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1.
Zhongguo Zhong Yao Za Zhi ; 39(12): 2295-9, 2014 Jun.
Article in Chinese | MEDLINE | ID: mdl-25244762

ABSTRACT

The establishment of high specificity and sensitivity method of small molecule monoclonal antibody-based immunoassay has a great importance in the study of small molecule compounds in Chinese medicine, wherein synthesis of small molecule artificial antigen is a critical step in the preparation of small molecule antibodies. Oxidation method using sodium iodide was used to synthesize immunogenic antigen (FRn-BSA) and coating antigen (FRn-OVA) of forsythin. UV spectroscopy and thin layer chromatography showed that forsythin was successfully conjugated with BSA and OVA. After immuned FRn-BSA, the mice could specifically produce anti-forsythin antibodies with titer up to 1:8 000, and the linear range was from 1 mg x L(-1) to 100 mg x L(-1). In this paper, the artificial antigen of forsythin was successfully synthesized, which can be applied for preparation of monoclonal antibodies and establishment of appropriate immune method.


Subject(s)
Antigens/immunology , Bridged Bicyclo Compounds, Heterocyclic/immunology , Drugs, Chinese Herbal/chemistry , Furans/immunology , Animals , Antibodies/immunology , Antigens/chemistry , Bridged Bicyclo Compounds, Heterocyclic/chemistry , Furans/chemistry , Male , Mice , Mice, Inbred BALB C
2.
Biol Pharm Bull ; 37(9): 1525-33, 2014.
Article in English | MEDLINE | ID: mdl-25177035

ABSTRACT

Geniposide, Geniposide, the main active component in extracts of Gardenia jasminoides ELLIS., is one of the main components of Huanglian-Jiedu-Tang (HJT). This study aimed to validate an indirect competitive enzyme-linked immunosorbent assay (icELISA) based on monoclonal antibodies (mAb) against geniposide (anti-geniposide mAb), which was developed by our lab, and apply the assay to study the pharmacokinetics of geniposide in HJT in mice. Blood samples were drawn from mice at predetermined time points after oral administration of HJT in three dosages. A linear correlation was obtained for geniposide concentrations in the range from 1.17 to 37.50 µg/mL. The intra-day and inter-day precision values of the icELISA method were well within the recommended range (≤10%). The recovery rates ranged from 99.74 to 102.40%. Stability studies showed that geniposide sample solutions were intact for 12 h. The Tmax and mean residence time (MRT) of geniposide of the three groups were consistent with previous data. The results suggest that a reliable and effective method was established and could be applied to the study of the pharmacokinetics of geniposide in HJT.


Subject(s)
Drugs, Chinese Herbal/pharmacokinetics , Iridoids/blood , Administration, Oral , Animals , Antibodies, Monoclonal/immunology , Chromatography, High Pressure Liquid , Drugs, Chinese Herbal/chemistry , Enzyme-Linked Immunosorbent Assay , Iridoids/analysis , Iridoids/immunology , Male , Mice, Inbred BALB C , Reproducibility of Results
3.
Zhong Xi Yi Jie He Xue Bao ; 5(3): 322-7, 2007 May.
Article in Chinese | MEDLINE | ID: mdl-17498495

ABSTRACT

OBJECTIVE: To investigate the effects of Ganoderma spores on mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension. METHODS: Nitric oxide synthase (NOS) inhibitor Nw-nitro-L-arginine methylester (L-NAME) was intraperitoneally injected into pregnant rats to induce gestational hypertension, and Ganoderma spores were administered orally. The effects of Ganoderma spores on levels of mitochondria-related molecular substances in hippocampus of young rats birthed by the rats with gestational hypertension were evaluated with immunoradiometric assay of cAMP, RT-PCR analysis of related genes, and detection of enzyme activity. RESULTS: In hippocampus of the new-born rats birthed by rats with gestational hypertension, the cAMP level, mitochondrial DNA (mtDNA) level and adenosine triphosphatase (ATPase) activity were decreased, and the expression level of peroxisome proliferator activated receptor gamma coactivator 1 alpha (pgc1 alpha) was unchanged compared to the normal control group. The cAMP level, mtDNA level, ATPase activity and pgc1 alpha expression level in hippocampus of 30-day post-natal rats were lower than those of the rats in normal control group. After oral administration of Ganoderma spores, the cAMP and mtDNA levels in hippocampus of the new-born rats and 30-day post-natal rats recovered almost to the levels of normal control rats, and the ATPase activity and pgc1 alpha expression level were also increased significantly. CONCLUSION: Ganoderma spores may regulate the levels of mitochondria-related molecular substances in hippocampus of young rats birthed by rats with gestational hypertension.


