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

Cellulosic material is the most abundant renewable carbon source in the world. Cellulose may be hydrolyzed using cellulase to produce glucose, which can be used for production of ethanol, organic acids, and other chemicals. Cellulase is a complex enzyme containing endoglucanase (EC 3.2.1.4), exoglucanase (EC 3.2.1.91) and cellobiase (EC 3.2.1.21). The hydrolysis of natural cellulose to glucose depends on the synergism of these three components. The mostly used cellulase produced by Trichoderma reesei has high activity of endoglucanase and exoglucanase, but the activity of cellobiase is relatively low. Therefore, improving the activity of cellobiase in cellulase reaction system is the key to enhance the sacchrification yield of cellulosic resources. Aspergillus niger LORRE 012 was a high productivity strain for cellobiase production. It was found that the spores of this strain were rich in cellobiase. In this work, the cellobiase was immobilized efficiently by simply entrapping the spores into calcium alginate gels instead of immobilizing the pure cellobiase proteins. The immobilized cellobiase was quite stable, and its half-life was 38 days under pH 4.8, 50 degrees C. The thermal stability of the immobilized cellobiase was improved, and it was stable below 70 degrees C. The suitable pH range of the immobilized cellobiase was pH 3.0 - 5.0, with the optimal pH value 4.8. The Km and Vmax value of the immobilized cellobiase were 6.01 mmol/L and 7.06 mmol/min x L, respectively. In repeated batch hydrolysis processes, 50 mL of substrate (10 g/L cellobiose) and 10 mL of immobilized beads containing cellobiase were added into a 150 ml flask. After reacting at pH 4.8, 50 degrees C for several hours, the hydrolysate was harvested for assay, and the immobilized beads were used for the next batch hydrolysis with the fresh substrate. This process was repeated, and the yield of enzymatic hydrolysis kept higher than 97% during 10 batches. The continuous hydrolysis process was carried out in a column reactor (inside diameter 2.8 cm, inside height 40 cm) packed with the immobilized beads. Using 10 g/L cellobiose as substrate, the hydrolysis yield reached 98% under 0.4 h (-1) dilution rate and pH 4.8, 50 degrees C. After corncob was treated by 1% dilute acid, the cellulosic residue (100 g/L) was used as substrate, and hydrolyzed by the cellulase (15 IFPU/g substrate) from Trichoderma reesei, at pH 4.8, 50 degrees C for 48 h. The concentration of reducing sugar in the hydrolysate was only 48.50 g/L (hydrolysis yield 69.5%). When the hydrolysate was further treated by the immobilized cellobiase, the cellobiose was hydrolyzed into glucose, and the feedback inhibition caused by the cellobiose accumulation disappeared sharply. By the synergism of immobilized cellobiase and the cellulase from T. reesei left in the hydrolysate, other oligosaccharides were mostly converted to monosaccharides. At 48 h, the reducing sugar concentration was increased to 58.78 g/L, the hydrolysis yield of the corncob residue was improved to 84.2%, and the ratio of the glucose in the total reducing sugar was increased from 53.6% to 89.5%. The reducing sugars converted from corncob could be used further in the fermentation of valuable industrial products. This research results were meaningful in the conversion and utilization of renewable biomass.
Aspergillus niger ; enzymology ; Biotechnology ; Cellobiose ; metabolism ; Enzyme Stability ; Hydrogen-Ion Concentration ; Kinetics ; Temperature ; beta-Glucosidase ; chemistry ; metabolism

Objective To understand the evaluation on graduates' professional quality from the employment institutions,and to provide the references for the higher vocational schools in improving students' training model and strengthening students' professional quality. Methods The questionnaires designed independently were answered by the employment institutions hiring 2003-2005 four majors(namely nursing,laboratory science,dental technology,optometry) graduates from Shanghai Institute of Health Sciences. Results The overall evaluation of employment institutions on the graduates was quite good,but the evaluation on graduates' professional ability was low. Conclusion With strengthening the student' specialized knowledge and skill training,higher vocational schools must pay more attention to the students' professional ability education.

