ve To control and minimize the water pollution by nitrogen. Methods The autotrophic amino-oxidizing bacteria in soil were screened by enrichment and incubation. After bacterio-morphological identifica-tion their nitrifying rates were determined. Results All of the 12 screened strains were gram-negative and in the shape of brevi-rod dorminantly. The colonies with a dorminant form of short diameter in round shape, light yellow and smooth formed after 7-day incubation at 28℃ . The nitrifring rates ranged 0.40 mg/ (L?d)~1. 15 mg/ (L?d) . Conclusion The screened 12 strains all presented nitrifying function, some of them could be used to control water environment pollution by nitrogen.

Objective To screen and seperate nitrobacteria that could be used to treat nitrogenous compound pollution.Methods Autotrophic nitrite-oxidizing microorganisms were screened by enrichment culture from soil.After morphological and biological detection,primers were designed to detect the distinctive gene norB of nitrobacteria.The PCR production was se-quenced,and the homology was identified according to the sequence in Genbank.Results One strain of bacterium was screened that could oxidize nitrite.This bacterium was small,light-yellow bacilli.The expected-size DNA fragments could be amplicated by PCR.The sequence of a PCR production was blasted in Genbank and showed99%consistent with norB gene of N hamburgensis.Conclusion It was preliminarily confirmed that the microorganism screened in this study might be nitrobac-terium.

Objective To amplify and sequence the distinctive norB gene of Nitrobacterium. Methods The norB gene was amplified by PCR and was cloned into T-vector and then transformed into E.coli JM109. After white-and-blue selection, positive colonies were sequenced with T7 sequencing primers by Takara Company. Spliced with Vector NTI Suite software, the homologous analysis of sequences were conducted in Genbank through Internet. Results The results showed that all of norB genes obtained from the nitrite-oxidizing bacteria were homologous with the norB gene of Nitrobacterium hamburgensis in Genbank. The homology ranged from 96% to 98%, and there were no frameshift mutations, which guaranteed the correct ORF. Conclusion The obtained genes are norB genes indeed.

Objective To study on glutathione and glutathione-related enzymes of Antarctic ice microalgae.Methods Glutathione(GSH) content,glutathione produce ability(GPA) and glutathione reductase(GR) activity in 24 kinds of Antarctic ice microalgae were assessed using the spectrophotometer methods.Results GSH content of Cyanophyceae B-1 was highest.Total GSH yields of Antarctic ice microalgae Chlamydomonas sp.ICE-L and Berkeleya rutilans GJ01 were in the top two positions.GPA in most Antarctic ice microalgae was higher than that in mesophilic green algae,and only in two ice microalgae was lower than that in Phaeodactylum tricornutum.GR activities of Antarctic ice microalgae Berkeleya rutilans GJ01 and Chlamydomonas sp.ICE-L were greater than that of the control microalgae.Conclusion It is possible that Antarctic ice microalgae will become a new resource of GSH.

Objective To study on quenching free radical substances in antarctic ice alga.Methods Antarctic ice alga Berkeleya rutilans H-15 used as material,complete experiment of preparation free radical scavenging substances was established.All of these include the extraction of active substances by methanol,then pursuing and determination of the separation effect and change of active substances by DPPH and Godin method;the primary separation and purification by silica gel 60 column chromatogram;the farther purification by Sephadex LH column chromatogram and the purity determination of compound by HPLC.The type of compound and functional group of high pure compound were determined by the IR spectrum,HPLC-MS.Results One pure compound with strong activity was obtained from Berkeleya rutilans H-15,and it was confirmed to belong to phenol derives by analysis of IR spectrum and HPLC-MS.Conclusion It was completely feasible to find and acquire quenching free radical substances in antarctic ice alga,which initiated a new way to obtain natural antioxidant active substances from antarctic microorganism.

Objective To study on algicidal bioactive substances from mangrove bacteria and its algicidal effect on red tide algae Alexandriam tamarense.Methods A red algicidal bioactive substance in Guangxi mangrove bacteria was isolated and purified by Silica-gel Column Chromatography,and its structure was determined on the basis of UV,IR,1H NMR,13C NMR and HREIMS spectroscopic analyses and comparison with the literatures.Its algicidal affect was primarily studied in this paper.Results and Conelusion The results showed that the red algicidal bioactive substance was prodigiosin,which had an inhibitory effect on the cells growth of Alexandrium tamarense,and the inhibitory effect increased with the increase of prodigiosin concentration.The prodigiosin stimulated the production of MDA and reduced the content of sulfhydryl group.In addition,plasma membrane permeability of leaked cell solution was increased.These indicated the extinguishing mechanism: the cell membrane was damaged followed by the inhibition of the cell growth.

