Four screening methods, colorimetric assay, blood-plate hemolysis method, surface tension activ- ity and oil spreading technique were introduced to isolate strains producing bio-demulsifiers from 6 different bacteria source samples. The results of various screening methods were evaluated in this paper. Seventeen demulsifying strains were obtained, which are qualified in demulsification test of kerosene model emulsions. Among them, 5 strains showed high demulsifying ability, achieving 70% plus demulsifying ratio within 24 hours. Petroleum-contaminated soil, excess sludge from biological process treating oilfield produced water and sludge from municipal wastewater treatment plant were the best among all tested sources. Due to the determination limit, the colorimetric assay and blood-plate hemolysis method are not competent to screen bio-demulsifiers strains. The measurement of surface tension and oil spreading method were easy but accu- rate methods to isolate bio-demulsifiers strains. Although demulsification test of model emulsion is an effec- tive technique to target strains with the capability of breaking emulsions, it is sophisticated and time-con- suming. Thus it is recommended to use surface tension and oil spreading methods in pre-screening and vali- date the results in demulsification test with kerosene model emulsions.

Aim To observe histopathological changes of hippocampus after acute epilepsy induced by penty-lenetetrazole (PTZ)in rats.Methods Five groups as control group,PTZ-induced 24 hours(h)group,PTZ-induced 72 hours group,PTZ-induced 1 20 hours group and PTZ-induced 1 44 hours group were designed.PTZ (64 mg·kg -1 )was administered with a single intrap-eritoneal injection for generalized tonic-clonic sei-zures in the current experiment.Control and PTZ trea-ted animals were sacrificed after specific time points. Brain was dissected out and then evaluated for neuro-pathological changes using Nissl staining and immuno-histochemical technique.Results In this study PTZ-induced hippocampal neuron status apoptosis occurred at 24 hours and was sustained for 1 44 hours after status epilepticus.Whereas,activated caspase-3 and AIF ap-peared at 24 hours and were sustained for 1 44 hours af-ter status epilepticus.Conclusion The results of this study show that the significant histopathological chan-ges of hippocampus appear in the vicinity of 1 20 hours after intraperitoneal injection of pentylenetetrazole.

Objective To analyze nucleic acid sequence homology of the 6 kb(pYC) plasmid of Yersina. pestis (Y. pestis) isolated from Yurman by searching GenBank. Method The search of sequence similarity was accomplished with BLAST. Results The pYC plasmid sequence had high homology with some genes in nueleotide sequence, such as: 97.1% homology with Shigella sonnei pKYM, 92.1% homology with Haemophilus influenzae(H. influenzae) gene, Salmonella typhi (S. typhi) gene LT2 and plMVSI with 88.2% and 87.2% of homology respectively, Escherichia coli(E, coli) O157:H7 and K-12, ECOR31 with 81.4%, 81.4% and 84.7% of homology respectively. This plasmid ORFs could code for some proteins which were similar with others in GenBank, such as: ORFi and H. paragallinarum replication protein B(47.2%), ORF4 and E. coli hypothetical protein(52.7%), ORF5 and Y. pseudotuberculosis Tile (48.3%), ORF6 and E. coil Pilx5/VirB5-1ike protein (42.3%), Y. enterocolitica TriD protein(38.5%), ORFIO and S. typhimurium LT2, E. coli O157:H7 hypothetical protein(83.1% and 81.9%, respectively), ORF11 and E. coli, damage-inducible protein J(81.4%). Conclusions The pYC plasmid sequence has high homology with a few bacterial genes of Enterobacteriaceac. This plasmid may code for some proteins that are similar with hypothetical protein, damnge-indncible protein, TriD and TilE protein, Pilx5/VirB5-hke protein of Escherichia or Yersinia.

