OBJECTIVE:To prepare risperidone injectable biodegradable implants and study its in vitro release. METHODS: Polylactide-co-glycolide copolymer(PLGA)was chosen for carrier and N-Methyl pyrolidone(NMP)as solvent to prepare risperidone injectable biodegradable implants. The percentage of accumulative drug release within 30 days,the change in molecular weight of copolymer and superficial opening diameters of copolymer in function time were observed. RESULTS:Drug release curve of risperidone injectable biodegradable implants for 30 days was stable with small burst. The percentage of accumulative drug release was 89.01% and the percentage of burst accounted for 13.8% at the first 24 hours. The average molecular weight of copolymer decreased to about 10 000 from 43 000 after 30 days and the size of the superficial openings became larger and larger. CONCLUSION:Risperidone injectable biodegradable implants can release continuously and stably in vitro for 30 days.

Objective To investigate the distribution of pathogenic bacteria,drug sensitivity,and the relationship between drug sensitivity and Escherichia coli(E.coli) induced enzymes in childhood diarrhea in the last 2 years in Chongqing area,so as to provide important evidence for pediatric clinical therapy.Methods Thirty-one E.coli induced enzymes,extended spectrum ?-laetamases(ESBLs),cephalosporinase(AmpC)detected in different phenotype methods,and drug sensitivity was measured in paper strip method,and the specimens were collected from children′s hospital affiliated to chongqing university of medical sciences from Jan.2005 to Dec.2006 were determined.Among the total,there were 18 enteropathogeic E.coli(EPEC) strains,8 enterotoxigenic E.coli(ETEC) strains and 5 enteroinvasive E.coli(EIEC) strains.In addition,drug resistance tests by paper strip included chloramphenicol(CHL),amikacin(AMK),gentamicin(GEN),norfloxacin(NOF),ciproflocacin(CIP),cefazolin(CEZ),cefoperazone(CPZ),ceftriaxone(CRO),ceftazidime(CAZ),cefotacime(CTX),cefepime(FEP),imipenem(IPM).SPSS 12.0 software was used to analyze the data.Results Three point two percent of the 31 E.coli were drug resistant to IPM,and 35.5%,38.7% to NOF,CIP individually,but more than 60% to AMK,GEN,even more than 67.7% towards cephalosporin(except ceftazidime and cefepime);the gross enzyme-produced rate was 87.1%,rate of single ESBLs,AmpC,and induction of both enzymes simultaneously presented 64.5%,6.5%,16.1% respectively;and there was marked difference in drug resistance when bacteria that produced single AmpC versus bacteria that produced single ESBLs or that produced both ESBLs and AmpC(Pa﹤0.05).Conclusions The relationships among enzyme′s quantity,sort and bacterial resistance are different.These data show E.coli infected by bacterial diarrhea children in Chongqing due to a high rate of induced enzymes,and their drug resistance vary according to the state of induced enzymes.

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

Bacteremia ; etiology ; Female ; Humans ; Infant, Newborn ; Listeria monocytogenes ; isolation & purification ; Male

A RP-HPLC method was developed to evaluate the quality of Picrasmae Ramulus et Folium by simultaneous determination of five constituents including 1-hydroxymethyl-beta-carboline (1), 1-methoxicabony-beta-carboline (2), 4-methoxy-5-hydroxy-canthin-6-one (3), 4, 5-dimethoxy-canthin-6-one (4) and maackiain (5) in Picrasmae Ramulus et Folium. The samples were separated on a Kromasil RP-C18 (4.6 mm x 250 mm, 5 microm) column eluted with acetonitrile and 0.1% phosphoric acid as mobile phases in gradient mode. The detection wavelength was set at 254 nm. The calibration curves and linearity of the above five standards were determined as (1) Y = 6 525.6X + 37.25 (0.009-1.780 microg, r = 0.996 8), (2) Y = 3 662.3X + 41.55 (0.005-0.920 microg, r = 0.999 5), (3) Y = 3763.1X + 146.87 (0.015-3.060 microg, r = 0.999 0), (4) Y = 2 174.1X + 21.52 (0.003-0.620 microg, r = 0.999 5), and (5) Y = 276.25X + 7.65 (0.010-1.960 microg, r = 0.998 9), respectively. The method is simple and repeatable, and can be used for the quality assessment of Picrasmae Ramulus et Folium.
Alkaloids ; analysis ; Calibration ; Carbolines ; analysis ; Chromatography, High Pressure Liquid ; methods ; Chromatography, Reverse-Phase ; methods ; Flavonoids ; analysis ; Indole Alkaloids ; analysis ; Picrasma ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Pterocarpans ; analysis ; Reproducibility of Results

OBJECTIVETo explore the effect of qidong huoxue decoction (QHD) on inflammatory factors and Toll-like receptor (TLR4) mRNA expressions in acute lung injury (ALI) rats.

