ObjectiveTo express and purify the P2 domain of 5 genotypes(GⅠ.1,GⅡ.2,GⅡ.3,GⅡ.4,and GⅡ.17)of norovirus(NV)in prokaryotic expression system and prepare their antisera.MethodsThe bioinformatics method was used to analyze the sequence conservation of P2 domains of different genotypes of NV. Then the P2 DNA sequence was synthesized and subcloned into pET24a vector to construct the recombinant prokaryotic expression plasmid,which was transformed into E.coli BL21(DE3)competent cells and induced by IPTG. 30 female BALB/c mice were immunized with the purified recombinant P2 protein to prepare antiserum. The titer and cross reaction of antiserum were detected by ELISA,the specificity was detected by Western blot,and the recognition and binding ability to wild-type NV were analyzed by dotblot.ResultsThe expressed recombinant P2 proteins showed purity of over 90% with the relative molecular mass of about15 000. The expression levels of P2 protein of GⅠ.1,GⅡ.2,GⅡ.3,GⅡ.4,and GⅡ.17 were 60,26. 4,19. 8,16. 3 and33. 3 mg/L,respectively. The antiserum titers against five genotypes of NV P2 antigens were all over 1∶128 000,which well recognized their corresponding P2 antigens,while showed no interaction with unrelated P2 antigens and no crossreaction with recombinant human group A rotavirus VP7 antigen and recombinant Helicobacter pylori uroenzyme antigen,and had good recognition and binding ability to wild-type NV.ConclusionFive genotypes of NV P2 antigens epidemic in China were successfully expressed and purified by prokaryotic expression system. The prepared antisera had high titer and good specificity,which laid a foundation of the establishment of rapid and on-site diagnosis technology of NV.

OBJECTIVE: To test the practicability of the solid-state fermentation for medicinal fungi by fermenting Ganoderma lucidum with Radix Astragali containing medium. METHODS: Ganoderma lucidum was fermented in ordinary medium, drug-containing medium (containing Radix Astragali) and selenium-rich drug-containing medium respectively. The polysaccharide contents of fermentation products from the three kinds of culture media were tested at different time, and the changes were compared. RESULTS: The polysaccharide contents of fermentation products from the three kinds of culture media were 4.65%, 3.76% and 4.50% respectively and their relative standard deviation were 1.61%, 1.99% and 1.86% respectively. By observing the changes of the contents of polysaccharide, protein and total saponin in fermentation products from the drug-containing medium at different time, it was found that the 28th fermentation day was the time when secondary metabolism was most active, and it should be the fermented terminal point. CONCLUSION: The fermentative combination of Ganoderma lucidum and Radix Astragali is practicable.

Objective To prepare and purify the polysaccharides in Shuanqian fungal substance (FS);To study its monosaccharide composition and property. Methods Strychnos nux-vomica was under a thirty-day solid fermentation through Trametes cinnabarina. Then crude polysaccharide was got after the process of drying, smashing, water boiling, condensing, decoloring, removing protein and ethanol precipitation. The content of polysaccharides in Shuanqian FS was determined by phenol-sulfuric acid method. Dialysis and gel chromatography method was used to purify the crude polysaccharide. The purified composition was analyzed by acid hydrolysis and thin layer chromatography. Results The content of crude polysaccharide was 47.68%and yield was 10.52%. It showed that polysaccharide in Shuanqian FS contained glucose and galactose, but no nucleic acid and protein. Conclusion Polysaccharide in Shuanqian FS was an offwhite powdery heteropolysaccharide, which contained glucose and galactose.

Medical staff must pay attention to the following ethical questions during the salvage of SARS patients: (1)respecting SARS patients' dignity and personality; (2)respecting SARS patients' right of hospitalizing equally ; (3)adjusting SARS districts' ethical relationship between patients and doctors; (4)respecting SARS patients' informed consent right; (5)not ignoring the caring of SARS patients until the last;etc.

AIM: To analyze the total chrysophanol contents in the processed seeds of Cassia Tora and the contents of chrysophanol in the crude and processed seeds from different districts are compared. METHODS: HPLC with a Kromasil C 18 column was used and MeOH 0.1%H 3PO 4(85∶15) used as the mobile phase. The flow rate was 1.0 mL?min -1 and detection wavelength was 254 nm. RESULTS: The contents of total chrysophanol was 0.196 4% and the average recovery was 96.95%. The content of free chrysophanol was increased after processing, total and combined chrysophanol were decreased. CONCLUSION: The method is reliable of controlling the quality of Cassia obtusifolia and can be used for further studies in the processing of Cassia obtusifolia.

The viscosity of mucopolysaccharides in Holothuria scabra Jaeger was related to temperature, time, concentration of mucopolysaccharides and H 2O 2 during the decoloration, which was found through the experiment, in that the rise of temperature, elongation of time, decrease of concentration of Mucopolysaccharides and increase of that of H 2O 2 all lead to the decrease of the viscosity. Within a certain limit, the activity of mucopolysaccharides rises with the viscosity decrease in the experiment of acute cerebral ischemia in mice. So that it is important controlling the craft in the mucopolysaccharides production.

OBJECTIVE:To establish a method for simultaneous determination of puerarin,berberine hydrochloride and ba-icalin in Erxieting syrup. METHODS:HPLC was performed on the column of Kromasil-C18 with mobile phase of methanol-0.2%Phosphoric acid solution (gradient elution) at flow rate of 1.0 ml/min,column temperature was 30 ℃,detection wavelength was 275 nm,and the volume injection was 10 μl. RESULTS:The linear was 10.43-83.44 μg/ml for puerarin,10.71-85.68 μg/ml for ber-berine hydrochloride and 12.58-100.64 μg/ml for baicalin(r≥0.999 3);RSDs of precision,stability and reproduciblity tests were lower than 2%;recoveries were 98.25%-100.87%(RSD=0.92%,n=6),99.41%-101.40%(RSD=0.68%,n=6)and 98.13%-99.76%(RSD=0.57%,n=6),respectively. CONCLUSIONS:The method is simple,reliable,can be used for the quality control of Erxiet-ing syrup.

Objective:To optimize the extraction procedure of total alkaloid in Radix Aconiti.Methods:The orthogonal design(L9(34) ) was carried out for the optimum extraction conditions guided by yield of the extract and the content of aconitine,hypaconitine and mesaconitine determined with RP-HPLC method,the mobile phase was MeOH-H2O2-CHCl3-DEA(70∶30∶ 2∶0.1).Results:The best extraction process was as follows:moisten with ammonia test solution,then leach for 24 hours with 15 times of ether.Conclusion:The optimum extraction methods are rational.

Objective To establish HPLC fingerprint for the identification of Semen Cassiae. MethodsChromatographic fingerprint of Semen Cassiae was investigated by RP-HPLC and the gradient elution mode was applied to chromatographic separation. Data were analysed by fingerprint similarity evaluation software to compare the similarity of the samples. ResultsThe HPLC fingerprint of Semen Cassiae was established preliminarily. ConclusionHPLC Fingerprint method is repeatable and can be used in quality control of Semen Cassiae.

Objective To optimize the extraction technology of alginic acid in Laminaria japonica Aresch. Methods The influence of the amount of solvent, the extraction time and temperature (3 factors) on the alginic acid content and yield was investigated by orthogonal test L9(34), and carried on to inspect the methodology. Result The optimum extraction condition was A1B1C2, that was adding 30 times solvent (1%Na2CO3), whisk extracting for one hour at 60 ℃. Conclusion The temperature is the most important factor. The content of alginic acid decrease obviously at above 60 ℃. The optimum extraction technology is reasonable.