2.Preliminary Study of Bio-transformation of Phytosterol by HPLC-MS
Wei SHEN ; Wei-Hong LIAO ; Zhi-Ming RAO ; Hui-Ying FANG ; Jian ZHUGE ;
Microbiology 2008;0(10):-
The method for analysis and determination the cleavage of soybean sterol, in which the soybean sterol was degraded and the products androst-1,4-diene-,17-dione (ADD) and androst-4-ene-3,17-dion (AD) were developed by Liquid Chromatography-mass Spectrometry. The HPLC conditions adopted were: a All- tima ODS-2 column (250 mm?4.6 mm, 5 ?m), a mobile phase consisted of menthanol-water (70:30), a flow rate of 1.0 mL/min, a room column temperature. and the detective wavelength was 244 nm.The ZMD Micromass electrospray ionization (ESI)-mass spectrometer was employed. In such conditions the corre- sponding HPLC chromatogram and MS spectrum were obtained. The method has a linear ranger of 0.01 mg/mL ~ 0.09 mg/mL, R2 =0.9999, the recoveries of ADD and AD were 102.6% and 105.90%, the RSD of ADD and AD were 3.02%, 3.5% and 3.08%, 3.24%. This method showed high sensitivity, accuracyand easy to perform. It is suitable to analysis the process cleavage of soybean sterol as well as quality control of product.
3.Studies on chemical constituents in Flos Sophorae Carbonisatus.
Rao-rao LI ; Si-tong YUAN ; Zhi-hong LI ; Hai-lin QIN
China Journal of Chinese Materia Medica 2005;30(16):1255-1257
OBJECTIVETo investigate the chemical constituents in Flos Sophorae Carbonisatus.
METHODSilica gel column chromatography was used to separate and purify the chemical constituents. The structures were elucidated by spectral analysis.
RESULTSix compounds were isolated from Flos Sophorae Carbionisatus, and their structures were elucidated as maltol (1), 3-hydroxypyridine (2), malto-3-O-[6'-O-(4"-hydroxy-tans-cinnamoyl)-beta-D-glucopyranoside (3), 3-O-[beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl] sophoradiol ethyl ester (4), 3-O-[beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl] sophoradiol methyl ester (5), rutin (6).
CONCLUSION4 is a new compound, and 1,2,3,5 were first reported from Flos Sophorae Carbonisatus.
Coumaric Acids ; chemistry ; isolation & purification ; Flowers ; chemistry ; Glucosides ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Pyridines ; chemistry ; isolation & purification ; Pyrones ; chemistry ; isolation & purification ; Saponins ; chemistry ; isolation & purification ; Sophora ; chemistry
4.Effect on the Synthesis of 1,3 propanediol of Organic acids in the Two-stage Fermentation
Ji-Hong LU ; Ge-Bin ZHU ; Wei SHEN ; Zhi-Ming RAO ; Hui-Ying FANG ; Ge-Jian ZHU ;
China Biotechnology 2006;0(05):-
Effect of organic acids on the synthesis of 1,3 propanediol was studied.The adsorption of organic acids from glycerol fermentation liquor by ion-exchange resins was investigated.The results showed that organic acid and 1,3 propanediol production was in negative relationship.The static adsorption showed that ion-exchange resin 005 had the best adsorption abilities of the organic acids in the glycerol fermentation liquor.It was showed that the yield of 1,3propanediol increased by 166% after the extraction of organic acids from glycerol fermentation liquor and the convertion rate increased by 34%.
5.Study on methods for quality control of recombinant human neu epitope peptide 12.
Hong JI ; Jun-zhi WANG ; Chun-ming RAO ; Yi ZHANG ; Min WANG
Acta Pharmaceutica Sinica 2004;39(5):359-362
AIMTo establish methods and requirements for the quality control of recombinant human neu epitope peptide 12.
METHODSBiological activity of recombinant human neu epitope peptide 12 was evaluated in FVB/N transgenic mice (TgN MMTV neu 202 Mul, Jackson Lab., USA) administered with samples. The percentage of antibody-positive mice detected by ELISA was used in the biological activity evaluation. The peptide map was performed by peptic digestion. The antigen content was determined by SEC-HPLC.
