Fibrolase is a non-hemorrhagic zinc metalloproteinase isolated from southern copperhead snake venom (Agkistrodon contortrix contortrix) and is capable of degrading fibrin clots resulting from purified fibrinogen or from blood plasma. Alfimeprase, a truncated form of fibrolase, as a clinical agent was successfully completed PhaseII clinical trials.The cDNA of alfimeprase was amplified by recursive PCR, digested with BamHI and HindII, and cloned into pET43.1a, pMALp2X and pMALc2X vectors to generate fusions with NusA, MBP and sMBP(with signal peptide), respectively. Nus/alfimeprase was expressed in soluble form by co-expressing with chaperone FkpA and inducing with1mmol/L IPTG. The fusion protein accounted for about 25 % of total protein following cell lysis. Alfimeprase was successfully purifiesd by Ni-NTA affinity chromatography and cleaved by enterokinase. The results demonstrate the fibrinolytic activity of recombinant alfimeprase using fibrin plate assays and fibrinogen hydrolysis.

Bridged-Ring Compounds ; poisoning ; Humans ; Male ; Middle Aged ; Poisoning ; therapy ; Treatment Outcome

Objective:To compare the release ability of water-soluble polysaccharides in sporoderm-broken and nonbroken spores of Ganoderma lucidum, and establish a comparatively correct method for the determination and analysis of polysaccharide contents in Chinese herbs.Method:The release ability of water-soluble polysaccharides was determined on the basis of phenol-sulfuric acid modification in different conditions.Result:The release ability of polysaccharides of sporoderm-broken spores was much grerter than that of nonbroken spores; and the phenol-sulfuric acid modified cation method proved excellent in accuracy and reproducibility, with a relative error less than 1.5%.Conclusion:The spores should be wall-wracked if used as a nutriment, or for extraction and analysis of their effective components. The method can be successfully used for the determination of polysaccharide contents in Chinese herbs or nutriments.

Objective To study the level of serum lipids and body-fat content of high-fat diet induced pbesity rats(DIO).explore the relationship between intracellular caleium and ventrieufar arrhythmia.Methods Sixty male Sprague-Dawley(SD)rats were divided into control group(15)and experiment group(45),high-fat diet was administrated for 12 weeks to established obesity model,15 rats were selected into obesity group according to their body weight gain.The standard 2-lead electrocardiograph was used to detect the incidence and scores of arrhythmia induced by barium chloride(BaCl2,0.1 mg/kg)for 1 hour on every 8 rats from different groups respectively.Body-fat content.the level of serum total cholesterol(TC),triglyceride(TG),low density lipopmtein cholesterol (LDL-C) and high density lipopmtein cholesterol (HDL-C)were measured.The epididymal(EP),retroperitoneal (RE) and mesenteric(ME)white adipose pads was measured to obtain the body fat content.Single ventricular myrocytes of rats were isolated by enzymatic dissociation.The confocal laser scanning microscope was used to record basic intraceUular calcium level([Ca2+]i).Results The body-fat content in obesity group[(7.71±0.74)%] was significantly higher than control group[(4.69±0.37)%](t＝3.650,P＜0.05).The level of serum TC,TGand LDL-C were significantly higher(t＝3.801,2.778,3.536.P＜0.05) in obesity group[(1.26±0.04),(0.58±0.10),(0.51±0.04)mmol/L]than those in control group[(0.92±0.08),(0.29+0.03),(0.31±0.04) mmol/L].The level of serum HDL-C wa8 decreased gradually from control group[(0.53±0.05)mmol/L] toobesity group[(0.52±0.02)mmol/L],but there waft no significant difference between them(t＝0.186,P＞0.05).The incidence of arrhythmia induced by BaCl2(0.1 mg/kg)in obesity group was significantly higher than controlgroup(X2＝5.333,P＜0.05),and the scores of arrhythmla was increased in obesity group(2.5±0.6)too.The fluorescence intensity standing for[Ca2+]i was increased significantly(t＝2.409,P＜0.05)from obesity group(247.96±20.03)to control group (174.25±23.13).Conclusion As the free cytosolic calcium begin to accumulate,the arrhythmia morbidity is increased in obesity rats.

OBJECTIVETo explore the feasibility of using an intracavitary convex array probe for detecting the distal extracranial internal carotid artery (ICA) by transoral carotid ultrasonography (TOCU).

METHODSForty patients underwent examinations with bilateral ICA inspected with an intracavitary convex array probe by TOCU to observe the internal diameter, visible length, peak systolic velocity (PSV), end-diastolic velocity (EDV) and resistance index (RI).

RESULTSEight of the 40 patients were excluded from the observation for the presence of carotid plaques. The examination was terminated in two patients due to sensitive throat and severe pharyngeal reflex. The rest of the patients completed the examination of the internal diameter, visible length, PSV, EDV and RI, which showed no statistically significant differences among them (P>0.05).

CONCLUSIONUsing intracavitary convex array probe, the distal extracranial ICA disease can be diagnosed with higher accuracy.

Adult ; Aged ; Carotid Artery, Internal ; diagnostic imaging ; Female ; Humans ; Image Processing, Computer-Assisted ; methods ; Male ; Middle Aged ; Mouth ; diagnostic imaging ; Ultrasonography, Doppler, Duplex ; methods

OBJECTIVETo investigate the therapeutic results after various kinds of partial laryngectomy for patients with glottic carcinoma of the larynx.

