The structural gene encoding mature peptide of extracellular ? amylase was amplified from the genome DNA of hyperthermophilic archaeon Pyrococcus furiosus by PCR The recombinant plasmid pUC19 amy was constructed by inserting the amplified segment into vector pUC19 The recombinant vector pYX212 amy was constructed by ligate the heterogeneous fragment of pUC19 amy into the multiple cloning site of pYX212, an expression vector of yeast Saccharomyces cerevisiae W303 A1 were transformed with pYX212 amy by electroporation The transformant expressed the activity of the thermophilic ? amylase successfully The recombinant enzyme has the similar enzymatic properties as the extracellular ? amylase produced by Pyrococcus furiosus : it shows an enzymatic activity optimum at pH 5 0, and its optimal temperature for enzymatic activity is about 90℃, more than 50% of its initial enzymatic activity is still detectable after it was incubated at 121℃ for 30 minutes

Objective To evaluate the efficacy of gentamicin solution in transurethral ureteroscopic lithotripsy to prevent postop- erative infection.Methods Prospective clinical randomized control study was conducted.From July 2003 to June 2006,116 ca- ses of ureteral stones at high risk of postoperative infection were randomized into control group or gentamicin group.Patients in gentamicin group received gentamicin solution for washing in the operation.All the patients undergoing operation were followed up for 2 weeks after operation.Diagnosis of postoperative infection was based on clinical manifestations.Results A total of 109 patients received operation in all the 116 cases,including 58 cases in gentamicin group and 51 cases in control group.Thirteen cases of postoperative infection were identified in all the patients receiving operation (11.93%),3 cases in gentamicin group and 10 in control group.The incidence of postoperative infection was significantly different between the two groups (X~2= 5.3342,P=0.0209).Eight cases had positive bacterial culture.Of the microbiological isolates,2 were gram-positive bacteria, 5 gram-negative bacteria and 1 Candida albicans.Conclusions The most common pathogen causing postoperative infection after transurethral ureteroscopic lithotripsy is gram-negative bacteria.The use of gentamicin solution for washing in the operation can reduce the incidence of postoperative infection.

Biomass is a renewable resource, which can be transformed into useful chemical products. The effects of dilute hydrochloric acid on the hydrolysis of steam explosion pretreatment of corn straw were studied. This article developed the application research of ergosterol using corn straw hydrolysates as fer- mentation substrates. The results showed that when corn straw was hydrolyzed with 1.5% hydrochloric acid, temperature at 90?C, hydrolysis for 3 h and the corresponding solid to liquid ratio at 10%, the reducing sugar content can reached up to 53.3% and cellulose conversion efficiency was 79%. The optimal fermental pa- rameters were as follows: 6.0 oBx of corn straw hydrolysates, corn concentration steep water at 4%, pH 7.5, 10% of inoculation, 28?C cultivated for 32 h. Under these conditions, the yeast biomass up to 8.5 g/L and the ergosterol content up to 2.35%. The infrared spectrometer and the X-ray diffract meter used to characterize of crystallite structure.

Objective To quantitatively analyze the coronary artery ostia by three-dimensional trans-esophageal echocardiography (3D-TEE).Methods The full-volume images of aortic root and coronary artery ostia were acquired by 3D-TEE in 95 adult patients.The Philips QLab 3DQ measurement technology was employed to determine three mutually perpendicular planes:(1) The transverse plane cross the bottom of three coronary artery sinus.(2) The sagittal plane perpendicular to sino-tubular junction.(3) The coronal plane perpendicular to the aforementioned two planes .The following relevant parameters were measured and recorded:(1) Length, width, height and area of bilateral coronary artery ostia .(2) The angle between coronary arterial outflow tract and aortic root in sagittal plane .(3) The spatial distribution of coronary artery ostia, aortic root and coronary artert sinus .Results The shape of left coronary artery ostia were more regular (round or oval) than right coronary artery ostia ( teardrop-shape or oval ).Calcification was more frequent in right coronary artery ostia (81/95, 85.26%) than that in left coronary artery ostia. There were statistical differences between left and right coronary artery in the parameters of ostial wide , area and height (t =3.85, 3.86, -4.49, all P<0.01).Most left coronary artery ostia were located inside the sinus (76/95, 80.00%), mainly in the upper third segment (69/95, 72.63%); while more than half of the right coronary artery ostia were found outside the sinus ( 53/95, 55.79%).The difference was statistically significant( χ2 =25.91, P<0.01).Conclusion The quantitative analysis of aortic root and coronary artery ostia based on the full-volume images originated from real-time 3D-TEE is feasible, which is helpful for further clinical research .

