Vitreoscilla Hemoglobin(VHb), with the function of increasing the growth of and product yield by a heterologous host, has been widely use in the area of fermentation, environment protection, transgenic animal and plant, recombinant protein expression, etc. Fusion protein of VHb with other enzyme or protein can enhance activity and stability of the enzyme or isolation efficiency of the protein. The reconstitution of VHb will be helpful to obtain ‘novel’ proteins which have better activity.

A novel gene,located between dnrX and drrB in the genome of daunorubicin-producing strain Streptomyces coeruleorubidus SIPI-1482,was cloned and named as dauW.The full sequence of dauW was submitted to GenBank(Accession No.EF523565).Blast result indicated that it showed high homology with dnrW in GenBank.The exact function of dauW is as yet unknown despite the possibility that it might belong to a family of FAD-dependent oxidoreductases on the basis of conserved domain analysis.dauW was cloned into expression plasmids pET-28a(+)and pET-32a(+),respectively,and was successfully expressed in E.coli DE3 after induction with IPTG.The preliminary results of the expression of dauW suggested that it might be involved in the self resistance in Streptomyces coeruleorubidus due to the increased resistance to daunorubicin in the E.coli host.

TDP-4-ketohexulose reductase, encoded by dnmV, is important in daunorubicin biosynthesis. To obtain a daunorubicin block mutant, double cross-over plasmid pYG817 was constructed by inserting apramycin resistant gene and amplified dnmV together with upstream dnmU into vector pUC18. dnmV was successfully disrupted after transformation of daunorubicin-producing strain SIPI-1482 by pYG817. Daunorubicin was absent from metabolites of the resulting transformant, and its biosynthesis can be reconstituted by introducing dnmV expression plasmid into the disruptant, although the yield is lower than wild-type SIPI-1482, according to HPLC analysis. This mutant can be a good candidate for production of anthracycline such as epi-daunorubicin by introducing exogenous gene into the host.

TTI gene coding for Tsetse thrombin inhibitor was modified with E.coli bias codon and expressed in Escherichia coli with high efficiency.Recombinant protein was purified to more than 98% purity.Assay for enzyme activity determination was set up.The result showed that the fusion protein exhibited inhibiting activity for thrombin.Inhibitory rate of purified TTI was 73% when concentration of thrombin and substrate was 10U/ml and 250?mol/L respectively.Inhibition pattern was determined as competitive with Ki at 35?mol/L.

Objective To evaluate the short term effect of bipolar arthroplasties in aged cases with intertrochanteric fractures.Methods There were 23 males and 15 females(at age of 70-93 years, average 76 years)with unstable intertrochanteric fractures treated with the third generation cementing techniques and bipolar arthroplasties that were followed up for average 2.4 years.The clinical effect was evaluate with Harris scale,X-ray films and complications.Results Of all,34 cases(89.5%)could walk freely.The average Harris scale was 84.2 points.The average period until walk with full-weight load was 5.6 weeks.The greater trochanters was united in 35 cases(92%)during average 4.2 months.No peri-prothetic ossteolysis and loosening or subsidence occurred.Conclusions Cemented bipolar arthro- plasty has good advantages of reduced laying up period and few complications.The short-term outcome is satisfactory hut the long-term outcome needs deeper observation.

To improve the growth enhancement activity of Vitreoscilla hemoglobin(VHb), Vitreoscilla hemoglobin gene(vgb) was mutated by error-prone PCR and then reconstituted by DNA shuffling. The shuffling library was constructed by inserting the shuffled genes into the downstream of vgb natural promoter and transforming them into E.coli DH5?. Mutated active VHb proteins were first screened in test tubes according to host cell pellets color and then in shake flasks according to host pellets wet weight .One active mutant protein, VHb′042506, was obtained after second screening. It could increased the host wet weight by 31.25% and 58.75% than that of the control which bearing natural VHb under microaerobic and extremely microaerobic conditions, respectively. Sequencing and alignment results showed that 11 nucleotides were mutated, thus resulted in 4 amino acids changes occurred in this mutant protein. CO difference spectrum test also indicated that it had higher specific absorption.

The object of this study is to preparate the berberine hydrochloride (BBH) resin compound with taste masking effect. We took the BBH as the model drug and Amberlite IRP69 as the drug carriers, uncovered the curve of solubility of BBH in different cosolvent with a certain range of temperature, and then used it to calculate the parameters during the preparation of the complex such as adding quantity of BBH and the reaction temperature. Afterwards, the characteristic and in vitro release experiments were studied to verify the formation and predict the in vivo release behavior of the complex. The results showed that in the condition of using 60% ethanol as a cosolvent and stirring at 50 ℃ for 1 h, the drug loading and drug availability of the complex are at about 35% and 64%, respectively, and has a better taste-masking effect. In this study, a method was provided for preparing a taste-masking preparation of BBH.

Cannula ; Catheters ; Drainage ; Duodenal Diseases ; Humans ; Intestinal Fistula/surgery*

OBJECTIVETo identify the potential determinants of return to work (RTW) following work-related injury.

METHODSA historical cohort of workers with occupational injury in a state-owned locomotive vehicle company in central China was followed up for RTW. Demographic, employment and medical information was retrieved from the company archival documents; and post-injury information was interviewed by questionnaires. Univariate analysis and Cox Regression Model were used to examine the associations between potential determinants and outcomes of RTW.

RESULTSThree hundred of the 323 included cases (92.9%) eventually returned to work after the median absence of 43 days (average of 49.2 days). Factors from socio-demographic, clinical, economic and psychological domains were found affecting RTW in the univariate analyses. The multivariate analysis indicated that age, injury severity, injury locus, injury nature, pain in the injury locus, self-reported health status and pre-injury monthly salary were significant determinants of RTW.

CONCLUSIONSThere are multidimensional factors affecting RTW after occupational injury. Proper clinical treatment and rehabilitation, as well as economic and social support to facilitate workers' RTW would be the priorities for intervention. Future studies should be conducted in a larger representative sample to confirm the findings and to develop a multidisciplinary intervention strategy towards promoting RTW.

Adult ; Cohort Studies ; Female ; Humans ; Male ; Middle Aged ; Occupational Health ; statistics & numerical data ; Occupational Injuries ; Proportional Hazards Models ; Retrospective Studies ; Sick Leave ; Surveys and Questionnaires ; Work

?-lactamase inhibitor research is popular for its potential on ?-lactam antibiotics resistant strain.A ?-lactamase binding peptide SIPIS04-01 was obtained by the yeast two-hybid system.In vitro assay showed that it can inhibit the ?-lactamase activity.In order to improve the expression level of the recombinant peptide,a two-copy expression plasmid pYG563 was constructed by random orientation tandem repeat method after codon modification,the two-copy plasmid was successfully expressed and the product was increased by 48.4% than that of one-copy plasmid.Purified peptide showed inhibitory activity against TEM-1 ?-lactamase in vitro and the inhhibitory constant Ki was measured.