Trehalose, a compatible solute, is widely used in food, cosmetics, pharmaceutical products and organ transplantation. Nowadays, trehalose is mostly produced by enzymatic synthesis with many secondary products and lowpurity. In this study, high amount of trehalose was produced by recombinant E. ccli fermentation. First, a bifunctional trehalose gene TPSP was amplified from genome of C. hutchinscoii. Second, an expression vector pTac-HisA containing TPSP was constructed and transformed into the host E. coli. Expression of this bifunctional enzyme-TPSP converted glucose to trehalose. The result suggested that TPSP from C. hutchinsonji has been successfully expressed in E. ccoi. High amount of extracellular trehalose generated from glucose by whole-cell catalysis and After optimization, the production of trehalose in shake flasks was improved to 1.2 g/L and the relative conversion rate reached 21%. The production in bioreactor reached 13.3 g/L and the relative conversion rate reached 48.6%. It is the first time to realize the functional expression of the bifunctional enzyme-TPSP of C. hutchinsonii in E. coli and achieved the conversion form glucose to trehalose. This study laid a foundation for industrial large-scale production of trehalose.
Bioreactors ; Catalysis ; Escherichia coli ; genetics ; Glucose ; Glucosyltransferases ; Industrial Microbiology ; Organisms, Genetically Modified ; Trehalose ; biosynthesis

Objective To investigate the testing capabilities of clinical transfusion laboratories in medical institutions in Beijing for the regulatory authorities to formulate administrative policies in this regard.Methods Experts assigned by Beijing Clinical Transfusion Quality Control Center made on-site inspections at the transfusion laboratories in medical institutions using quality control products.They recorded the complete testing process of the operators as well as the instruments,detection reagents in use and the testing results,with statistics and analysis made to the data so collected.Results The pass rate of these on-site inspections was lower than that of the external quality assessment.Some laboratories failed to complete the testing of the quality control products in time and the actual operations in some laboratories were inconsistent to the guidelines.55.9％ of level Ⅰ hospitals and 25.6％ of level Ⅱ hospitals were found with insufficient and inadequate instruments and process layout to meet the needs of clinical blood transfusion.Some of the technicians were found without sufficient trainings in their professional knowledge and basic skills,resulting in their poor competence against emergency cases and weakness in independent problem solving.In addition,the records of detection process and results were found to be substandard.Conclusions Transfusion laboratories in Beijing need to improve their testing capabilities in general.

A total of 58 patients with simple septic shock were recruited from intensive care unit and divided into control group (n =28) and treatment group (n =30) according to treatment modalities.The control group was routinely treated.The treatment group received continuous veno-venous hemofiltration (CVVH) for 10-14 days plus routine measures.After 3 days,the score of acute physiology and chronic health evaluation-Ⅱ (APACHE-Ⅱ) of the treatment group was better than that of the control group (P ＜ 0.01).And sequential organ failure assessment (SOFA),left ventricular end diastolic diameter (LVEDD),left ventricular end systolic diameter (LVEDS) and left ventricular ejection fraction (LVEF) improved significantly (P ＜ 0.05).And the relevant blood biochemical parameters improved significantly better than the control group (P ＜ 0.05).In the group CVVH,there were mortality (n =7,23％) and multiple organ failure (MODS) (n =6,20％) ; In the control group,mortality (n =14,50％) and MODS (n =13,46％).The mortality rate had inter-group differences of statistical significance (x2 =4.38,P ＜0.05).Thus early volume resuscitation plus CVVH had excellent curative efficacies for septic shock.

Objective This study explored the expression of polyclonal protein SUZ12 in patients with intrahepatic cholangiocarcinoma(ICC),and its role in predicting the survival and treatment of ICC patients.Methods The expression of SUZ12 and p16INK4a was detected by immunohistochemical assay in 207 liver tissue samples including ICC patients,BilIN-1,-2,-3 and non-tumor-like cholangiocarcinoma.The expression of these proteins was assessed to be related to the pathological characteristics of the ICC patients receiving chemotherapy and the outcome of survival as well as the subsequent chemotherapy response.Results The expression level of SUZ12 was gradually increased from non-neoplastic bile duct tissue to BilIN-1,-2,-3 and ICC.The expression of p16INK4a protein was expressed in non-neoplastic-like cholangiocarcinoma,but it decreased gradually in BilIN-1,-2,-3 and ICC tissues.SUZ12 expression was associated with undifferentiated ICC,lymph node metastasis and advanced cancer.Kaplan-Meier curve analysis showed that ICC patients with high expression of SUZ12 had a significant reduction in overall survival and disease-free survival in comparison with ICC patients with the low expression of SUZ12.SUZ12 expression was significantly associated with overall survival of patients receiving adjuvant gemcitabine-based chemotherapy(AGC).Conclusion SUZ12 expression is able to predict the overall survival and disease-free survival of ICC patients with adjuvant gemcitabine-based chemotherapy.

