Microbial lipases have been widely used in traditional industries,as well as in emerging biocatalysed areas owing to their ability to catalyze a variety of reactions in aqueous and non-aqueous media.Therefore,it is very important to enhance amount of lipase production by recombinant overexpression for meeting market demand.A critical review of main and novel strategies which have been employed for recombinant expression of microbial lipases are presented,including codon optimization,fusion and co-expression,dual expression system based on hybrid promoters,homologous overexpression,cell surface displaying and high-throughput screening based on gene library of expression.These new technologies are gradually coming to the forefront in the recombinant expression of lipase,especially for cell surface displaying and high-throughput screening based on gene library of expression.Meanwhile,several recombinant expressions for representative microbial lipases were also introduced and discussed,which are available for consultation when attempting to overexpress any lipases by scientists and industrialists.

Small and medium-sized enterprises (SMEs) are important components in Chinese medicine industry. However, the lack of big brand is becoming an urgent problem which is critical to the survival of SMEs. This article discusses the concept and traits of Chinese medicine of big brand, from clinical, scientific and market value three aspects. Guided by market value, highlighting clinical value, aiming at the scientific value improvement of big brand cultivation, we put forward the key points in cultivation, aiming at obtaining branded Chinese medicine with widely recognized efficacy, good quality control system and mechanism well explained and meanwhile which can bring innovation improvement to theory of Chinese medicine. According to the characters of SMEs, we hold a view that to build multidisciplinary research union could be considered as basic path, and then, from top-level design, skill upgrading and application three stages to probe the implementation strategy.
China ; Drug Industry ; economics ; standards ; trends ; Drugs, Chinese Herbal ; economics ; standards ; Medicine, Chinese Traditional ; economics ; standards ; trends ; Plants, Medicinal ; growth & development ; Quality Control

Purpose:To study the effect of inhibition of telomerase activity and cell cycle by transcriptase telomerase inhibitors (3′ azido 3′ deoxythymidine, AZT) on squamous cell carcinoma of tongue in vitro.Methods:Human squmous cell carcinoma of tongue cell line Tca8113 was used as target cell. Telomerase activity was determined by TRAP PCR ELISA in untreated and treated Tca8113 by AZT, cell cycle phases were analyzed by flow cytometry. Results:Telomerase activity of Tca8113 was significantly inhibited when treated with AZT, and the effect of inhibition was dose dependant (rate of telomerase activity treated with AZT in 0.3, 0.6, 1.0, 1.5mol 10 -1 was 0.69 0.03, 0.61 0.08, 0.53 0.11, 0.50 0.02 respectively, rate of telomerase activity treated without AZT was 0.76 0.06). Cell cycle of treated Tca8113 was changed with marked increase in G 2 /M phase compared with untreated Tca8113 (62.8% vs 19.7%, P

The fermentation conditions of lipase production by Geotrichum candidum Y162 were optimized. Initially, the most suitable carbon olive oil, nitrogen source soybean flour and NH4Cl, salt BaCl2 and MgCl2 were selected according to single factorial experiments respectively. Based on the result, screening methodology Plackett-Burman design was used to evaluate the effects of twelve factors related to lipase production and three statistically significant factors olive oil, BaCl2 and NH4Cl were selected. The path of steepest ascent was used to approach the optimal region of lipase production subsequently. Then, the optimal combined concentration for maximum enzyme activity were further optimized by response surface methodology and determined as follows: olive oil 2.35%, BaCl2 0.36%,and NH4Cl 4.69%.The optimization of culture conditions of G.candidum Y162 led to a 2.25-fold increase in lipase production relative to initial result 14.16 U/ml, which indicate that single factor in combination with response surface methodology is an effective method for optimization of lipase production conditions by G.candidum Y162.

