Eight phenolic compounds were isolated from Rhododendron phaeochrysum var. agglutinatum and their sructures were identified as phaeochrysin (1), (2R)-4-(3',4'-dihydroxyphenyl) -2-butanol (2), (-) -rhododendrol (3), rhododendrin (4), (+) -isolariciresinol (5), (-) -lyoniresinol (6), lyoniresinol-9'-O-β-D-xylopyranoside (7), and dihydrodehydrodiconiferyl-3a-O-α-L-rhamnopyranoside (8). Compound 1 is new, and compounds 2, 5-8 were isolated from this plant for the first time.
Drugs, Chinese Herbal ; chemistry ; Mass Spectrometry ; Molecular Structure ; Phenols ; chemistry ; Rhododendron ; chemistry

Human ITP clinical trials have been performed based on observations of clinical uses of drugs, other than on mechanism studies in preclinical models, many studies were stopped because of unexpected complications. So there was need to develop ideal animal model of ITP to improve the understanding of the pathophysiology and the mechanisms of action of therapeutics. In this review, the history of animal models of ITP was retrospected, two modeling methods inducing passive and active models, especially transgenic rice model which can more accurately represent human disease pathophysiology, and their application in study of ITP were summarized. An animal model for human immune thrombocytopenia allows detailed studies of the mechanism, kinetics, and therapy of human immune thrombocytopenia.
Animals ; Disease Models, Animal ; Mice ; Mice, Transgenic ; Purpura, Thrombocytopenic, Idiopathic ; Rats

OBJECTIVETo establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen III N-terminal peptide (P III NP) in serum.

METHODSA sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P III NP as the catalytic enzyme and the luminol as the luminescence reagent. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. The CLEIA was compared with imported ELISA kits, by detecting clinical serum.

RESULTSThe linear range was 0.8-85 ng/ml. The detection limit was 0.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 96.2%, 91.2% and 101.1%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6%. The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA.

CONCLUSIONEstablished CLEIA for quantity determination of serum P III NP has high accuracy, sensitivity and repeatability.

Adult ; Aged ; Antibodies, Monoclonal ; analysis ; Enzyme-Linked Immunosorbent Assay ; instrumentation ; methods ; Female ; Humans ; Liver Cirrhosis ; blood ; diagnosis ; Luminescence ; Luminescent Measurements ; methods ; Male ; Middle Aged ; Peptide Fragments ; blood ; chemistry ; Procollagen ; blood ; chemistry ; Sensitivity and Specificity

OBJECTIVETo establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum.

METHODSA sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on.

RESULTSThe linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%.

CONCLUSIONEstablished ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.

Antibodies, Monoclonal ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Membrane Proteins ; blood ; Sensitivity and Specificity

OBJECTIVETo establish a quick method for evaluation of the antioxidant activities based on the correlation analysis of lignans content and antioxidant activities of Schisandra chinensis fruits.

METHODThe content of five lignans components in 37 batches of S. chinensis fruits from different regions of Jilin province were measured by HPLC. Simultaneously, the antioxidant activities of the above samples were detected, such as lipid peroxidation inhibition activity in liver (LPIL), kidney (LPIK) and brain (LPIB) and the clearance rate of DPPH (CRD). Bivariate correlation analysis and stepwise regression analysis were carried out by the software of SPSS for windows 11.5.

RESULTThe results of bivariate correlation analysis showed that deoxyschizandrin was negative correlation (P<0.01) to the activity of LPIL, LPIB, CRD. Schisandrin was positive correlation (P<0.01) to the activity of LPIL, LPIB, CRD. Schisandrol B was also positive correlation (P<0.05 or P<0.01) to the above four kinds of antioxidant activity. The results of stepwise regression analysis were mostly consistent with the bivariate correlation analysis results. For the other 10 batches of samples, the simulated antioxidant activities according to the regression equation calculated was consistent with the measured activities.

CONCLUSIONBy using the bivariate correlation analysis and linear stepwise regression analysis, the bioactive components related to the antioxidant activity of S. chinensis fruits were found. Meanwhile, the antioxidant activity of samples will be inferred according to the content of Schisandra lignans.