Objective To observe the effect of atorvastatin on mixed dyslipidemia in type 2 diabetic patients. Methods 39 patients of type 2 diabetic with mixed dyslipidemia were taken with atorvastatin for 6 months,and the change of total cholesterel(TC),triglyceride(TG),low-debsity-lipoprotein-cholesteral(LDL-C),high-debsity-lipopro- tein-cholesteral(HDL-C)were observed.The incidence of side-effect was recorded.Results TC,TG,LDL-C were obviously reduced and their extent of reduction were 26.17 %,54.97 %,38.92 %.HDL-C was obviously increased and it's extent of increase was 14.81%(P

The ?glutamyltranspeptidase encoding gene(ggt) from Bacillus subtilis SYU 20016 was amplified by PCR. The ggt gene was inserted in pBV220 to yield the recombinant expression vector pBV220ggt. Overexpression of ggt in E.coli JM109 was achieved with pBV220ggt. SDSPAGE analysis showed an overexpressed recombinant product at about 65kDa,consistent with the molecular weight predicted from gene sequence. The ferment conditions of r-glutamyltranspeptidase were also discussed. The optimum temperature and pH for the enzyme were determined as 30℃ and 7.2 respectively.The cultures were incubated at 42℃ for 4h with broth volume 20ml/250ml flask and the yield of 6U/ml was obtained, enzyme activity of B. subtilis NX2 was only 3.2 U/ml.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Liver Neoplasms, Experimental ; drug therapy ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Metastasis ; prevention & control ; Neoplasm Transplantation ; Phytotherapy ; Qi ; Spleen

OBJECTIVETo determine the effects of Jiji decoction (Traditional Chinese Medicine) on the cognitive function and oxidative stress in mice with vascular dementia (VD) induced by cerebral ischemia/reperfusion.

METHODSThirty-two mice were randomly divided into nonnal group (n = 8), sham group (operation, but no cerebral ischemia/reperfusi6n, n = 8), model group (vascular dementia model induced by cerebral ischemia/reperfusion, n = 8), and Jiji decoction-treated group (vascular dementia model plus treatment with Jiji decoction, n = 8). Fourteen days of treatment after operation, the cognitive behavior was measured in step-through test, spatial probe test and platform test. Afterwards, to assess the levels of oxidative stress, the activity of superoxide dismutase(SOD) and content of malonaldehyde (MDA) in brain of these mice were measured.

RESULTSData from step-through test indicated that the escaping latency of Jiji decoction-treated group was prolonged and the error counts were decreased significantly ( P <0.01) compared with those of model group. Data from spatial probe test indicated that the time of entering darkroom, the time of climbing height and the time of entering bright room in Jiji decoction-treated group were shortened and the counts of climbing height were increased (P < 0.05-0.01) significantly compared with those of model group. Data from platform test showed that the escaping latency of Jiji decoction-treated group was prolonged significantly (P < 0.01) compared with that of model group. Compared with normal and sham group, the activity of SOD was decreased and the content of MDA was increased in model group significantly (P < 0.01). Compared with those of model group, the levels of SOD and MDA in Jiji decoction-treated group were improved significantly (P < 0.01).

CONCLUSIONJiji decoction could improve cognitive function of VD mice. Its mechanism might be related with the inhibition of oxidative stiess in the brain.

Animals ; Brain ; drug effects ; metabolism ; Cerebral Infarction ; physiopathology ; Cognition ; drug effects ; Dementia, Vascular ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Malondialdehyde ; metabolism ; Medicine, Chinese Traditional ; Mice ; Oxidative Stress ; drug effects ; Reperfusion Injury ; drug therapy ; Superoxide Dismutase ; metabolism

Objective To evaluate the treatment of radial head fracture of different type. Methods The clinical data of 80 radial head fracture cases from November, 1999 to December,2009 were collected and analyzed retrospectively. Among the 80 cases,12 cases received conservative treatment( all type Mason Ⅰ ) ;48 cases received open reduction internal fixation (ORIF) (4 case type Mason Ⅰ ,15 cases type Mason Ⅱ ,24 cases type Mason Ⅲ,5 cases type Mason Ⅳ ); 12 cases received removing radial head ( 4 cases type Mason Ⅲ and 8 cases type Ⅳ ).8 cases received artificial joint replacement ( 8 cases type Mason Ⅳ ). Results All cases were followed up for 1.0 -4. 1 years. According to the Mayo Elbow Performance Index, the excellent and good rate of treatment in nonoperative,ORIF,excision of the radial head as well as artificial joint replacement was 83.3 % (10/12) ,87.5 %(42/48) ,75.0% (9/12) and 100. 0% (8/8) ,respectively. Conclusions Conservative treatment can be selected for type Mason Ⅰ ;Open reduction and internal fixation can be selected for type Mason Ⅱ , type Mason Ⅲ and part of type Mason Ⅳ. The excision of radial head or mental prosthesis replacement is alternative for type Mason Ⅳ for those can not be treated with open reduction and internal fixation.

