A 1.8 kb HinfI fragment carrying the chitinase gene(chiA) from Serratia marcescens was isolated from the plasmid pLCHIA and was inserted into the SmaI site downstream of the strong Ptac promoter in the expression vector pKK223-3,yielding the plasmid pKChiA.The 2.1 kb PtacChiA fusion fragment was excised by BamHI from the plasmid pKChiA and was inserted into the single BamHI site present within the plasmid pMC71A,thus generating the plasmid pMChiA.High levels of chitinase were produced by the E.coli strains HB101 and JM105 carrying the plasmid pKChiA or the plasmid pMChiA,with the amount of production were higher 1～3 fold than that produced by the strains carrying the plasmid pLCHIA.

Air Pollutants, Occupational ; analysis ; Solvents ; analysis

Objective To investigate the effects of total anthraquinone in rheum on aquaporin 2 and aquaporin 4 expression in rat kidney and explore its diuresis mechanism.Methods Thirty-two SD rats were randomly divided into control group,low dose group,medium dose group and high dose group.Total anthraquinone in rheum was administered to rats at different doses.Urinary volume of 24 h,Na+ concentration and osmolality were detected.Rats were sacrificed 5 days later.Blood samples were taken from the abdominal aorta to detect blood biochemical indicators. Kidneys of rats were removed to detect AQP2, AQP4 expression through immunohistochemistry,Western blot,and RT-PCR. Results Compared with control group,there were significantly increased 24 h urine output of rats in medium and high dose group[(16.21±1.96),(18.16±1.8) ml vs(13.85±1.25)ml,P＜0.05].24 h urine output in low-dose group did not change significantly.AQP2 protein and mRNA expression were significantly decreased in rats'kidneys of medium and high dose group (P＜0.01),The AQP4 protein and mRNA expression was significantly down-regulated in high dose group (P＜0.01).In medium does group,the AQP4 protein expression was down-regulated (P＜0.01),without significant decrease in the mRNA expression.Protein and mRNA expression of AQP2 and AQP4 did not significantly change in low dose group.Conclusion Total anthraquinone in rheum can reduce the expression level of AQP2 and AQP4 in rat kidney,which is probably one mechanism of diuresis caused by rheum.

Traditional herbal medicines, Panax ginseng, Panax quinquefolium and Panax notoginseng, attract our attention for their extensive and powerful pharmacological activities. Ginsenosides are the main active constituents of these medicinal herbs. The related glycosyltransferases involved in ginsenoside biosynthesis are the key enzymes which catalyze the last important step. Modification of ginsenoside aglycones by glycosyltransferases produces the complexity and diversity of ginsenosides, which have more extensive pharmacological activity. At present, ginsenoside aglycones and compound K have been obtained by synthetic biology. As the last step of ginsenoside biosynthesis, glycosylation of ginsenoside aglycones has been studied intensively in recent years. This review summarizes the basic strategies and research advances in studies on glycosyltransferases involved in ginsenoside biosynthesis, which is expected to lay the theoretical foundation for the in-depth research of biosynthetic pathway of ginsenosides and their production by synthetic biology.
Biosynthetic Pathways ; Ginsenosides ; biosynthesis ; Glycosyltransferases ; metabolism ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Synthetic Biology

According to morphological and microscopic characteristics, a high-efficient decolorizing fungus, the strain Asaw117, was identified as Aspergillus awamori. Selecting eight different dyes from Azo dyes, anthraquinones dyes and oxygen Quinones dyes, the decolorizing assays of various dyes showed that the strain Asaw 117 was the highest decolorizing potential to 0.1 g/L Vat Blue RSN, the discoloration rate up to 100 percent. Comparing to different kinds of medium and several of carbon and nitrogen sources, the strain had the best decolorizing efficient although grew slower in the Czapek medium, otherwise, grew quicker and decolorizing efficient lower in the PDF medium. It could use Vat Blue RSN as a nitrogen source, but not as a carbon source. The medium composing of saccharose and ammonium nitrate as carbon and nitrogen sources was decolorizing potential markedly during different combinations of carbon and nitrogen sources. So the strain has good potential for the dyeing wastewater treatment

For the establishment of chemical library of protoberberines provided for the bio-activity screening, the target compounds were synthesized by thermal degradation and nucleophilic substitution reactions with the bio-active alkaloid, palmatine (1), as the raw material, and their structures were identified and conformed by 1H-NMR and MS spectra. Among them, 13 compounds were new.
Berberine Alkaloids ; chemical synthesis ; chemistry ; Drugs, Chinese Herbal ; chemical synthesis ; chemistry ; Molecular Structure

OBJECTIVETo study the relationship of TCM syndrome type of gastric mucosal epithelial growth factor (EGF), vascular endothelial growth factor (VEGF) and proliferative cell nuclear antigen (PCNA) in patients with chronic atrophic gastritis (CAG) for exploring the essence of TCM type and providing a theoretical basis of clinical treatment.

