Methods of ELISA, nonradioactive molecular hybridiz ation and RT-PCR were applied in the detection of rice grassy stunt virus (RGSV ). The detection sensitivity of indirect ELISA using antiserum against fusion p rotein GST-NC was 1 mg of infected leaves or 84 ng of purified virus. The metho d of dot hybridization using NC, a DIG-labelled DNA probe was 50 μg diseased l e aves, or 6 ng purified preparations. The detection endpoint of RT-PCR was 10 μg diseased leaves, or 2 ng purified virus preparation. Comparisons of sensitivit y and maneuverability were made among these methods.

Benzene ; analysis ; Chemical Industry ; Chromatography, Gas ; methods ; Humans ; Occupational Exposure ; analysis ; Toluene ; analysis ; Trichloroethylene ; analysis ; Xylenes ; analysis

SMART-RACE was performed after isolating the total RNA of Armillariella tabescens to amplify the full-length cDNA of arabinosidase (GenBank Accession No. AJ620046). Bioinformatics analysis was used to analyze the code frame of arabinosidase, to predict its structure and function. Recombinant plasmid pPIC9-AF was constructed and then electroporated into methylotrophic yeast Pichia pastoris GS115. The secreted 6 ? His fusion protein was purified to analyze its enzymology property. This arabinosidase had high activity at 30-35℃ under acid condition, and was stable within wide range of pH and temperature. It maintained about 80% activity at the range of pH4. 0-8.0 and 20-40℃,wider than many other cloned arabinosidase. So it was worthy to go step further to study this enzyme, and recombinant expression provided a chance of highly expressing arabinosidase.

Objective To compare BACTEC Plus aerobic and anaerobic bottles with BacT/ALERT FA aerobic bottles and FN anaerobic bottles in the ability of detecting simulated bacteremia specimens.Methods The 202 pairs of specimens were composed of 5ml sterile blood and defined loads of microorganisms.112 pairs of specimens in them also contained defined doses of antibiotics to simulate the patients undertaking antibiotic therapy.Time-to-detect(TTD)and positive percentages were evaluated in four groups,including aerobic bottles detecting aerobic bacteria,anaerobic bottles detecting anaerobic bacteria and facultative anaerobic bacteria,and aerobic bottles contained antibiotics detecting aerobic bacteria.Results The positive percentages of two kinds of aerobic bottles were both 100%.For the specimens with bacterial concentration of 10~2 and 10~3 CFU/ml,average TTDs of BACTEC Plus aerobic bottles[(13.69?3.74)h,(11.54?2.87)h]were faster than those of BacT/ALERT FA bottles [(16.76?5.62)h,(14.47?4.30)h;t=-5.674,-7.294,P

In order to construct an integrated DNA barcoding database for identifying Chinese animal medicine, the authors and their cooperators have completed a lot of researches for identifying Chinese animal medicines using DNA barcoding technology. Sequences from GenBank have been analyzed simultaneously. Three different methods, BLAST, barcoding gap and Tree building, have been used to confirm the reliabilities of barcode records in the database. The integrated DNA barcoding database for identifying Chinese animal medicine has been constructed using three different parts: specimen, sequence and literature information. This database contained about 800 animal medicines and the adulterants and closely related species. Unknown specimens can be identified by pasting their sequence record into the window on the ID page of species identification system for traditional Chinese medicine (www. tcmbarcode. cn). The integrated DNA barcoding database for identifying Chinese animal medicine is significantly important for animal species identification, rare and endangered species conservation and sustainable utilization of animal resources.
Animals ; DNA Barcoding, Taxonomic ; methods ; Databases, Nucleic Acid ; Eukaryota ; classification ; genetics ; Medicine, Chinese Traditional

Though all the marketed drugs of dipeptidyl peptidase IV inhibitors are structurally different, their inherent correlation is worthy of further investigation. Herein we rapidly discovered a novel DPP-IV inhibitor 8g (IC50 = 4.9 nmol.L-1) which exhibits as good activity and selectivity as the market drugs through scaffold hopping and drug splicing strategies based on alogliptin and linagliptin. This study demonstrated that the employment of classic medicinal chemistry strategy to the marketed drugs with specific target is an efficient approach to discover novel bioactive molecules.
Dipeptidyl-Peptidase IV Inhibitors ; chemical synthesis ; chemistry ; Drug Design ; Drug Discovery ; methods ; Humans ; Hypoglycemic Agents ; chemical synthesis ; chemistry ; Linagliptin ; chemical synthesis ; chemistry ; Molecular Structure ; Piperidines ; chemical synthesis ; chemistry ; Structure-Activity Relationship ; Uracil ; analogs & derivatives ; chemical synthesis ; chemistry