Subject(s)
DNA, Mitochondrial/metabolism , Ganoderma/physiology , Hippocampus/metabolism , Hypertension, Pregnancy-Induced/physiopathology , Mitochondrial Proteins/metabolism , Animals , Animals, Newborn , Cyclic AMP/metabolism , DNA, Mitochondrial/genetics , Female , Ganoderma/chemistry , Hypertension, Pregnancy-Induced/chemically induced , Male , Mitochondrial Proteins/genetics , NG-Nitroarginine Methyl Ester , Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha , Pregnancy , RNA-Binding Proteins , Rats , Rats, Sprague-Dawley , Reverse Transcriptase Polymerase Chain Reaction , Spores/chemistry , Spores/physiology , Transcription Factors/metabolism
4.
Zhong Xi Yi Jie He Xue Bao ; 4(3): 298-302, 2006 May.
Article in Chinese | MEDLINE | ID: mdl-16696920

ABSTRACT

OBJECTIVE: To detect some proteins associated with the effect of ganoderma lucidium spores (GASP) on promoting the survival and axon regeneration of injured spinal motor neurons in rats. METHODS: The rats were divided into normal control group, untreated group and GASP-treated group, and the rats in the last two groups received ventral root avulsion. GASP preparation was fed to the rats in the GASP-treated group for 14 days. The gray matter tissues of the lumbar spinal were sampled from rats in each group after 14 days following ventral root avulsion, and the extracted proteins from these tissues were detected by using 2-dimensional electrophoresis. Matrix assisted laser desorption/ionization time-of-flight mass spectroscopy (MALDI-TOF MS) was utilized to identify the differentially expressed proteins among these three groups. RESULTS: There were six kinds of proteins differentially expressed among the three groups, which were collapsin response mediator protein 2 (CRMP-2), F-actin capping protein beta subunit (FCP-beta), isocitrate dehydrogenase [NAD] subunit beta (IDH-beta), ATPase, glutamate oxaloacetate transaminase-1 (GOT1) and M2 pyruvate kinase (M2-PK). The expression levels of CRMP-2, IDH-beta, ATPase and GOT1 were higher in the GASP-treated group than those in the untreated group, while the expression levels of FCP-beta and M2-PK were lower than those in the untreated group. CONCLUSION: GASP maybe promotes the survival and axon regeneration of injured spinal motor neurons in rats by virtue of up- or down-regulating the expression levels of the proteins mentioned above.


Subject(s)
Motor Neurons/physiology , Nerve Regeneration/drug effects , Phytotherapy , Reishi , Spinal Cord Injuries/drug therapy , Actin Capping Proteins/biosynthesis , Actin Capping Proteins/genetics , Animals , Axons/physiology , Cell Survival , Drugs, Chinese Herbal/pharmacology , Drugs, Chinese Herbal/therapeutic use , Intercellular Signaling Peptides and Proteins , Isocitrate Dehydrogenase/biosynthesis , Isocitrate Dehydrogenase/genetics , Nerve Tissue Proteins/biosynthesis , Nerve Tissue Proteins/genetics , Proteomics , Random Allocation , Rats , Rats, Sprague-Dawley , Spinal Cord Injuries/physiopathology , Spores, Fungal
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