METHODSTCM syndrome type of 200 patients with diagnosis of CAG confirmed by fibro-gastroscope and pathological examination were differentially classified, and the expressions of EGF, VEGF and PCNA in different types were determined using immunohistochemistry.

RESULTSPatients were differentiated as Pi-Wei deficiency type (Type I ) in 72; Gan-Wei disharmony type (Type II ) in 43; Pi-deficiency with qi stagnation type (Type III) in 32; Wei-yin deficiency type (Type IV) in 24; Pi-Wei damp-heat type (Type V) in 14; and Wei-collateral stasis obstruction type (Type VI) in 5. The difference of PCNA expression level between Type II with Type I , III and IV was significant (P < 0.05). No significant difference in expression levels of EGF and VEGF was found among the 6 types (P > 0.05).

CONCLUSIONType I and II were the dominant TCM syndrome types in CAG patients; the high expression of PCNA might be a diagnostic evidence for Gan-Wei disharmony syndrome.

Adult ; Aged ; Aged, 80 and over ; Diagnosis, Differential ; Endothelial Growth Factors ; biosynthesis ; Female ; Gastric Mucosa ; metabolism ; pathology ; Gastritis, Atrophic ; diagnosis ; metabolism ; Humans ; Immunohistochemistry ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Syndrome ; Vascular Endothelial Growth Factor A ; biosynthesis

OBJECTIVETo study the therapeutic effects of posterolateral depression fractures of the tibial plateau through a modified anterolateral approach.

METHODSFrom February 2011 to January 2012,13 patients with posterolateral depression fractures of the tibial plateau were treated through a modified anterolateral approach. There were 8 males and 5 females, ranging in age from 28 to 59 years old (49.2 years old on average). Data from patients were collected retrospectively as follows: X-ray, time of fracture healing and the complications of fracture healing. The patients were evaluated both clinically and radiologically according to the Rasmussen score system.

RESULTSAll the patients were followed up, and the duration ranged from 6 to 18 months (mean 13.7 months). All the patients got bony union. The average radiographic bony union time was 15.1 weeks (ranged, 11 to 17 weeks). No case of secondary articular depression was found. No complications such as malunion or joint stiffness were found. But 1 patient had superficial infection and 1 patient had common peroneal nerve injury. According to the Rasmussen score system,the mean radiological score was 16.50 ± 0.67 (ranged, 13 to 18), and the mean functional score was 25.20 ± 2.21 (ranged, 13 to 30). The mean range of knee motion was (125.3 ± 9.3)° (ranged, 0° to 135°).

CONCLUSIONTreatment of depression fractures of posterolateral tibial plateau with a modified anterolateral approach is a safe method with effective exposure, due to its stable fixation and relatively good outcome with minimal soft-tissue complications. It is regarded as an ideal procedure for depression fractures of posterolateral tibial plateau.

Adult ; Female ; Fracture Fixation, Internal ; methods ; Fracture Healing ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Tibial Fractures ; surgery

Aged ; Brain Abscess ; etiology ; Brain Neoplasms ; complications ; secondary ; Carcinoma, Squamous Cell ; complications ; secondary ; Humans ; Male ; Neoplasms, Unknown Primary ; Parietal Lobe

Pichia pastoris is one of the most important systems used in the field of molecular biology for the expression of recombinant proteins. The system has advantages of high expression, high stability, high secretion, easy high-density fermentation and low cost. Many factors affect the expression of recombinant protein, such as gene copy number, codon usage preference, type of promoter, molecular chaperones, glycosylation, signal peptide and fermentation process. In this review, research advances of the above aspects are summarized, which lay a foundation for improving the expression of recombinant proteins in P. pastoris.
Fermentation ; Gene Dosage ; Glycosylation ; Molecular Chaperones ; Pichia ; metabolism ; Promoter Regions, Genetic ; Protein Sorting Signals ; Recombinant Proteins ; biosynthesis