METHODSTotally 50 healthy male SD rats were randomly divided into the blank control group, the lipopolysaccharide (LPS) model group, low, middle, high dose QHD groups according to body weight, 10 rats in each group. Rats in low, middle, high dose QHD groups were intragastrically administered with QHD at 4, 8, and 16 mL/kg 24, 12 h before modeling and 12 h after modeling, respectively. Normal saline was intragastrically administered to rats in the blank control group and the LPS model group. An ALI rat model was established using intratracheal instillation of LPS. Rats were killed after 24-h modeling. Then the bronchoalveolar lavage fluid was prepared. Contents of TNF-α, IL-1β, and L-10 were detected using ELISA. TLR4 mRNA expressions were determined byreal time PCR.

RESULTSCompared with the blank control group, contents of TNF-α, IL-1β , and IL-10 increased (P <0. 01), TLR4 mRNA expressions also increased in the LPS model group (all P <0. 01). Compared with the LPS model group, contents of TNF-α and IL-1β decreased (P <0. 05, P <0. 01), IL-10 levels increased (P <0. 01) , TLR4 mRNA expressions were also reduced (P <0. 01), in high and middle dose QHD groups. Compared with the high dose QHD group, con- tents of TNF-α and IL-1β increased in middle and low dose QHD groups (P <0. 05); IL-10 levels decreased (P <0. 05) in the low dose QHD group(P <0. 05), TLR4 mRNA expressions also increased in the low dose QHD group (P <0. 05). Compared with the middle dose QHD group, IL-10 levels was reduced, but TLR4 mRNA expressions increased in the low dose QHD group (P <0. 05).

CONCLUSIONSQHD had the protective effect on LPS induced ALI rats. Its mechanism might be associated with inhibiting TLR4 mRNA expressions, leading to decreased pro-inflammatory cytokines such as TNF-α and IL-β, elevated anti-inflammatory cytokine IL-10, and thereby, correcting unbalanced inflammation.

Acute Lung Injury ; genetics ; metabolism ; Animals ; Anti-Inflammatory Agents ; Bronchoalveolar Lavage Fluid ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; Interleukin-10 ; metabolism ; Interleukin-1beta ; metabolism ; Lipopolysaccharides ; Male ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

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

Lanosterol synthase is encoded by the erg7 gene and catalyzes the cyclization of 2, 3-oxidosqualene, which is a rate-limiting step of the inherent mevalonate (MVA)/ergosterol metabolic pathway in Saccharomyces cerevisiae. The intermediate 2, 3-oxidosqualene is also the precursor of triterpenoids. Therefore, the cyclization of 2, 3-oxidosqualene is the key branch point of ergosterol and triterpenoids biosynthesis. Down-regulation of 2, 3-oxidosqualene metabolic flux to ergosterol in S. cerevisiae may redirect the metabolic flux toward the triterpenoid synthetic pathway reconstructed by the synthetic biology approach. To construct erg7 knockout cassette harboring the loxP-Marker-loxP element, long primers were designed, which were homologous to the sequences of both erg7 ORF and plasmid pUG66. The cassette was transformed into diploid wild strain INVSc1 by LiAc/SS Carrier DNA/PEG method and then erg7 gene haploid deficient mutant was obtained by homologous recombination. The results of semiquantitative PCR and real-time quantitative PCR revealed that erg7 expression level in erg7 gene haploid deficient mutant is one time lower than that in wild strain. The results of TLC and HPLC showed that the ergosterol content in deficient mutant decreased to 42% of that in wild strain.
Chromatography, High Pressure Liquid ; DNA Primers ; Down-Regulation ; Ergosterol ; metabolism ; Haploidy ; Intramolecular Transferases ; genetics ; Polymerase Chain Reaction ; Saccharomyces cerevisiae ; genetics ; Squalene ; analogs & derivatives ; metabolism

To investigate such physical indexes as hygroscopicity, angle of repose, bulk density, fillibility of compression of mixed powder of directly compressed auxiliary materials and fermented cordyceps powder by using micromeritic study methods. The results showed that spray-dried lactose Flowlac100 and microcrystalline cellulose Avicel PH102 had better effect in liquidity and compressibility on fermented cordyceps powder than pregelatinized starch. The study on the impact of directly compressed auxiliary materials on the powder property of fermented cordyceps powder had guiding significant to the research of fermented cordyceps powder tablets, and could provide basis for the development of fermented cordyceps powder tablets.
Cellulose ; chemistry ; Cordyceps ; chemistry ; Drug Compounding ; methods ; Fermentation ; Lactose ; chemistry ; Powders ; chemistry ; Tablets ; chemistry ; Technology, Pharmaceutical ; methods

Objective To explore the changes of the plasma motilin(MTL)and serum gastrin(GAS)levels and their relationship with course of disease,the severity,the complication of digestive system in neonates with intracranial haemorrhage(ICH).Methods The objects were 26 cases of term newborns with ICH,the healthy control group contained 30 cases of healthy term newborns.Plasma motilin and serum gastrin concentration were measured by radioimmmunoassay in 26 cases of newborns with ICH in the 2 d,3-5 d,7-10 d,and 12-15 d after birth,and compared with the healthy control group.Results Compared with healthy control group,the blood GAS and MTL levels of neonatal ICH group increased dramatically(Pa