Objective To evaluate the risk of plague occurrence via surveying and analyzing indoor rat density and flea index in natural villages having previous plague experience. Methods During August to September 2007, 30 natural villages experiencing previous plague were selected based on the surveillance data, and then all households were coded with numbers and 20 households in each village were randomly selected via computer. Cages and sticky papers were set in 600 selected households to capture rats and fleas. Rat density, flea prevalence, flea index and median were estimated. Results One hundred thirty-three Rattus flavipectus and 33 Suncus murinus were caught and averaged rat density was 2.8 rats per one hundred cage. nights (166/6000), the median was 5 rats each village. One hundred and one mice infected fleas, flea prevalence on rats was 60.8% (101/166), 296 Xenopsylla cheopis and 48 Leptopsylla segnis were collected. Rat flea index was 2.1 fleas per rat (344/166). A total of 315 dissociated flea was caught, average dissociated flea index was 0.026 fleas per sticky paper (315/11888). The median was 5.5 dissociated fleas per village. Of dissociated fleas, Ctenocephalides felis felis (205) and Xenopsylla cheopis (103) accounted for 97.8% (308/315). The proportion for species of the rat flea and the dissociated flea was different(Fisher test: P < 0.01). The rat flea was significantly associated with the rat density(r = 0.68, P < 0.01), but the dissociated flea was significantly associated with neither the rat density(r = -yield than fried wheat batter(χ2 = 5.59, P < 0.05). Conclusions In these villages having previous plague experience of Lianghe County, Rattusflavipectus was dominant species of indoor rats, Xenopsylla cheopis and Ctenocephalides felis felis were dominant species of rat flea and dissociated flea, respectively. Mengsong, Bangdu, and Tangjiatun village had potential risk of plague emergence.

OBJECTIVETo screen the proteins interacting with inhibitor of differentiation 1(Id1) using yeast two-hybrid analysis in adult human lung cDNA libraries.

METHODSThe coding sequence of Id1 was amplified by PCR and cloned into the bait plasmid. The recombinant bait vector pHybLex/Zeo-Id1 was verified by restriction endonuclease digestion before transformation into the yeast strain EGY48/pSH18-34, which was tested subsequently for reporter genes Leu2 and LacZ activation. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into the yeast strains and screened to obtain Leu2(+) and Leu2(+)LacZ(+) clones, with the false positive clones excluded using positive and negative controls, and the plasmid of the true positive clone was sequenced and blasted for homological analysis.

RESULTSSuccessful construction of pHybLex/Zeo-Id1 was confirmed by enzyme digestion. After transformation of pHybLex/Zeo-Id1 into EGY48/pSH18-34, no specific reporter genes Leu2 and LacZ activation was found. The pHybLex/Zeo-Id1 plasmid and the cDNA library plasmid were sequentially transformed into yeast strain, and 198 Leu(+) clones and 19 Leu(+)LacZ(+) double positive clones were obtained. After elimination of the false positive clones, one true positive clone was obtained, whose plasmid analysis by sequencing and blasting indicated high homology (99.5%, 556/559) to AGGF1 (an angiogenic factor with G-patch and FHA domains 1). AGGF1 expression was confirmed in the true positive yeast cells by Western blotting.

CONCLUSIONAGGF1 is confirmed to interact with Id1 by yeast two-hybrid analysis for screening adult human lung cDNA libraries.

Adult ; Angiogenic Proteins ; genetics ; metabolism ; Cell Proliferation ; Endothelial Cells ; cytology ; metabolism ; Gene Library ; Humans ; Inhibitor of Differentiation Protein 1 ; metabolism ; Lung ; cytology ; Plasmids ; genetics ; Protein Binding ; Two-Hybrid System Techniques

Invigorating blood and dissolving stasis method is a kind of unique therapy of Traditional Chinese Medicine(TCM) treatment, which efficacy has become increasingly prominent in the treatment of ophthalmology.With the further studies of blood stasis and invigorating blood and dissolving stasis therapy, it is widely used in clinical ophthalmology, and get good effects beyond thought, especially when western medicine has no curative effects.It improved the cure rate of fundus oculi disease from the eyelids, conjunctiva, lacrimal sac, vitreous body to the choroid and retina, optic nerve and macula lutea, from surface to fundus, or pathological changes related to inflammation, degeneration, necrosis, atrophy, hyperplasia of fibrous tissue hyperplasia.This paper is aim to explain the definition of invigorating blood and dissolving stasis and make a review of basic research and clinical application about it in several diseases.

AIM To establish an HPLC method for the simultaneous content determination of monocaffeyltartaric acid,forsythiaside A,chicoric acid,phillyrin,arctiin and harpagoside in Waigan Fenghan Granules.METHODS The analysis of 70%methanol extract of this drug was performed on a 30℃thermostatic Agilent Zorbax SB-C18 column(250 mm×4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1%phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 278,330 nm.RESULTS Six constituents showed good linear relationships within their own ranges(r≥0.999 4),whose average recoveries were 96.80%-102.61%with the RSDs of 0.26%-1.93%.CONCLUSION This simple and accurate method can be used for the quality control of Xiao'er Qingyan Granules.

OBJECTIVEPrevious research suggests that dexamethasone (Dex) pretreatment protects neonatal rats against hypoxic-ischemic brain damage (HIBD). Some of the pharmacological effects of baicalin (a traditional Chinese medicine extracted from Scutellaria baicalensis Georgi) are similar to Dex. This study was designed to explore the effect of baicalin on the neuronal apoptosis following HIBD in neonatal rats.

