Using of clearing zones on pachyman agar medium, there are 217 plant endophytic actinomycetes, producing ?-1,3-glucanase were screened. 45.6% of the strains produced ?-1,3-glucanase, in which the strains from cucumber are up to 38. The percentages of endophytic actinomycetes from different hosts produceing ?-1,3-glucanases were different. The percentage of the strains in Rhizoma Polygonatum produced ?-1,3-glucanases is the highest, up to 88.9%. The Inhibited effects of plant endophytic actinomycetes which produced extracellular ?-1,3-glucanases on mycelium growth of Sclerotinia sclerotiorum were detected in vitro. Cucumber endophytic actinomycete gCLA4 strain was screened out from 99 isolates, which can strongly inhibit the growth of S. sclerotiorum. The optimal ?-1,3-glucanases fermentation conditions of strain gCLA4 were investigated, they were pachyman 0.2%, peptone as nitrogen, pH 7～8 for 5 days. The ?-1,3-glucanases of strain gCLA4 had some inhibiting efficiency on 13 plant pathogens, in which inhibiting efficiency to Botryosphaeria dothidea was the strongest.

Background The current epidemiology study had shown the prevalence of age and sex adjusted dry eye was higher in patients with diabetes than population without diabetes.Further researches demonstrated that the tear film disturbance is common after the phacoemulsification or photocoagulation in the eyes of diabetic patients.Objective The aim of this study was to characterize the clinical features of tear film instability in diabetes patients.Methods One hundred and sixty-two patients with tear-film abnormality referred to Tongren Eye Center from January 1,2010 to September 1,2010 underwent questionnaire about diabetes and other diseases,BUT,Schirmer test.Tear film instability was diagnosed as abnormality of either Schirmer test or BUT and showed as M ( Q25,Q75 ).The right eyes of 162 dry eye patients meeting with the including criteria were enrolled.The patients were assigned to two groups according to with ( 80 patients) or without ( 82 patients) diabetes mellitum.DEQ questionnaire were scored.The percentage of cases with meibomain gland abnormal score ＞ 1 was calculated.Mann-Whitney U analysis and Chisquare analysis were used to compare the difference between the two groups.Results The Schirmer test in diabetic group was 8 ( qualities:5,9 )mm and was longer than 6 ( qualities:5,7 ) mm in non-diabetic patients ( U =2452,P =0.00).The result of BUT test was 3 ( qualities:2,4 ) seconds in diabetic patients and was shorter than 4 (qualities:3,5) seconds in non-diabetic patients( U=2104,P＜0.01 ).The DEQ score of diabetic patients was 15 ( qualities:1 0,19,which was less than21 ( qualities:19,23.25 ) in non-diabetic patients.51.2 ％ ( 41/80 ) diabetic participants and 32.9％ (27/82) nondiabetic participants appeared meibography ( grade larger than 1 ) (x2 =16.07,P=0.00).The percentages of dry eyes were 51.2％ (41/80) and 93.9％ (77/82) respectively in diabetes and nondiabetes groups(x2 =37.24,P＜0.01 ).No significant correlation was found between the diabetes course and DEQ score or meibography( r =0.16,P =0.16 ; r =0.10,P =0.36 ).Conclusions Diabetes patients with tear film instability have longer Schirmer test results,shorter BUT,more severe meibomain glands damage and lower DEQ scores.The dry eye symptom is lack in the diabetic patients though appearing the tear film and meibomain glands damage.Therefore,more attention should be given to ocular surface health in diabetes patients.

AIMThe liposome/water partition coefficients of salmeterol and budesonide between aqueous phase and liposomes were determined and the factors that influence their partition coefficients were studied, the mechanism of interaction between the two drugs and phospholipid bilayer was elucidated.

METHODSThe liposome/water partition coefficients of the two drugs were determined by equilibrium dialysis technique. The change of the partition coefficients of the two drugs along with liposome composition and medium was also studied.