?AIM:To evaluate the clinical outcomes of Nd:YAG laser capsulotomy in the treatment of early stage capsular block syndrome ( CBS) .?METHODS:Eighteen patients (21 eyes) with early stage capsular block syndrome were treated using Nd:YAG laser by anterior capsulotomy only or combined with posterior capsulotomy from January 2010 to July 2015 in Anyang Eye Hospital. Uncorrected distance visual acuity, intraocular pressure, spherical equivalent, depth of anterior chamber were observed preoperatively and 2wk postoperatively.?RESULTS:Seventeen eyes simply underwent peripheral anterior capsulotomy with Nd:YAG laser. Four eyes were combined with posterior capsulotomy. Compared with preoperative, uncorrected distance visual acuity improved, intraocular pressure returned to normal, degree of myopia reduced, depth of anterior chamber had deepened.? CONCLUSION: Nd: YAG laser capsulotomy is an effective treatment for early stage capsular block syndrome.

A UPLC method has been developed in the current investigation for simultaneous determination of nine chemical markers of Bletilla striata, 4-hydroxymethylphenyl beta-D-glucoside, blestroside, dactylorhin A, militarine, dihydrophenanthrene 5, gymnoside V, dihydrophenanthrene 1, benzylphenanthrene 3 and gymnosides IX. Separation was performed at 45 degrees C on an ACQUITY UPLC BEH C18 column (2.1 mm x 150 mm, 1.7 microm) with a gradient solvent system of acetonitrile-water as the mobile phase. The flow rate was 0.3 mL x min(-1), the detection wavelength was 280 nm. The results showed that the nine chemical markers could be well resolved and that in the selected linear range, all calibration curves of the nine chemical markers showed good linearity (r > or = 0.999 3). The recoveries (n = 6) were in the range of 98.15% - 102.2% and RSDs were between 2.1% - 3.6%. The data suggested that the developed UPLC-UV method had good reproducibility, robustness, and accuracy, which was suitable for the quality control of Bletilla striata. Applications of the method showed that the nine chemical markers had higher contents in the wild B. striata than in the cultivated ones.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Magnoliopsida ; chemistry ; Rhizome ; chemistry

OBJECTIVEExpress and purify four single-stranded DNA-binding (SSB) proteins, and evaluate the binding of SSB proteins on HCV RNA.

METHODSThe expression plasmids of four SSB proteins were conducted, termed TTH, SSOB, KOD and BL21, respectively. The BL21 (DE3) was transformed by the expression plasmid of TTH, Transetta (DE3) were transformed by the expression plasmid of SSOB, KOD and BL21, then protein expression was induced with IPTG, the expression products were analysised by SDS-PAGE. To evaluate the binding of SSB on HCV RNA, RNA-SSB protein complexes were applied to a 1.2% TAE agarose gel.

RESULTSSuitable competent cells were transformed with the expression plasmids, induced by IPTG. SSB proteins were purified by affinity chromatography, to visualize their purity all SSB proteins were applied to SDS-PAGE analysis. All four proteins showed single clear bands. We have successfully obtained the SSB protein expression plasmid, expressed and purified SSB protein. TAE agarose gel electrophoresis was used to confirm SSB protein-RNA binding activity. The each of SSB-RNA complex migrated more slowly than the sole RNA, which suggested SSB protein could specifically bind to RNA.

CONCLUSIONSWe have expressed and purified four SSB proteins, and for the first time found that SSB protein can bind HCV RNA. Our results may provide a basis for future studies of the novel functions of SSB proteins on RNA.

DNA, Single-Stranded ; genetics ; metabolism ; DNA-Binding Proteins ; chemistry ; isolation & purification ; metabolism ; Hepacivirus ; Hepatitis C ; metabolism ; virology ; Humans ; Molecular Weight ; Protein Binding ; RNA, Viral ; genetics ; metabolism

Adult ; Efficiency ; Female ; Humans ; Male ; Middle Aged ; Psychometrics ; Surveys and Questionnaires

Child ; Child, Preschool ; Craniocerebral Trauma ; etiology ; surgery ; Female ; Humans ; Male

0.05)respectively,when the whole radiotherapy was finished,the CR was 53.3% and 25.8%(P