RESULTS AND CONCLUSIONThe quality control methods, such as biological activity, peptide map, antigen content, and the requirements for the quality control of recombinant human neu epitope peptide 12 were established. The established methods and requirements were already used for the quality control of recombinant human neu epitope peptide 12 products.
Animals ; Antibodies, Monoclonal ; analysis ; Biotechnology ; methods ; Epitopes ; analysis ; chemistry ; immunology ; Female ; Genes, erbB-2 ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Transgenic ; Peptide Fragments ; chemistry ; immunology ; Peptide Mapping ; Quality Control ; Receptor, ErbB-2 ; analysis ; chemistry ; immunology ; Recombinant Fusion Proteins ; analysis ; chemistry ; immunology ; Technology, Pharmaceutical ; standards
6.Study of pharmacokinetics of digoxin in ovariectomized rats model.
Yong-wen JIN ; Hong-yan QIN ; Zhi RAO ; Guo-qiang ZHANG ; Yan Rong MA ; Yu-Hui WEI ; Xin-an WU
Acta Pharmaceutica Sinica 2015;50(12):1603-1606
This study aims to investigate the change of plasma concentration of digoxin (DIG) in rats with ovariectomy. Twelve female SD rats were randomly assigned into ovariectomized group and sham group (n = 6). All rats plasma was collected after a single dose of 2 mg x kg(-1) DIG administrated orally, serum DIG concentration was determined by LC-MS/MS. The level of P-gp in the intestinal was analyzed by Western blotting. Pharmacokinetic calculations were performed on each individual using DAS 2.0 practical pharmacokinetic software. Compared with the sham group, C(max) of ovariectomized group decreased significantly (P < 0.01). There was no significant difference of AUC(0-t), and the level of P-gp was elevated in ovariectomized group. It was found that C(max) of DIG was significantly reduced after ovariectomy, and the change was associated with the decreased level of estrogen, which contributes to the increased level of P-gp.
ATP-Binding Cassette, Sub-Family B, Member 1
;
metabolism
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Animals
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Blotting, Western
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Chromatography, Liquid
;
Digoxin
;
blood
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pharmacokinetics
;
Disease Models, Animal
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Estrogens
;
blood
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Female
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Ovariectomy
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Rats
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Rats, Sprague-Dawley
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Tandem Mass Spectrometry
7.DNA quantification of blood samples pre-treated with pyramidon.
Chuan-Hong ZHU ; Dao-Li ZHENG ; Rao-Zhi NI ; Hai-Sheng WANG ; Ping NING ; Hui FANG ; Yan LIU
Journal of Forensic Medicine 2014;30(3):188-190
OBJECTIVE:
To study DNA quantification and STR typing of samples pre-treated with pyramidon.
METHODS:
The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology.
RESULTS:
In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples.
CONCLUSION
Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.
Aminopyrine/pharmacology*
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Blood Stains
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DNA/isolation & purification*
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DNA Fingerprinting
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Forensic Medicine
;
Humans
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Polymerase Chain Reaction
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Reproducibility of Results
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Specimen Handling
8.Selection of sentinel sites for death surveillance,using cluster or unequal probability sampling
Heng-Li LIAN ; Yong-Yong XU ; Ling-Xia GUO ; Zhi-Jun TAN ; Dan-Hong LIU ; Ke-Qin RAO
Chinese Journal of Epidemiology 2010;31(4):459-461
To compare the sampling errors from cluster or unequal probability sampling designs and to adopt the unequal probability sampling method to be used for death surveillance.Taking 107 areas from the county level in Shaanxi province as the sampling frame,a set of samples are drawn by equal probability cluster sampling and unequal probability designs methodologies.Sampling error and effect of each design are estimated according to their complex sample plans.Both the sampling errors depend on the sampling plan and the errors of equal probability in stratified cluster sampling appeares to be less than simple cluster sampling.The design effects of unequal probability stratified cluster sampling,such as πPS design,are slightly lower than those of equal probability stratified cluster sampling,but the unequal probability stratified cluster sampling can cover a wider scope of monitoring population.Conclusions:Results from the analysis of sampling data can not be conducted without consideration of the sampling plan when the sampling frame is finite and a given sampling plan and parameters,such as sampling proportion and population weights,are assigned in advance.Unequal probability cluster sampling designs seems to be more appropriate in selecting the national death surveillance sites since more available monitoring data can be obtained and having more weight in estimating the mortality for the whole province or the municipality to be selected.