RESULTSThe partition coefficients of the two drugs decreased with the increase of cholesterol content and saturation of phospholipid used. The liposome/water partition coefficient of salmeterol increased with the increase of liposome surface negative charge, medium pH and ionic strength, while the liposome surface charge, medium pH and ionic strength had no distinct effect on the liposome/water partition coefficient of budesonide.

CONCLUSIONThe liposome/water partition coefficient of drug was affected by the type, saturation of phospholipid used in liposome preparation, the cholesterol content and surface charge of liposome, as well as the pH and ionic strength of medium also have effect on the liposome/water partition coefficient of drug. Accordingly, in order to reflect the actual partition of drug in biological membrane, the determination condition including liposome composition and medium should be similar to the biological membrane.

Albuterol ; analogs & derivatives ; chemistry ; Budesonide ; chemistry ; Cholesterol ; chemistry ; Drug Carriers ; Hydrogen-Ion Concentration ; Ions ; Liposomes ; Membranes, Artificial ; Phospholipids ; classification ; Salmeterol Xinafoate ; Water ; analysis

To explore the mechanism of the absorption enhancement of borneol, the effect of borneol on the intestinal absorption and the pharmacokinetics of tetramethylpyrazine phosphate after oral administration were investigated. In situ intestinal recirculation was performed to study the effect of various concentrations of borneol on the absorption of tetramethylpyrazine phosphate at duodenum, jejunum, ileum and colon. After oral administration of tetramethylpyrazine phosphate, the mixture of tetramethylpyrazine phosphate and borneol and the mixture of tetramethylpyrazine phosphate and verapamil in rats, the concentrations of tetramethylpyrazine phosphate in plasma were determined by RP-HPLC at predesigned time. The pharmacokinetic parameters were compared based on the results of the three animal experiments, and analyzed with software program 3p97. The result showed that tetramethylpyrazine phosphate could be absorbed at all of the four intestinal segments with increasing absorption amount per unit as follows: colon > duodenum > jejunum > ileum, but without saturation, which demonstrated that tetramethylpyrazine phosphate was absorbed via simple diffusion. Borneol could enhance the intestinal absorption of tetramethylpyrazine phosphate, however, not in proportion. There was no obvious difference between the test group and the control group when 10 microg x mL(-1) borneol was added (P > 0.05), while when the concentration comes to 25 microg x mL(-1) and 50 microg x mL(-1), significant differences were observed (P < 0.05). Borneol and verapamil did enhance the bioavailability of tetramethylpyrazine phosphate after oral administration in rats. The enhancing effect of borneol showed only when the concentration came to a certain level but with no specific sites existed in the intestine. One of the mechanisms of borneol on the enhancing effect on absorption of tetramethylpyrazine phosphate might be the inhibition of the metabolism of CYP 3A and exocytosis of P-gp.
Animals ; Biological Availability ; Bornanes ; pharmacokinetics ; Herb-Drug Interactions ; Intestinal Absorption ; drug effects ; Male ; Pyrazines ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

To optimize indices of molecular identification for authentication of Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, four indices, including sequence similarity, specific positions, genetic distance and phylogenetic tree, were compared based on trnL-trnF sequences. Total DNA was extracted from Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix, and trL-trnF sequences were amplified and sequenced. Sequence similarity was calculated by BLAST analysis. Specific positions were compared by DNAman software. Genetic distance and phylogenetic tree were analyzed by Mega software. The results showed that the inter-specific and intra-specific similarity of P. ginseng and P. quinquefolius respectively was 100% and 99. 6%. There were four specific positions at G153A, T463A, C732G and T818C. The inter-specific genetic distance (0) of trL-trnF sequences was lower than intra-specific genetic distance (0. 004). P. ginseng can be distinguished from P. quinquefolius based on the phylogenetic tree. It is concluded that Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix can be authenticated by identification indices of sequence similarity, specific positions, genetic distance and phylogenetic tree. Index of specific positions based on trnL-trnF sequences is the most efficient index to authenticate Ginseng Radix et Rhizoma and Panacis Quinquefolii Radix.
Chloroplasts ; genetics ; DNA Barcoding, Taxonomic ; methods ; Panax ; classification ; genetics ; Phylogeny ; Plant Proteins ; genetics ; Rhizome ; classification ; genetics

