and the placebos decreased the PVCs from 12,561.34 ± 9,777.93 to 4,806.87 ± 6,507.17, and 12,605.69 ± 8,736.34 to 10,364.94 ± 9,903.41, respectively. The total effective rate was 74.2% and 28.9% in SSYX and placebo groups (P < 0.001). In organic heart disease group, SSYX capsule and mexiletine decreased the PVCs from 8,641.01 ± 8,923.57 to 3,853.68 ± 7,096.42, 8,621.61 ± 8,367.74 to 5,648.29 ± 8,667.38, respectively. The total effective rate was 65.8% and 50.7% in SSYX and mexiletine groups (P < 0.001). In addition, SSYX capsule significantly alleviated PVCs-related symptoms such as palpitations, chest tightness, insomnia, fatigue, and night sweats. No adverse cardiac events were observed except some slight gastrointestinal side effects during the study.

CONCLUSIONSCompared with placebo or mexiletine, SSYX capsules have significant therapeutic efficacy in reducing PVCs numbers and alleviate PVCs-related symptoms.

Capsules ; therapeutic use ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; Ventricular Premature Complexes ; drug therapy

OBJECTIVETo investigate the distribution and drug resistance of acinetobacter isolated from burn wounds.

METHODSThe acinetobacter strains were isolated and identified by routine methods. Based on the recommendation of NCCLS, AmpC enzyme was determined by cefoxiti three-dimensional test, ESBLs by disk diffusion method and bacterial susceptibility by Kirby-Bauer agar diffusion method.

RESULTSAmong the 69 strains of acinetobacter clinically isolated from burn wounds, 52 were baumannii (75.6%). The acinetobacter strains were identified to be highly resistant to 17 kinds of antibiotics. The drug resistance rate of beta-lactamase-producing strains (68.25%) was higher than that of non-beta-lactamase-producing strains (20.33%). The strains isolated in our burn ward exhibited multiple drug resistance which was mainly due to the production of many kinds of beta-lactamases. Among the 38 strains of beta-lactamase-producing acinetobacter, those producing AmpC beta-lactamase (AmpC BLA) accounted for 42.1%.

CONCLUSIONAcinetobacter strain was one of the pathogens in burn wound infection, and its isolation and identification of its drug resistance could be beneficial to the doctors to make right choice of antibiotics.

Acinetobacter ; classification ; enzymology ; isolation & purification ; Anti-Bacterial Agents ; pharmacology ; Burn Units ; Burns ; microbiology ; Drug Resistance, Bacterial ; drug effects ; Drug Resistance, Multiple ; drug effects ; Humans ; Membrane Proteins ; metabolism ; Ribosomal Proteins ; metabolism

Acetylcholine ; metabolism ; Animals ; Arcuate Nucleus of Hypothalamus ; metabolism ; Dopamine ; metabolism ; Epinephrine ; metabolism ; Female ; Neurotransmitter Agents ; metabolism ; Norepinephrine ; metabolism ; Physical Conditioning, Animal ; Rats ; Rats, Sprague-Dawley ; Synaptic Transmission

Objective To establish the animal model for the study of children with moderate blood lead levels in young rabbits,for the study of the ideal therapy for moderate lead poisoning in children.Methods Sixteen 45-day-old male New Zealand rabbits were randomly divided into control and lead-exposed group,8 in each group.Rabbits in the lead-exposed group were treated with 5 mg/(kg?d)lead acetate in their forage for 6 weeks to establish moderate lead poisoning animal model.The blood lead levels(BLLs)were determined by atomic absorption spectrometer(AAS),and the urine lead levels and the lead concentrations of tissue and organ were determined by inductively coupled plasma-mass spectrometry(ICP-MS).Histopathology in tissue and organ was observed under the light microscope.Results The BLLs and the urine lead levels in lead-exposed group step up rapidly in primal weeks,then retained at a steady levels.The BLLs exhibited moderate level BLLs during the lead exposure period.Compared with control group,the body weight gain,testis and hippocampus wet coefficient of the lead-exposed group significantly decreased(P_a