9.Quality control methods for recombinant human endostatin.
Yong-Hong LI ; Jun-zhi WANG ; Chun-mei HAN ; Yi ZHANG ; Chun-ming RAO
Acta Pharmaceutica Sinica 2002;37(10):807-811
AIMTo establish the quality control methods for recombinant human endostatin.
METHODSBiological activity was determined by endothelial cell migration assays. Peptide mapping was tested by trypsin digestion and RP-HPLC. Purity was determined by non-reduced SDS-PAGE and RP-HPLC. Other tests including molecular weight, isoelectrical point, etc. were done according to the National Requirements for Biological Products (2000).
RESULTSThe method of bioassay was established and used for determining activity of endostatin. Specific activity of the three batchs of drug substance was 1.45 x 10(6), 1.57 x 10(6) and 2.73 x 10(6) u.mg-1 proteins. Peptide mappings of the three batches of drug substance were completely identified. Both purity results of the products tested by SDS-PAGE and RP-HPLC were more than 99%.
CONCLUSIONThe established methods can effectively control the quality of recombinant human endostatin.
Cell Movement ; drug effects ; Cells, Cultured ; Endostatins ; pharmacology ; Endothelium, Vascular ; cytology ; Humans ; Quality Control ; Recombinant Proteins ; pharmacology ; Technology, Pharmaceutical ; methods
10.Expression of pig7 in acute leukemia and its clinical significance.
Dong-hai WANG ; Yang WANG ; Min WANG ; Hang LIU ; Zhi-fang XU ; Qing RAO ; Ji-hong MENG ; Jian-xiang WANG
Chinese Journal of Hematology 2007;28(8):532-536
OBJECTIVETo investigate pig7 expression level in acute leukemia (AL) and its clinical significance and explore the possible mechanisms for pig7 silence in terms of methylation control.
METHODSExpression levels of pig7 mRNA in bone marrow samples from 138 patients with de novo AL and 21 normal controls and in 6 leukemic cell lines were detected by quantitative real-time reverse transcription PCR (RT-PCR). Differentiation induction effect by all-trans retinoic acid (ATRA) and concomitant change in pig7 expression were also monitored in NB4 cells. Endonuclease analysis was employed to determined the identity of pig7 transcript present in AL samples. Methylation specific PCR (MSP) was used to elucidate if hypermethylation was responsible for pig7 silence in AL.
RESULTSCompared with that in normal control, pig7 expression was markedly decreased (0.62 vs 18.30, median, P < 0.01) in AL patients on progression (at diagnosis, relapse or refractory). No significant difference was observed between acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL). AL at diagnosis had a higher pig7 level than those with relapsed or refractory disease (1.43 vs 0.16, median, P < 0.05). The complete remission (CR) rate after chemotherapy was found to be significantly correlated with pig7 expression levels (P < 0.05). Differentiated NB4 cells showed an increased level of pig7 expression (from 1.61 +/- 0.72 to 44.75 +/- 3.93, P < 0.01). Only one form of pig7 transcripts i.e., Small integral membrane protein of late endosome (SIMPLE), was detected in AL patients. Hypermethylation of pig7 promoter was identified in K562 and HL-60 cells, in contrast to non-methylation predominant in U937 cells.
CONCLUSIONAberrant down-regulation of pig7 provides novel insights into leukemogenesis and therapy response prediction in AL.
Acute Disease ; Cell Differentiation ; Cell Line, Tumor ; DNA Methylation ; Gene Expression Regulation, Leukemic ; Humans ; Leukemia ; genetics ; Nuclear Proteins ; genetics ; Promoter Regions, Genetic ; RNA, Messenger ; genetics ; Transcription Factors ; genetics ; Tretinoin ; pharmacology