A yeast strain Rhodotorula sp.NJ-298,which was isolated from antarctic sea-ice,could produce carotenoids pigment,and the total production was 1986?g?g~(-1).UV-visible spectrum and LC/MS analysis indicated astaxanthin was the major component of the carotenoids,and HPLC analysis showed its content accounted for 81.9 % of total carotenoids productoin.Maximum astaxanthin production 6506 ?g?L~(-1) was obtained when the yeast was grown at steady-state phase.Optimum temperature for the strain grow was at 8℃,and sodium acetate and peptone were the best carbon and nitrogen sources for the astaxanthin production.Moreover,the pigment had strong activity of scavenging free radicals,and the IC_(50) was 0.40 ?g?mL~(-1) by DPPH assay.

OBJECTIVETo assess the value of ultrasonic radio frequency analysis technique for noninvasively evaluation of carotid artery intima-media thickness (IMT) and elasticity in patients with dyslipidemia.

METHODSRadio frequency quality intima-media thickness (RFQIMT) technique and radio frequency quality arterial stiffness ((RF)QAS) technique were used for measurement of IMT and pulse wave velocity (PWV) of the left carotid artery in 86 patients with dyslipidemia and 60 healthy volunteers.

RESULTSApart from the patients' age, body mass index, systolic blood pressure, diastolic blood pressure, pulse pressure, total cholesterol, triglyceride, low density lipoprotein, high density lipoprotein, fasting blood glucose, IMT and PWV differed significantly between the control and case groups. IMT was positively correlated with PWV (r=0.521, P<0.05). In patients with IMT<1.0 mm (n=81), PWV differed significantly between patients with different number of other atherogenic risk factors (P=0.004), but IMT showed no such variation (P=0.079). In patients with more than 3 and those with 2 other risk factors, PWV increased significantly as compared with those with one risk factor (P=0.002). Multiple linear regression analysis showed that systolic blood pressure, age and fasting blood glucose were independently associated with PWV.

CONCLUSIONUltrasonic radio frequency analysis technique allows accurate measurement of IMT and arterial elasticity, and may thus become a valuable method for evaluating early structural and arterial functional change of the carotid artery.

Adolescent ; Adult ; Aged ; Carotid Arteries ; diagnostic imaging ; physiopathology ; Carotid Intima-Media Thickness ; Case-Control Studies ; Dyslipidemias ; diagnostic imaging ; physiopathology ; Elasticity ; Female ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo establish a high performance liquid chromatography method for the determination of tetrahydropulmatine content in treatment of winter's diseases in summer path.

METHODThe analysis was performed on the Hypersil ODS2 column (4.6 mm x 250 mm, 5 microm) with methanol and 0.1% phosphoric acid solution (adjusted pH to 6.28 with triethylamine) as mobile phase at 63:37. The flow rate was 1.0 mL x min(-1) and the UV detection wavelength was 280 nm.

RESULTThe linear range of tetrahydropulmatine was obtained from the range of 2.08 - 49.92 mg x L(-1) (r = 0.999 8). The average recovery was 97.85% with RSD 2.49%.

CONCLUSIONThis method is simple, reliable and can be used for the determination of tetrahydropulmatine in this preparation.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Reproducibility of Results

The aim of this study is to quantify the 146S antigen in foot-and-mouth disease virus (FMDV) inactivated vaccine by size-exclusion chromatography (SEC). The analysis was performed on a TSKgel G4000SWXL column (7.8 mm×30 cm), with a pH 7.2 buffer salt system as the mobile phase. The flow rate was 0.6 mL/min, the injection volume was 100 μL and the detection wavelength was 259 nm. The calibration curve was established by using purified inactivated FMDV (serotype O) 146S antigen; 3 batches of vaccine formulated by inactivated antigen solution were tested to verify the accuracy, reproducibility, specificity and tolerability of the method. At last 16 batches of vaccine were determined by the SEC method. Results showed a good linearity between peak area and concentration of 146S antigen in the range between 0.56 and 67.42 μg/mL (R2=0.996, n=10), and the average recovery rate of 146S antigen in the 3 batches of vaccine formulated in lab were 93.6% (RSD=2.7%, n=3), 102.3% (RSD=2.6%, n=3), and 95.5% (RSD=5.1%, n=3). The method was proved accurate and reliable with good reproducibility (RSD=0.5%, n=6), and applied to determine 16 batches of the commercial FMDV vaccine. According to the above results, the SEC method is high effective for 146S antigen quantify in the inactivated FMDV vaccine and would provide strong support for the vaccine quality control.