Antioxidants ; chemistry ; Fruit ; chemistry ; Lignans ; chemistry ; Schisandra ; chemistry

Biomarkers, Tumor ; Bronchi ; metabolism ; DNA Methylation ; Genes, p16 ; Humans ; Lung Neoplasms ; diagnosis ; genetics ; Mucous Membrane ; metabolism ; Promoter Regions, Genetic ; Sensitivity and Specificity

OBJECTIVETo prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein.

METHODSTIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs.

RESULTSThe prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein.

CONCLUSIONThe TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.

Animals ; Antibodies, Monoclonal ; immunology ; Cloning, Molecular ; Humans ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology ; isolation & purification ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; immunology

OBJECTIVETo establish a purificatory method of alpha-fetoprotein variant (AFP-L3) based on microspincolumn with lens culinaris agglutinin (LCA).

METHODSLCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectrophoresis.

RESULTS8 of 10 cases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment.

CONCLUSIONSuccessfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.

Adsorption ; Chromatography, Affinity ; methods ; Immunoelectrophoresis ; Lens Plant ; Plant Lectins ; chemistry ; Reproducibility of Results ; alpha-Fetoproteins ; chemistry ; isolation & purification

Objective:To detect IL-35 in mononuclear cells of peripheral blood and lung tissue with BCG infected mouse,and to analyze its effect on the tuberculosis immune mechanism and pathology.Methods: Mycobacterium tuberculosis infecting animal model was made by BCG injection in wild type C57BL/6 mouse.Then,mice were sacrificed in different time points.The peripheral blood and lung tissue were isolated.The bacterial load and histopathological analysis in lung tissue were performed,and the expression of IL-35 subunits P35 and EBI3 in monocytes were detected by flow cytometry.Furthermore,the correlation of IL-35-expressing monocytes with the load of bacterium was analyzed.Results:The expression of P35 and EBI3 in monocytes in peripheral blood and lung tissue of BCG-infected mice was significantly higher than that of the control group,and the peripheral blood of the experimental group was significantly increased at 4 weeks after infection.The expression of EBI3 in lung tissue diaplayed significant rising trend.The correlation analysis showed that the P35 expression was positively correlated with the expression of EBI3 in monocyte,and the IL-35-ex-pressing monocyte in lung tissue was negatively correlated with the load of bacteria.The IL-35-expressing monocyte in peripheral blood and lungs increased significantly at 2-week and 4-week,while it increased in lung tissue at 8 week,significantly.Conclusion: BCG-infects mouse monocytes with high expression of IL-35,it may participate in anti-tuberculosis immune regulation.

BACKGROUNDRecent studies suggest that circulating DNA may be a potential tumor marker for lung cancer, but most of these studies are conducted between healthy controls and lung cancer patients, with few or no benign lung disease patients included. The objective of this study was to evaluate the performance of plasma DNA quantification in discriminating lung cancer from the healthy and benign lung disease.

METHODSPlasma DNA was extracted with a QIAamp DNA Blood Midi kit and quantified by a PicoGreen dsDNA quantitation kit in 44 healthy individuals, 36 benign lung disease patients and 67 lung cancer patients. Discrimination power was evaluated by the receiver operating characteristic curve.

RESULTSPlasma DNA values were significantly increased in lung cancer patients, especially in those with metastases, and in benign lung disease patients compared with that in the healthy individuals (P < 0.001, respectively). The values in lung cancer patients were significantly increased compared with that in the benign lung disease patients (P < 0.001). The area under the curve was 0.96 [95% confidence interval (CI) 0.92 - 0.99] for the healthy versus lung cancer, 0.73 (95% CI 0.64 - 0.83) for lung cancer versus benign lung disease, and 0.86 (95% CI 0.80 - 0.91) for lung cancer versus the healthy and benign lung disease.

CONCLUSIONSPlasma DNA quantification has a strong power to discriminate lung cancer from the healthy and from the healthy and benign lung disease, less power to discriminate lung cancer from benign lung disease. Plasma DNA quantification may be useful as a screening tool for lung cancer.

DNA ; blood ; Humans ; Lung Neoplasms ; blood ; diagnosis ; pathology ; Neoplasm Staging