Objective:To determine the contents of calycosin-7-O-?-D-glucoside and formononetin in Astragalus membranaceus(Fisch.) Bge.by high performance liquid chromatography(HPLC).Methods: The HPLC condition was as follows: column: Hypersil ODS 2 column(4.6 mm?250 mm,5 ?m);mobile phase: A was ACN-MeOH(91,V/V),B was H_2O,with gradient elution;flow speed: 1.0 ml/min;detection: 260 nm;temperature of column: room temperature;injection volume: 20 ?l.Astragalus membranaceus(Fisch.) Bge.was extracted with methanol solution twice,each time 20 min.Results: The theoretical plate numbers of calycosin-7-O-?-D-glucoside and formononetin were 50 134 and 25 258,respectively.The calibration curves were linear within the range of(2.022-101.1) ?g/ml for calycosin-7-O-?D-glucoside and(38.04-1522) ?g/ml for formononetin,with their regression function being Y=58 924X-12 352,(r=)(0.999 9) and Y=9 237X-124 447,r=(0.999 9,) respectively.The intra-day and inter-day precisions(RSD) at low,middle and high injection volume were all less than(2.0%.) The limits of detection were(0.202 2) mg/ml for calycosin-7-O-?-D-glucoside and 1.522 mg/ml for formononetin.The recovery rates were 98.34%(RSD=1.33%,n=3) for calycosin-7-O-?-D-glucoside and 98.84%(RSD=0.12%,n=3) for formononetin.The contents of calycosin-7-O-?-D-glucoside and formononetin in 10 different batch of Astragalus membranaceus(Fisch.) Bge.were determined.Conclusion: HPLC is a simple and reliable method for determining the contents of calycosin-7-O-?-D-glucoside and formononetin in Astragalus membranaceus(Fisch.) Bge.

Objective:To re-establish the quality control standard for Weiling granules. Methods:The 4 chief herbs in the prepa-ration, radices paeoniae alba, licorice, rhizoma corydalis and hawthorn were identified by TLC qualitatively. The content of paeoniflor-in in radices paeoniae alba was determined by HPLC. The separation was performed on an Agilent XDB-C18 (250 mm × 4. 6 mm, 5μm)coulme with mobile phase consisting of acetonitrile-0. 1% phosphoric acid solution (10:90). The detection wavelength was 230 nm and the flow rate was 1. 0 ml·min-1 . Results:The spots in TLC were clear without any interference. The linear range for paeoni-florin was 0. 151-1. 212 μg(r=0. 999 9,n=5). The average recovery was 99. 63% and RSD was 2. 01%(n=9). Conclusion:The method is simple and accurate with high reproducibility, which can be used for the quality control of Weiling granules.

Objective:To compare the difference and correlation of HPLC and enzyme-multiplied immunoassay test( EMIT) for the determination of CsA in human whole blood. Methods:A total of 119 clinical samples at different concentrations of CsA were collected and respectively determined by HPLC and EMIT. The difference and correlation of the two determination methods were investigated. Results:There was significant difference in the blood concentrations of CsA determined by HPLC and EMIT(P<0. 05). CsA concen-tration determined by EMIT was 26. 2 ng·ml-1 higher than that determined by HPLC, and 95% CI was (14. 6-37. 7) ng·ml-1 . A satisfactory correlation was achieved between the two methods(r=0. 997 4). Conclusion:There is statistically significant difference in the CsA concentration in whole blood respectively determined by EMIT and HPLC. Attention should be paid to CsA monitoring by E-MIT and HPLC, and relevant adjustment should be carried out.

Objective:To ascertain the optimized ultrasonic condition for extraction of flavone from Radix Astragali by orthogonal design. Methods: The contents of calycosin-7-O-?-D -glucoside and formononetin were taken as the indices and were determined. The ultrasonic time (10 min, 20 min, and 30 min), concentrations of methanol (50%, 75% and 100%) and times of extraction (1, 2, and 3) were analyzed by orthogonal design; the best ultrasonic condition was ascertained and compared with those of soak extraction and Soxhlet extraction. Results: Ultrasonic with 100% methanol twice (20 minutes each time) was the optimized condition for extraction of flavone from Radix Astragali. The efficiency of ultrasonic extraction was better than those of soak extraction and Soxhlet extraction. Conclusion: Compared with other methods, the ultrasonic extraction of flavone from Radix Astragali is efficient, quick and simple.

Objective: To establish the HPLC fingerprints of compound Yinchen granules. Methods: The column was Agilent SB-C18(250 mm×4. 6 mm, 5 μm) and the mobile phase was acetonitrile (A)-0. 2% phosphoric acid solution (B) with gradient elution at a flow rate of 1. 0 ml·min-1. The column temperature was 25℃. The detection wavelength switching technology was used in 180-mi-nute elution time. Results: The HPLC fingerprints of compound Yinchen granules were established. Twenty-two common peaks were confirmed, of which five peaks were identified and 18 peaks were assigned to each crude drug. The overall similarity of the fingerprints of 10 batches of samples was 0. 9 or more when compared with the control map. Conclusion: The fingerprints of compound Yinchen granules can provide reference for the overall quality control of compound Yinchen granules.