METHODSSix-day-old Sprague-Dawley rats were randomly assigned into Control (without HI), HIBD, Dex-pretreatment and post-treatment, Baicalin-pretreatment and -post-treatment groups. HIBD was induced by ligating the left common carotid artery, followed by exposure to hypoxia. In the pretreatment groups either baicalin (16 mg/kg) or Dex (0.1 mg/kg) was administered to the rats 24 hrs before HIBD; in the post-treatment groups baicalin or Dex was given 30 minutes after HIBD. The rat pups were sacrificed on postnatal day 10, and brain tissues were harvested. Brain water content was determined, morphological changes were observed under a light microscope, and neuronal apoptosis was measured by terminal deoxynucleotidyl transferase mediated dUTP biotin nick end labeling (TUNEL) staining.

RESULTSThe brain water content and the number of apoptotic cells were significantly higher in the HIBD group than those of the Control group (P < 0.05). Both baicalin and Dex pretreatment decreased the brain water content from 88.9 +/- 1.7 % (HIBD group) to 87.4 +/- 0.7% (baicalin) or 87.3 +/- 0.6% (Dex) (P < 0.05) and the number of apoptotic cells were reduced from 251 +/- 28 (HIBD group) to 102 +/- 47 (baicalin) or 75 +/- 26 (Dex) (P < 0.05). Baicalin and Dex post-treatment had no effects on the brain water content and the number of apoptotic cells. Loss and degeneration of neurons could be observed in the HIBD group. Baicalin and Dex pretreatment significantly alleviated neuronal injury, but post-treatment did not.

CONCLUSIONSPretreatment with baicalin, as with Dex, has a protective effect against HIBD in neonatal rats, but baicalin or Dex post-treatment do not reverse the neuronal injuries.

Animals ; Apoptosis ; drug effects ; Body Water ; metabolism ; Brain ; metabolism ; pathology ; Female ; Flavonoids ; therapeutic use ; Hypoxia-Ischemia, Brain ; drug therapy ; metabolism ; pathology ; In Situ Nick-End Labeling ; Male ; Neuroprotective Agents ; therapeutic use ; Rats ; Rats, Sprague-Dawley

AIMTo demonstrate the specific killing of folate receptor (FR)-positive tumor cells can be achieved by folate-targeted penicillin-G amidase (PGA) combined with its prodrug substrate N-(phenylacetyl) doxorubicin (DOXP).

METHODSFolic acid was covalently linked to PGA and folate content value was determined by quantitative UV spectrophotometry. The ability of folate conjugated PGA to hydrolyze DOXP was measured by RP-HPLC. Visual demonstration of uptake by FR (+) HeLa and SKOV3 cells was detected by using FITC labeled folate-PGA and a fluorescence microscopy. The cytotoxicity of DOXP towards the cells in the presence or absence of folate-PGA was assayed by using MTT method.

RESULTSThe folate-PGA has a specific activity of 29. 8 U x mg(-1) (protein). FR selectivity was confirmed by fluorescence microscopy. The combination of DOXP prodrug with folate-PGA generated higher cytotoxicity towards the FR (+) cells than free doxorubicin. The IC50 was 0.72 micromol x L(-1) for HeLa cells and 0.75 micromol x L(-1) for SKOV3 cells, respectively. Further, the enhanced cytotoxicity reduced greatly with the addition of free folic acid.

CONCLUSIONFolate conjugated PGA did not significantly compromise PGA catalytic activity and enabled binding prodrug-activating enzyme PGA to folate receptor expressing cells, and increased the sensitivity of the cells to doxorubicin followed by administration of its prodrug substrate.

Antibiotics, Antineoplastic ; pharmacology ; Carrier Proteins ; metabolism ; Cell Line, Tumor ; Doxorubicin ; analogs & derivatives ; pharmacology ; Drug Delivery Systems ; Female ; Folate Receptors, GPI-Anchored ; Folic Acid ; chemistry ; pharmacology ; HeLa Cells ; Humans ; Inhibitory Concentration 50 ; Ovarian Neoplasms ; pathology ; Penicillin Amidase ; chemistry ; pharmacology ; Prodrugs ; pharmacology ; Receptors, Cell Surface ; metabolism

0.05).Moreover,the growth curves of the two kinds of cells were similar.Con- clusions The cell growth properties of cultured transplanted rabbit SMG are similar to that of normal SMG,the cytobiological charac- teristic of transplanted autologous free rabbit SMG are not changed evidently.