Glucose ; Humans ; Hypothalamo-Hypophyseal System ; Lycium ; Pituitary-Adrenal System ; Resistance Training

The fruit of Lycium ruthenicum is a common folk medicine in China. Now it is popular for its antioxidative effect and other medical functions. The adulterants of the herb confuse consumers. In order to identify a new adulterant of L. ruthenicum, a research was performed based on NCBI Nucleotide Database ITS Sequence, combined analysis of the origin and morphology of the adulterant to traceable varieties. Total genomic DNA was isolated from the materials, and nuclear DNA ITS sequences were amplified and sequenced; DNA fragments were collated and matched by using ContingExpress. Similarity identification of BLAST analysis was performed. Besides, the distribution of plant origin and morphology were considered to further identification and verification. Families and genera were identified by molecular identification method. The adulterant was identified as plant belonging to Berberis. Origin analysis narrowed the range of sample identification. Seven different kinds of plants in Berberis were potential sources of the sample. Adulterants variety was traced by morphological analysis. The united molecular identification-origin-morphology research proves to be a preceding way to medical herbs traceability with time-saving and economic advantages and the results showed the new adulterant of L. ruthenicum was B. kaschgarica. The main differences between B. kaschgarica and L. ruthenicum are as follows: in terms of the traits, the surface of B. kaschgarica is smooth and crispy, and that of L. ruthenicum is shrinkage, solid and hard. In microscopic characteristics, epicarp cells of B. aschgarica thickening like a string of beads, stone cells as the rectangle, and the stone cell walls of L. ruthenicum is wavy, obvious grain layer. In molecular sequences, the length of ITS sequence of B. kaschgarica is 606 bp, L. ruthenicum is 654 bp, the similarity of the two sequences is 53.32%.
Berberis ; classification ; cytology ; genetics ; China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; chemistry ; genetics ; DNA, Ribosomal Spacer ; chemistry ; genetics ; Drug Contamination ; Drugs, Chinese Herbal ; isolation & purification ; standards ; Lycium ; classification ; cytology ; genetics ; Medicine, Chinese Traditional ; Phylogeny ; Sequence Analysis, DNA ; Species Specificity

The tandem mass spectrum of apigenin-6,8-C-di-glucoside( 1) and apigenin-6-C-glucose-8-C-rhamnoside( 2) were obtained by high resolution electrospray ionization mass spectrometry( HR-ESI-MS/MS) in both positive and negative ion modes. The elemental composition of each ion was determined according to its accurate mass-to-charge,hence,the fragmentation pathways of each compound were proposed in both negative and positive ion modes. Comprehensive analysis of each ion and its proposed fragmentation pathways of the two compounds was initially conducted in both negative and positive ion mode HR-ESI-MS/MS to explore the diagnostic ions for flavone-6,8-C-di-glycosides and the characteristic ions for each compound and their cleavage rules. The results showed that a family of fragmentation ions with m/z 353,325,311,297 in ESI(-)-MS and m/z 355,325,307,295 in ESI( +)-MS could be the diagnostic ions of flavone-6,8-C-di-glycoside,and characteristic neutral loss could be assigned to glycosyl substitution,for example,neutral losses of C_4H_8O_4( 120),C_3H_6O_3( 90),C_2H_4O_2( 60) for glucoside substitution while neutral losses of C_4H_8O_3(104),C_3H_6O_2( 74),C_2H_4O( 44) for rhamnoside substitution. Furthermore,only one H_2O loss from mother ion( [M-H]-) was observed for 1 & 2 in ESI(-)-MS while five to six H2 O loss from mother ion( [M+H]+) was observed for 1 & 2 in ESI( +)-MS to produce a family of ions by subsequent loss of H_2O,which could be applied for glucosyl difference. The flavone-6,8-C-di-glycosides in both ESI( +)-MS and ESI(-)-MS showed the cleavage similarity at sugar substitutions. However,there were much more differences by the fragmentation pathways and neutral losses between ESI( +)-MS and ESI(-)-MS as following,hyperconjugation ions by subsequent loss of H_2O from precursor ions of flavone-6,8-C-di-glycosides in ESI( +)-MS were not observed in ESI(-)-MS; the subsequent neutral loss of CH_2O in ESI( +)-MS were rarely observed in ESI(-)-MS; the loss of CO only happen at C-ring of flavone ESI( +)-MS other than glycosyl position in ESI(-)-MS; the C4-chain neutral loss of flavone-6,8-C-di-glycosides happened at 8-C-glycosyl position other than at 6-C-glycosyl position. The above cleavage rules and diagnostic ions of ESI( +)-MS were successfully applied for the structure identification of 4 flavone-6 C,8 C-diglycosides from the stem extract of Dendrobium officinale as vicenin Ⅱ,vicenin Ⅰ,isoschaftoside,schaftoside as well as one flavone-O-glysoside named rutin,which were supported by ESI(-)-MS data as well.
Flavones/chemistry* ; Glycosides/chemistry* ; Ions ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry

