ABSTRACT
A boy, aged 6 years and 11 months, was admitted due to nephrotic syndrome for 2 years, cough for 1 month, and shortness of breath for 15 days. The boy had a history of treatment with hormone and immunosuppressant. Chest CT after the onset of cough and shortness of breath showed diffuse ground-glass opacities in both lungs. Serum (1, 3)-beta-D glucan was tested positive, and the nucleic acid of cytomegalovirus was detected in respiratory secretions. After the anti-fungal and anti-viral treatment, the child improved temporarily but worsened again within a short period of time.
Subject(s)
Child , Humans , Male , Cough/etiology , Cytomegalovirus Infections/therapy , Dyspnea/etiology , Extracorporeal Membrane Oxygenation , Nephrotic Syndrome/complications , Pneumonia, Pneumocystis/therapy , Respiratory Distress Syndrome, Newborn/therapyABSTRACT
OBJECTIVES@#To set up ELISA for detection of atrazine with high precision.@*METHODS@#The reaction condition of indirect-ELISA was optimized, including atrazine-ovalbumin(AT-OVA) concentration and primary antibody concentration, organic solvent, goat anti-rat immunoglobin G-horseradish peroxidase(IgG-HRP) concentration. The actual samples were detected by the ELISA method established in our laboratory. Then the ELISA method was compared with the HPLC.@*RESULTS@#The specification curve of indirect-ELISA was set up after optimization. The relation coefficient R=0.9958. The limit of detection (LOD) was 1.972 ng/ml. The percent recovery of the actual samples was in range of 80%~120%. The ELISA detection sensitivity was higher than the HPLC in the range of 0 ng/ml~6 ng/ml atrazine.@*CONCLUSIONS@#The ELISA to detect atrazine has good specificity and high precision. And it can be applied in testing real atrazine samples replacing of the large-scale instrument.
Subject(s)
Animals , Atrazine , Chromatography, High Pressure Liquid , Enzyme-Linked Immunosorbent Assay , Sensitivity and SpecificityABSTRACT
During the screening of a traditional Chinese folk herb library against HepG2 and Hep3B cell lines, the EtOAc extract from the Tibetan medicine, Caragana tibetica (CT-EtOAc) exhibited potential anti-hepatocellular carcinoma (anti-HCC) activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-EtOAc by MS-directed purification method. CT-EtOAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column (150 mm × 10 mm i.d., 5 μm) with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-EtOAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone (CT-1) and 3-hydroxy-8, 9-dimethoxypterocarpan (CT-2) relevant to anti-HCC effects for the EtOAc extract of C. tibetica rapidly and unambiguously by this protocol.
ABSTRACT
During the screening of a traditional Chinese folk herb library against HepG2 and Hep3B cell lines, the EtOAc extract from the Tibetan medicine, Caragana tibetica (CT-EtOAc) exhibited potential anti-hepatocellular carcinoma (anti-HCC) activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-EtOAc by MS-directed purification method. CT-EtOAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column (150 mm × 10 mm i.d., 5 μm) with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-EtOAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone (CT-1) and 3-hydroxy-8, 9-dimethoxypterocarpan (CT-2) relevant to anti-HCC effects for the EtOAc extract of C. tibetica rapidly and unambiguously by this protocol.
Subject(s)
Humans , Acetates , Pharmacology , Antineoplastic Agents , Chemistry , Pharmacology , Caragana , Chemistry , Carcinoma, Hepatocellular , Drug Therapy , Cell Line, Tumor , Chalcones , Pharmacology , Chromatography, High Pressure Liquid , Hep G2 Cells , Liver Neoplasms , Drug Therapy , Medicine, Tibetan Traditional , Plant Extracts , Chemistry , Pharmacology , Plant Roots , ChemistryABSTRACT
<p><b>OBJECTIVE</b>To identify the self-preparation monoclonal antibody which target to clenbuterol, and set up the standard curve to clenbuterol (CL) detection.</p><p><b>METHODS</b>The affinity constants and activity of the monoclonal antibody which target to CL were determined by ELISA. ELISA was also used to confirm whether the monoclonal antibody had any across-reaction with BSA and CL analogues. The rat ascites which contains the monoclonal antibody target to CL was purified by (NH4)2SO4 salt-out method and further by affinity column. At last, the CL detection standard curve which based on indirect competition ELISA was established.</p><p><b>RESULTS</b>The ELISA experiment showed that the antibody titer was 10(6) and the monoclonal antibody affinity constants was 2.90 x 10(10) L/mol. The result of the indirect competition ELISA confirmed that the monoclonal antibody had no cross-reaction with BSA and a few kind of CL analogue. CL detection standard curve based on indirect competition ELISA was established, which R2 was 0.9812, and the lowest detectable limit was 1.0 ng/ml.</p><p><b>CONCLUSION</b>The standard curve based on indirectly competitioning ELISA was established. The self-preparation monoclonal antibody which target to CL has high affinity and high specific to CL, which had established the foundation to the advanced development of the CL immune test paper and CL ELISA kit.</p>
Subject(s)
Animals , Rats , Antibodies, Monoclonal , Chemistry , Antibody Affinity , Clenbuterol , Allergy and Immunology , Cross Reactions , Enzyme-Linked Immunosorbent Assay , Limit of DetectionABSTRACT
<p><b>AIM</b>To obtain Clenbuterol monoclonal antibodies.</p><p><b>METHODS</b>Clenbuterol complete antigen was prepared with diazotization method. BALB/c mice was immunized with subtractive immunization, Clenbuterol monoclonal antibody was prepared with rule hybridoma technique.</p><p><b>RESULTS</b>The mice obtained tolerance to BSA by subtractive immunization. The rate of the hybridoma cell with positive reaction which had obtained was 8.2%, and the specific clenbuterol monoclonal antibody was obtained at last.</p><p><b>CONCLUSION</b>Monoclonal antibodies to micromolecule contaminant be prepared by subtractive immunization, could decrease the workload in the bolting of monoclonal antibodies, and increase the chance to obtain the antibody of expected.</p>
Subject(s)
Animals , Female , Male , Mice , Antibodies, Monoclonal , Allergy and Immunology , Clenbuterol , Allergy and Immunology , Hybridomas , Metabolism , Immunization , Methods , Mice, Inbred BALB C , Serum Albumin, Bovine , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To observe the clinical therapeutic effect of Qingre Cuochuang tablet (QCT) in treating female delayed acne vulgaris (FDAV, with patients age more than 25 years old), to evaluate objectively the sexual hormone in patients and to assess the effect of QCT on sexual hormone.</p><p><b>METHODS</b>Sixty FDAV patients were randomly divided into the treated group (n = 40) and the control group (n = 20), they were treated with QCT and western medicine (including antisterone, tetracycline and metronidazole) respectively. Besides, 10 healthy female subjects aged > or = 25 years were selected as normal control. Serum levels of testosterone (T) and estradiol (E2) in all patients and healthy subjects as well as the clinical therapeutic effect of the treatments were observed and compared.</p><p><b>RESULTS</b>The total effective rate in the treated group and the control group was 92.5% and 90.0% respectively, comparison between them showed insignificant difference. Serum level of T in the patients before treatment were higher than that in healthy subjects (P < 0.01), and showed no difference between the treated group and the control group. After treatment, it lowered significantly in the treated group (P < 0.01), but unchanged in the control group, E2 level showed no significant change in both groups before and after treatment.</p><p><b>CONCLUSION</b>QCT has definite clinical effect in treating FDAV, it could lower the serum level of T and with few adverse reaction.</p>