Objective To investigate the in vitro antimicrobial activity of ampicillin-sulbactam,clindamycin and cefoperazone a- gainst common clinical isolates.Methods MICs of these three antibiotics were determined by E test.Results Ampicillin-sulbae- tam showed inhibition rate of 100% for methicillin susceptible S.aureus (MSSA),E.faecalis,Group A Streptococcus,H. influenzae,penicillin-susceptible S.pneumoniae (PSSP),M.catarrhalis and anaerobes (including 10 strains of Bacteroid spp.,2 strains of P.aches,1 strain of E.lentum,1 strain of Fusobacterium innocuum,and 2 strains of Peptostreptococcus spp.).Ampicillin-sulbactam was active against 86.7% of extended-spectrum?-lactamase non-producing K.pneumoniae (NES- BL-KPN) and 53.3% of non ESBL-producing E.coli (NESBL-ECO).Ampicillin-sulbactam only inhibited 23.3% of A.bau- mannii isolates and 25.0% of E.faecium isolates.For MSSA,anaerobes,PSSP and group A Streptococcus,about 60.0%, 31.2%,30.0% and 10.0% of the isolates were susceptible to clindamycin,respectively.For MSSA,NESBL-ECO and NES- BL-KPN,96.7% to 100% of the isolates were susceptible to cefoperazone.About 40.0% of P.aeruginosa isolates were sus- ceptible to cefoperazone.No A.baumannii isolate was susceptible to cefoperazone.Conclusions The ampicillin-sulbactam has good antimicrobial activity against MSSA,E.faecalis,Group A Streptococcus,NESBL-KPN,H.influenzae,PSSP,M.ca- tarrhalis and anaerobes.In this study,clindamycin is active against 60% of MSSA isolates.Most of other species are relatively resistant to clindamycin.Cefoperazone shows good activity against MSSA,NESBL-ECO,and NESBL-KPN.These three anti- microbial agents can be used as empirical treatment for community-acquired infections.

Animals ; Chromatography, Thin Layer ; Dichlorvos ; blood ; Dimethoate ; blood ; Insecticides ; blood ; Methyl Parathion ; blood ; Mice ; Organothiophosphorus Compounds ; blood

OBJECTIVETo investigate the effects of SOM230, a new somatostatin analogue, on the proliferation of hepatocellular carcinoma (HCC) cell line HepG2 in vitro and in vivo, and explore the mechanism underline the necrosis of tumors.

METHODSMTT, TdT-mediated dUTP nick end labeling assay (TUNEL) and flow cytometric assay were used to measure the effects of SOM230 on the proliferation and apoptosis of HCC HepG2 cells. Nude mice bearing HCC xenografts of the HepG2 cell line were treated with SOM230 (100 microg/kg/d subcutaneously injection) and saline as a control for eight weeks. The mass and percentage of necrotic volume of the HCC xenografts in nude mice were determined. Western blot was used to detect SSTR2 in HCC xenografts. Immunohistochemical method was used to detect the expression sites of SSTR2 and VEGF in HCC xenografts. ELISA was used to detect the levels of TNFalpha.

RESULTSNo proliferation and apoptosis of HepG2 cells were induced by SOM230 in vitro (F = 0.16, P more than 0.05). The percentage of necrotic volume in SOM230 were significantly higher than that of control group (73.4%+/-7.0% vs 30.2%+/-14.0%, t = -8.02, P more than 0.01). SSTR2 was expressed in blood sinus of HCC xenografts in nude mice. There was no significance difference in the level of SSTR2 expression between SOM230 group and saline treated group. VEGF expression in xenografts was down-regulated by SOM230 treatment. SOM230 treatment did not affect the level of TNFalpha in HCC xenografts (t = -0.24, P more than 0.05).

CONCLUSIONSSOM230 can induce massive necrosis of HCC xenografts only after the blockage of blood flow through down-regulation of VEGF mediated by SSTR2.

Animals ; Antineoplastic Agents ; administration & dosage ; pharmacology ; Carcinoma, Hepatocellular ; blood supply ; metabolism ; pathology ; Cell Proliferation ; drug effects ; Disease Models, Animal ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Hep G2 Cells ; Humans ; Immunohistochemistry ; Injections, Subcutaneous ; Liver Neoplasms ; blood supply ; metabolism ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Random Allocation ; Receptors, Somatostatin ; metabolism ; Somatostatin ; administration & dosage ; analogs & derivatives ; pharmacology ; Vascular Endothelial Growth Factor A ; metabolism ; Xenograft Model Antitumor Assays

AIM: To screen TIGR/myocilin gene (MYOC) mutation in high myopic Chinese children with family history.METHODS: Gene sequencing was performed in exon 3 of the TIGR gene in high myopic Chinese Children. The coding sequence of TIGR exon 3 was screened by capillary electrophoresis sequencing. The sequence alterations were analyzed by bioinformatics.RESULTS: TIGR gene mutation was not found in high myopic patients and normal controls group.CONCLUSION: No identified gene mutation is found in TIGR gene in high myopic Chinese children.