Objective To screen potential active anti-cancer components of Brucea javanica.Methods This research has em-ployed comprehensive two dimensional chromatographic technology and cell membrane chromatographic technology simultaneously with mass spectrometry as detector .Results Adenosine and Bruceine B were found to be potentially anti-cancer active .Conclusion This study has combined the advantages of online , high speed and high throughput for the screen of potential active components of traditional Chinese medicine .

Objective To explore specific variables related to cisplatin induced acute kidney injury ,serum metabonomics techniques were applied and simultaneously the value of intervention effects of L-carnitine were appraised .Methods 19 mice were divided into the normal control group ,model group ,and intervention group ,After a three day accommodation period ,the intervention group was given L-carnitine (400 mg/kg ,ip) .Two days later ,cisplatin (20 mg/kg ,ip) was given to the model and intervention groups .The body weight of every mouse in each group was measured daily .Two days after the serum sample of each mice was collected and analyzed by LC-MS ,pattern recognition analysis of metabolomics differences among the groups , and the effectiveness of L-carnitine intervention were evaluated .Results A total of 28 metabolites were identified through ser-um metabolomics analysis .Our data shows that there is a possible mechanism that cisplatin induced AKI was mainly involved in changing phospholipids ,amino acid and fatty acid metabolic pathways and L-carnitine mitigates the damage of acute kidney in-jury induced by cisplatin .Conclusion L-carnitinecan alleviates cisplatin induced acute kidney injury by regulating tryptophan metabolism ,glutamate metabolism ,and energy metabolism .

Knowledge of the evolution of pathogens is of great medical and biological significance to the prevention, diagnosis, and therapy of infectious diseases. In order to understand the origin and evolution of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus), we collected complete genome sequences of all viruses available in GenBank, and made comparative analyses with the SARS-CoV. Genomic signature analysis demonstrates that the coronaviruses all take the TGTT as their richest tetranucleotide except the SARS-CoV. A detailed analysis of the forty-two complete SARS-CoV genome sequences revealed the existence of two distinct genotypes, and showed that these isolates could be classified into four groups. Our manual analysis of the BLASTN results demonstrates that the HE (hemagglutinin-esterase) gene exists in the SARS-CoV, and many mutations made it unfamiliar to us.
Amino Acid Motifs ; Amino Acid Substitution ; Base Composition ; Codon ; genetics ; Computational Biology ; DNA Mutational Analysis ; Evolution, Molecular ; Gene Transfer, Horizontal ; Genetic Variation ; Genome, Viral ; Phylogeny ; SARS Virus ; genetics

Annotation of the genome sequence of the SARS-CoV (severe acute respiratory syndrome-associated coronavirus) is indispensable to understand its evolution and pathogenesis. We have performed a full annotation of the SARS-CoV genome sequences by using annotation programs publicly available or developed by ourselves. Totally, 21 open reading frames (ORFs) of genes or putative uncharacterized proteins (PUPs) were predicted. Seven PUPs had not been reported previously, and two of them were predicted to contain transmembrane regions. Eight ORFs partially overlapped with or embedded into those of known genes, revealing that the SARS-CoV genome is a small and compact one with overlapped coding regions. The most striking discovery is that an ORF locates on the minus strand. We have also annotated non-coding regions and identified the transcription regulating sequences (TRS) in the intergenic regions. The analysis of TRS supports the minus strand extending transcription mechanism of coronavirus. The SNP analysis of different isolates reveals that mutations of the sequences do not affect the prediction results of ORFs.
Amino Acid Substitution ; Base Composition ; Base Sequence ; Computational Biology ; methods ; Genome, Viral ; Isoelectric Point ; Models, Genetic ; Molecular Sequence Data ; Molecular Weight ; Open Reading Frames ; SARS Virus ; genetics ; Sequence Analysis ; Transcription, Genetic

The R (replicase) protein is the uniquely defined non-structural protein (NSP) responsible for RNA replication, mutation rate or fidelity, regulation of transcription in coronaviruses and many other ssRNA viruses. Based on our complete genome sequences of four isolates (BJ01-BJ04) of SARS-CoV from Beijing, China, we analyzed the structure and predicted functions of the R protein in comparison with 13 other isolates of SARS-CoV and 6 other coronaviruses. The entire ORF (open-reading frame) encodes for two major enzyme activities, RNA-dependent RNA polymerase (RdRp) and proteinase activities. The R polyprotein undergoes a complex proteolytic process to produce 15 function-related peptides. A hydrophobic domain (HOD) and a hydrophilic domain (HID) are newly identified within NSP1. The substitution rate of the R protein is close to the average of the SARS-CoV genome. The functional domains in all NSPs of the R protein give different phylogenetic results that suggest their different mutation rate under selective pressure. Eleven highly conserved regions in RdRp and twelve cleavage sites by 3CLP (chymotrypsin-like protein) have been identified as potential drug targets. Findings suggest that it is possible to obtain information about the phylogeny of SARS-CoV, as well as potential tools for drug design, genotyping and diagnostics of SARS.
Amino Acid Sequence ; Base Composition ; Base Sequence ; Cluster Analysis ; Computational Biology ; Conserved Sequence ; genetics ; Evolution, Molecular ; Gene Components ; Genome, Viral ; Molecular Sequence Data ; Mutation ; genetics ; Phylogeny ; Protein Structure, Tertiary ; RNA Replicase ; genetics ; SARS Virus ; genetics ; Sequence Analysis, DNA