The purpose of this study was to establish real-time based methods for detection of NPM1 gene mutation in acute myeloid leukemia (AML). Primers/probes were designed according to the clustered region of NPM1 mutations on exon 12. Two real-time PCR assays, including high resolution melting curve (HRM) and allele-specific PCR (AS-PCR), were developed and clinically evaluated with 89 AML samples, which were parallelly detected by capillary electrophoresis (CE) and sequencing. The results showed that a total of 17 mutation-positive samples were detected, including type A (15 cases), type B (1 case) and type Nm (1 case). HRM assay could detect all mutant types, and the analytical sensitivity was around 5%. In contrast, AS-PCR assay detected only 95% mutant types, but its sensitivity was as high as 0.01%. It is concluded that considering the characteristics of each method as well as the clinical evaluation results, HRM may be used for screening of NPM1 mutations at diagnosis, while the AS-PCR can be used for the MRD quantification during follow-up.
Alleles ; DNA Mutational Analysis ; Electrophoresis, Capillary ; methods ; Genome ; Humans ; Leukemia, Myeloid, Acute ; diagnosis ; genetics ; Mutation ; Neoplasm, Residual ; diagnosis ; Nuclear Proteins ; genetics ; Plasmids ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

In order to minimize the residual tert-butyl alcohol (TBA) level in cyclodextrin complex prepared by freeze drying TBA/water cosolvent system, the formulation and lyophilization procedure that may influence the residual TBA was studied. Residual TBA in freeze dried cyclodextrin complex was determined by gas chromatography. The significant formulation and processing factors that influence residual TBA were identified by adjusting the initial TBA concentration in cosolvent, selecting cyclodextrin type (beta-cyclodextrin or hydroxypropyl beta-cyclodextrin), changing sample volume in flasket, altering freezing mode (fast freezing or slow freezing) and modifying the duration of secondary drying. The results show that the amorphous cyclodextrin material (hydroxypropyl beta-cyclodextrin), initial low TBA concentration in cosolvent and fast freezing would lead to high TBA residue in cyclodextrin complex, annealing was effective in reducing the residual TBA. The duration of secondary drying had no distinct effect on residual TBA. It is concluded that in order to reduce residual TBA in cyclodextrin complex prepared by lyophilization monophase solution, the initial TBA concentration in cosolvent should be higher than the crystal formation concentration, the appropriate cyclodextrin type and freeze drying processing should be choosen.
2-Hydroxypropyl-beta-cyclodextrin ; Anti-Inflammatory Agents ; chemistry ; Budesonide ; chemistry ; Chromatography, Gas ; Drug Residues ; analysis ; chemistry ; Freeze Drying ; methods ; Solvents ; chemistry ; Water ; chemistry ; beta-Cyclodextrins ; chemistry ; tert-Butyl Alcohol ; analysis ; chemistry