METHODSFrom January 1980 to Mar 2004, 98 patients with glottic carcinoma of the larynx were treated by various kinds of partial laryngectomy. The types of operation varied according to stage of the lesion: vertical or frontovertical partial laryngectomy for T1b, T2 and T1a with invasion of anterior commissura, Tuker's operation or supracricoid partial laryngectomy for T1b, T2 or those with invasion of over half of contralateral cord, and extended vertical partial laryngectomy with resection of arytenoids for T3, or with invasion of ipsilateral arytenoids cartilage without involving postcricoid mucosa or posterior commissura. Totally, vertical partial laryngectomy was carried out for 76 patients, extended vertical partial laryngectomy for 7, supracricoid partial laryngectomy for 3 and Tucker's operation for 12. One stage unilateral neck dissection was performed in 5 patients and postoperative radio-therapy was supplemented to 5 (dose 60 Gy). For laryngeal function restoration, pectoro-hyomyo-flap and cricohyoidoepi glotopexy procedures were also performed for some patients. The clinical data of these patients were retrospectively analyzed.

RESULTSThe 3- and 5-year survival rate was 86.5% (64/74) and 81.7% (49/60), respectively. The total decannulation rate was 94.9% (93/98), though fifteen patients had had symptoms of aspiration. Swallowing function of all patients recovered to the normal level. The phonation of all patients was restored to various degrees and they were able to communicate socially.

CONCLUSIONThe 3- and 5-year survival rates of partial laryngectomy for patients with glottic carcinoma of the larynx are comparable to the results achieved by total laryngectomy, but the quality of life of the patents much better. Complete resection of the tumor and simultaneous preservation of respiratory and vocal function whenever possible by partial laryngectomy is very important for improving patients' life quality. With a suitable size and good blood supply, the sternohyoid muscle flap of unipedical and half-thickness instead of whole thickness is ideal in the reconstruction of laryngeal defect after vertical partial laryngectomy.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; surgery ; Female ; Glottis ; surgery ; Humans ; Laryngeal Neoplasms ; surgery ; Laryngectomy ; methods ; Male ; Middle Aged ; Quality of Life ; Retrospective Studies ; Treatment Outcome

Objective To develop a supporting pole for medical detachable camouflage net at field conditions to enhance the timeliness of battlefield camouflage of deployable units such as medical tent.Methods The pole had a dumbbell-shaped,hollow and columnar structure,which was composed of a base,a pole body and a terminal disc.The pole body consisted of internal and external parts.The external part had a vertical opening at its top and screw thread at its side wall,which was equipped with a binding bolt.The terminal disc had a circular structure and a 20 cm outer diameter,which was fixed 10 cm under the top of the internal pole to support the net.The base had a center hole and a sleeve to hold the lower part of the external pole to immobilize the supporting pole.Results The supporting pole decreased the deployment time of the camouflage net from 5 min to 2 min and the withdrawal time from 4 min to 1.5 min,and enhanced the timeliness of all-element deployment of mobile medical unit during field practical training.Conclusion The supporting pole meets the tactical requirements for deployment,storage,transport and robustness,and thus is worthy promoting in medical unit.

Adult ; Hematoma, Epidural, Spinal ; etiology ; Humans ; Male ; Spinal Fractures ; complications ; surgery ; Thoracic Vertebrae ; injuries ; surgery

Hepatitis B, Chronic ; physiopathology ; psychology ; Humans ; Liver ; physiopathology ; Stress, Psychological ; physiopathology

OBJECTIVETo prepare macroporous bone substitute composed of calcium phosphate cements and rhBMP-2 loaded gelatin microspheres, and to investigate ectopic osteoinduction of the composite.

METHODSAfter being prepared by improved emulsified cold-condensation method and crosslinked by 5% genipin solution,gelatin microspheres (GMs) were observed by scanning electron microscope (SEM) and loaded with rhBMP-2 by adsorption. Macroporous bone substitute was developed by mixing calcium phosphate cement (CPC) with 2.5% GMs, being as the experimental group,and CPC with rhBMP-2 was the control group. After the both composites had been soaked in the sodium chloride for 1 week or 3 weeks, compressive strength of the composites were tested, and the cross-sections were observed by SEM. Concentrations of rhBMP-2 in the solutions at different time by ELISA method and the cumulative drug release amount was calculated. The composites had been implanted in the muscle bags of the mouses for 3 weeks. Then the tissues around the materials were collected, stained by hematoxylin and eosin, and Ca and ALP in the tissues were also measured.

RESULTSGelatin microspheres were spherical with diameters of (62 +/- 18) microm. Macropores appeared in the experimental materials 1 week and 3 weeks after being soaked,and total porosity, macroporosity, cumulative release amount of rhBMP-2 in the experimental group were higher than that in the control. But compressive strength of the experimental group was lower than that of the control group 3 weeks after being soaked. Results of HE stain showed chondral formation in both groups, but there were more chondral tissues in the experiment group, and so were the concentrations of Ca and ALP.

CONCLUSIONMacroporous calcium phosphate cement can be prepared by using rhBMP-2 loaded gelatin microspheres, and it is an excellent bone substitute due to it's proterty of promoting rhBMP release and powerful ectopic osteoinduction.

Animals ; Bone Cements ; chemistry ; Bone Morphogenetic Protein 2 ; chemistry ; Bone Substitutes ; chemistry ; pharmacology ; Calcium Phosphates ; chemistry ; Gelatin ; chemistry ; Humans ; Male ; Mice ; Microspheres ; Osteogenesis ; drug effects ; Porosity ; Recombinant Proteins ; chemistry