Objective To discuss the necessity of closing the peritoneum during the operation of ileal conduit after the radical cystectomy.Methods We retrospectively analyzed the clinical data of 395 patients with bladder cancer who received radical cystectomy from Jan.2014 to Sep.2016.The amount of male was 327,female was 68.The mean age was (65.8 ± 9.7) years old.Patients were divided into four groups according to the surgical method of cystectomy and urinary diversion.In group A,patients,including 78 males and 9 females,were received open radical cysectomy (ORC) with extraperitoneal ileal conduit.The mean age was (67.8 ± 9.2) years old.In the preoperative clinical staging,66 cases were less than T2 and 21 cases were more than T2.Preoperative pathological grade in 83 cases and low grade in 4 cases.In group B,patients,including 31 males and 2 females,were accepted ORC with ileal conduit without peritoneum closure.The mean age was (67.3 ± 8.7) years old.Preoperative clinical staging showed less than T2 in 25 cases,more than T2 in 8 cases,The preoperative pathological grade showed high grade in 33 cases.In group C,patients,including 112 males and 27 females,were accpeted LRC with ileal conduit without peritoneum closure.The mean age was (64.3 ± 10.5)years old.The preoperative clinical staging showed less than T2 in 107 cases and more than T2 in 32 cases.The preoperative pathological grade showed high grade in 135 cases and low grade in 4 cases.In group D,patients,including 106 males and 30 females,were accepted RARC with ileal conduit without peritoneum closure.The mean age was (65.9 ±10.0)years old.Preoperative clinical staging showed less than T2 in 103 cases and more than T2 in 33 cases.The preoperative pathological grade showed high grade in 132 cases and low grade in 4 cases.Ileal conduit without peritoneum closure means completely open the peritoneum after anastomosis of the ureter and intestine in the urinary diversion surgery without shutting down the peritoneum,which is different from the extraperitoneal ileal conduit.The operating time,blood loss,blood transfusion rate,recovery time of intestinal function and perioperative complications and rate of hydronephrosis were analyzed.Results The 395 cases completed operation successfully,no LRC or RARC had been converted to ORC.The operative time was (280.1 ± 92.3) min,(233.6 ± 99.4) min,(304.8 ± 108.9) min,(364.6 ± 86.4) min in four groups,respectively (P ＜ 0.05).The blood loss in four groups were (489.1 ± 285.6) ml,(431.8 ± 233.1) ml,(373.0 ±213.7) ml,(205.6 ± 137.8) ml,respectively (P ＜0.05).The transfusion rate in four groups were 18 (20.7％),16 (48.0％),15 (10.8％),14 (10.3％),respectively (P ＜ 0.05).The mean time to flatus in four groups were (3.7 ±1.8)d,(3.6±1.0)d,(3.5±1.2)d,(2.2±1.7)d,respectively (P ＜ 0.05).While ileal obstruction rate had no statistical difference in four groups [group A 17 cases(19.5％),group B 6 cases(18.2％),group C 27 cases(19.4％),group D 19 cases(14.0％),P =0.678].Urine leakage,intestinal leakage,lymphocyst were only occurred in group A [7 cases (8.0％),2 cases (2.3％),2 cases (2.3％)].Pyelonephritis was noticed in each group,including 14 cases(16.1％)in group A,2 cases(6.1％)in group B,9 cases (6.5％)in group C,6 cases(4.4％)in group D (P ＜ 0.05).Hydronephrosis 6 months after surgery was observed in four groups,including 15 cases(17.2％)in group A,3 cases(9.1％)in group B,7 cases(5.0％)in group C,5 cases(3.7％)in group D (P ＜ 0.05).Conclusions Ileal conduit without peritoneum closure would not increase the incidence of complications,on the contrary,it would relieve the tension of anastomosis,and reduce the occurrence of complications such as urine leakage.