Objective To investigate the effect of depression symptoms on surgical outcome of posterior decompression among cervical spondylotic myelopathy (CSM) patients. Methods Between October 2006 and October 2011 in our hospital, lami?nectomy or laminoplasty was performed in 396 cases that were enrolled in the study. There were 132 males and 264 females with an average age of 60.2 years(ranged,39-84 years). All patients were divided into depression group and non?depressed group by the 21?item Beck Depression Inventory (BDI). There were no statistically significant differences between groups in age, sex, smok?ing status, duration of symptoms, and employment status (whether in the current working). The Japanese Orthopedic Association (JOA) scores, Neck disability index (NDI) and visual analogue scale (VAS) were compared after 1.5 months postoperatively. Re?sults All of 396 cases were followed up. The mean follow up duration was 32 months (range,24-50 months). There were no statistically significant differences in the CCI decline (7.1%±2.1% versus 6.8%±1.5%), expansion degree[(130.9±7.0) mm2 versus (150.8 ± 5.2) mm2] and the drift?back distance of the spinal cord [(5.7 ± 1.2) mm versus (6.2 ± 0.8) mm]. However, pa?tients with continuous depression showed poorer improvement than non?depressed patients in the surgery outcome: JOA im?proved (1.42±0.56 versus 6.76±3.12); NDI declined (7.31±2.18 versus 21.11±11.36); and VAS lightened (16.08±19.76 versus 23.85±20.79). Conclusion Depression contributed a dissatisfied surgery outcome after posterior decompression on functional re?covery, disability index and pain scores among patients of the cervical spondylotic myelopathy.

OBJECTIVETo study preliminarily on the relationship between properties of the mixture composed of Tongsaimai extract and different excipients and pellet molding.

METHODThe multivariate regression analysis was used to investigate the correlation of different mixture and pellet molding by measuring the cohesion, liquid-plastic limit of mixture, and the powder properties of pellets. The weighted coefficients of the powder properties were determined by analytic hierarchy process combined with criteria importance through intercriteria correlation.

RESULTThe results showed that liquid-plastic limit seemed to be a major factor, which had positive correlation with pellet molding, while cohesion had negative correlation with pellet molding in the measured range.

CONCLUSIONThe physical properties of the mixture has marked influence on pellet molding.

Dosage Forms ; Drug Compounding ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Excipients ; chemistry ; Particle Size ; Powders ; Surface Properties

Group II chaperonins, which assemble as double-ring complexes, assist in the refolding of nascent peptides or denatured proteins in an ATP-dependent manner. The molecular mechanism of group II chaperonin assembly and thermal stability is yet to be elucidated. Here, we selected the group II chaperonins (cpn-α and cpn-β), also called thermosomes, from Acidianus tengchongensis and investigated their assembly and thermal stability. We found that the binding of ATP or its analogs contributed to the successful assembly of thermosomes and enhanced their thermal stabilities. Cpn-β is more thermally stable than cpn-α, while the thermal stability of the hetero thermosome cpn-αβ is intermediate. Cryo-electron microscopy reconstructions of cpn-α and cpn-β revealed the interwoven densities of their non-conserved flexible N/C-termini around the equatorial planes. The deletion or swapping of their termini and pH-dependent thermal stability assays revealed the key role of the termini electrostatic interactions in the assembly and thermal stability of the thermosomes.
Acidianus ; metabolism ; Adenosine Triphosphate ; metabolism ; Amino Acid Sequence ; Cryoelectron Microscopy ; Hydrogen-Ion Concentration ; Molecular Sequence Data ; Mutation ; Nucleotides ; metabolism ; Protein Binding ; Protein Folding ; Protein Stability ; Protein Structure, Quaternary ; Sequence Alignment ; Static Electricity ; Temperature ; Thermosomes ; chemistry ; genetics ; metabolism

ObjectiveTo investigate the demand, supply and satisfaction of the Chinese people on assistive technology. MethodsBased on the data from the World Health Organization rapid Assistive Technology Assessment (rATA) questionnaire in six provinces and cities in China, the assistive technology of the whole population in China was described from three aspects of demand, supply and satisfaction. ResultsA total of 15 234 valid questionnaires were obtained. In terms of demand, the proportion of visual impairment was the highest in all the functional impairment categories, the highest demands in the list were glasses (low-vision, short/long distance etc.) and walking sticks (canes/sticks, tripod and quadripod), while the highest demand outside the list was anti-skid pad. In terms of supply, the private sector was the main one, followed by government facility and public hospital, and most users took "paid out-of-pocket (self)" to obtain assistive technology, the distance to access the assistive technology was concentrated at "less than five kilometers", and the biggest obstacle to obtain assistive technology was lack of knowledge about assistive technology. Overall satisfaction with assistive technology was optimistic, the adaptation degree and service quality of assistive technology were the most unsatisfactory factors for users. ConclusionIn China, the satisfaction of assistive technology is optimistic, and there is a few amount of unmet assistive technology. It is necessary to propagandize assistive technology, and its manual should be readable, the assistive technical service team should be multidisciplinary, enterprises should promote independent innovation, and the professionals should be regulated.