Aged ; Astragalus Plant ; Cerebral Infarction ; drug therapy ; Drug Combinations ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phytotherapy ; Plant Extracts ; Platelet Aggregation ; drug effects ; Salvia miltiorrhiza

Acute Disease ; Adult ; Humans ; Male ; Occupational Diseases ; diagnosis ; therapy ; Phosphines ; poisoning ; Poisoning ; diagnosis ; therapy

Objectives: To evaluate the clinical significance of nuclear matrix protein 22 (NMP 22) in the detection of bladder transitional cell carcinoma (BTCC) and compare with voided urine cytology(VUC). Methods: A total of 69 cases with voided urine samples for NMP 22 and VUC test were included in this study. Thirty of them were BTCC patients(BTCC group) and twenty nine suffered from other urological diseases (nonbladder cancer group, NBC group). Ten were healthy volunteers (control group). Results: The NMP 22 values for BTCC group (67.3 U/ml) were significantly higher than that of NBC group(7.4 U/ml) and control group (4.3 U/ml)( P 0.05). NMP 22 was more sensitive than VUC in low grade BTCC(Ⅰ,Ⅱ)(62.50% vs 12.50%,P 0.05). Conclusions:Urinary NMP 22 is a useful marker for the early diagnosis of BTCC. It is more sensitive than VUC in low stage and grade BTCC.

Objective: To investigate the inhibitory effect of adenovirus-mediated VEGF-siRNA on transplanted osteo- sarcoma in nude mice.Methods: VEGF-siRNA gene was cloned into the genome of replication-deficient adenovirus to construct Ad-VEGF-siRNA;the latter was then used to infect osteosarcoma MG63 cell line in vitro;and the expression of VEGF gene was detected by RT-PCR.Osteosarcoma transplantation model was established in nude mice;VEGF expression in tumor tissue was analyzed and the inhibitory effect on tumor growth and lung metastasis were also observed.Results: The recombinant adenovirus vector Ad-VEGF-siRNA was successfully constructed.In vivo and in vitro experiment both showed that Ad-VEGF-siRNA significantly downregulated VEGF expression in MG63 cells and transplanted tumor tissue. It was found that Ad-VEGF-siRNA significantly inhibited transplanted osteosarcoma growth(P

The ergosterols were produced from corn straw hydrolysates fermented by ergosterol yeast,which was obtained from protoplast electrofusion.The effects on the yield of ergosterol were studied in the condition of shaker,such as initial sugar concentration,nitrogen source,pH value and fermentation time.The technical conditions were optimized according to the DPS center-united experimental design principles and the method of response surface analysis with four factors and three levels.The results indicated that the four factors had significant correlation to ergosterol accumulation.The biomass and the ergosterol content could be up to 8.67g/L and 2.37% respectively after cultivated for 32h under optimal technical condition.The structure of ergosterol crystal was characterized by UV,IR and SEM.A new approach of biomass source application was presented.

Objective To induce mouse embryonic stem(ES)cells to differentiate into insulin-secreting cells by means of a 5-step model system.Methods E14.1 mouse ES cells were cultured in the presence of leukemia inhibitory factor(LIF)for 2 days(step 1),then the cells were cultured in hanging drops to form embryonic bodies(EBs)and the resulting EBs were cultured in suspension for 6 days in the presence of basic fibroblast growth factor bFGF(step 2).Subsequently the EBs were cultured in the medium containing glucagon- like peptide 1(GLP-1),hepatocyte growth factor(HGF),nerve growth factor(NGF)and nicotinamide for 10 days(step 3).After that,the EBs were dissociated into single cells,and the cells were cultured in monolayer in the presence of GLP-1,betacellulin,activin A,bFGF and nicotinamide for 10 days(step 4).Finally,the cells were cultured in low-glucose medium containing nicotinamide for 4 days(step 5).Insulin and some other islet- related genes expressions were investigated using RT-PCR and insulin expression was also investigated by DTZ- staining and immunohistochemistry.The percentage of insulin-secreting cells was evaluated by flowcytometry and insulin concentrations were measured by RIA.Results mRNA expression of insulin became visible at step 3 and more evident at step 5.Additionally,at step 5,mRNAs of glucagon,somatostatin,pancreatic polypeptide(PP), pancreatic duodenal homeobox 1(PDX-1),beta-cell E box transactivator 2(Beta2)and neurogenin 3(Ngn3) were detected.DTZ-staining positive cells and insulin immunohistochemical staining positive cells were observed. The percentage of insulin-positive cells was(24.0?2.5)%(n=6).In the presence of 5.6 mmol/L and 25 mmol/L glucose,insulin concentrations were(0.05?0.01)?g/L and(0.13?0.02)?g/L respectively(n= 6).Conclusion E14.1 mouse ES cells can be induced to differentiate into insulin-secreting cells by the 5-step model system.Insulin-secreting cells can release insulin into culture medium when treated with glucose,and insulin concentrations increase with rising concentration of glucose.