OBJECTIVETo study the alkaloid constituents of Corydalis adunca.

METHODThe constituents were isolated on silica gel column and their structures were elucidated by IR, NMR, MS data.

RESULTEight alkaloid compounds were isolated from alcohol extracts of the herb of C. adunca, and identified as dihydrosanguinarine (I), tetrahydrocolumbamine (II), 1,2,3,4-tetrahydro-7-methoxy-1-[(4-methoxy)phenyl]methyl-8-quinolinol (III), protopine (IV) and 6-acetonyl-5,6-dihydrosanguinarine (V).

CONCLUSIONFive compounds were isolated from C. adunca for the first time.

Alkaloids ; chemistry ; isolation & purification ; Benzophenanthridines ; Berberine Alkaloids ; chemistry ; isolation & purification ; Corydalis ; chemistry ; Isoquinolines ; Molecular Structure ; Phenanthridines ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry

BACKGROUND: C1q nephropathy (C1qN) is a controversial diagnostic entity defined by Jennette and Hipp in 1985. The prevalence is very low and a few large scale studies have been reported. Application of the criteria for clinical diagnostics of C1qN may cause confusion with other glomerulonephropathies, such as minimal change disease (MCD) or focal segmental glomerulosclerosis (FSGS). In order to clarify the confusion with glomerulonephropathies, we did this study to identify the clinicopathological characteristics and the exact disease entity of C1qN. METHODS: A total of 5,258 kidney biopsies at Kangnam St Mary's Hospital were reviewed. Twenty three cases (0.44%) met the criteria of C1qN. Twenty eight cases showing dominant C1q deposits without electron dense depostis (EDD) grouped as C1q+EDD-, and previously diagnosed typical cases of MCD and FSGS were selected for this study. Four groups were compared to each other with regard to the clinical and pathological aspects of the disease. RESULTS: C1qN patients had an average age of 30.4 years. Eighteen were males and 5 were females. Eighty seven percent had proteinuria and 18% had hematuria. By electron microscopy analysis, 100% had mesangial EDD and 47.8% showed foot process effacement. C1qN had some significant differences compared with C1q+EDD-, MCD and FSGS. CONCLUSIONS: C1qN is clinically and morphologically different from MCD and FSGS. However, additional long term studies are needed to fully define C1qN from other glomerulonephritis with C1q deposits.
Biopsy ; Complement C1q ; Corneal Dystrophies, Hereditary ; Electrons ; Female ; Foot ; Glomerulonephritis ; Glomerulosclerosis, Focal Segmental ; Hematuria ; Humans ; Kidney ; Male ; Microscopy, Electron ; Nephrosis, Lipoid ; Phenanthridines ; Prevalence ; Proteinuria

A rapid, sensitive, and selective liquid chromatography-tandem mass spectrometry was developed for the simultaneous determination of lycorine and galanthamine, two major constituents in Lycoris radiata extract, in rat plasma. Liquid-liquid extraction with ethyl ether was carried out using diphenhydramine as the internal standard. The two bioactive alkaloids were separated on a Zorbax SB-C18 reserved-phase column (150 mm × 4.6 mm, i.d., 5 μm) by gradient elution using a mobile phase consisting of methanol with 0.1% formic acid (A) and water with 0.1% formic acid (B) at a flow rate of 0.6 mL/min. All analytes showed good linearity over a wide concentration range (r (2)>0.99) and the lower limit of quantification was 3.00 ng/mL for each analyte. The average extraction recovery of the analytes from rat plasma was more than 82.15%, and the intra-day and inter-day accuracy and precision of the assay were less than 12.6%. The validated method was successfully applied to monitoring the concentrations and pharmacokinetic studies of two Amaryllidaceous alkaloids in rat plasma after an oral administration of Lycoris radiata extract.
Amaryllidaceae Alkaloids ; pharmacokinetics ; Animals ; Chromatography, Liquid ; Galantamine ; pharmacokinetics ; Lycoris ; chemistry ; Male ; Parasympathomimetics ; pharmacokinetics ; Phenanthridines ; pharmacokinetics ; Plant Extracts ; chemistry ; pharmacokinetics ; pharmacology ; Rats ; Rats, Wistar ; Tandem Mass Spectrometry ; methods

In order to quantify the curing effects of phenanthridine on yeast prion, we introduced semi-denaturing agarose gel electrophoresis and fluorescence recovery after photobleaching techniques to quantify the curing effects of phenanthridine on yeast prion at the protein and cellular levels with the [PSI+] yeast strain expressing GFP-Sup35p (NGMC). The results showed that these two approaches could precisely quantify the curing effects of phenanthridine on [PSI+] cells. After a treatment for 1 through 5 days with phenanthridine, the curing rates of [PSI+] cells were 0%, 0%, 51.7%, 87.5% and 94.4%, respectively. Meanwhile, we quantified the sizes of Sup35p polymers in phenanthridine induced pink phenotype cells. The aggregation status in 1-2 days phenanthridine treated cells were similar to those in [PSI+] cells, while the aggregation status in 3-5 days phenanthridine treated cells were similar to those in [psi(-)] cells.
Computer Simulation ; Models, Biological ; Peptide Termination Factors ; metabolism ; Phenanthridines ; pharmacology ; Prions ; drug effects ; genetics ; metabolism ; Saccharomyces cerevisiae ; cytology ; drug effects ; metabolism ; Saccharomyces cerevisiae Proteins ; metabolism

To explore the effect of lycorine in inducing apoptosis of pulmonary carcinoma cell A549 and its mechanism. In the study, pulmonary carcinoma cell A549 were taken as the experimental subject and processed with different concentrations of lycorine (0, 0.5, 1.0, 2.0, 4.0 and 8.0 μmol x L(-1)). The MTT method was used to observe the cell proliferation. The apoptosis rate of A549 cells was determined by Annexin FITC/PI double staining. The microplate reader was used to detect the activities of Bcl-2, Bax and p53. The changes in mitochondrial membrane potential were measured by the flow cytometry. The expressions of apoptosis-related factors Bcl-2, Bax, p53 and Survivin were determined by Real-time PCR. The results showed that lycorine significantly inhibited the proliferation of A549 cells (P < 0.05), induced the apoptosis on A549 cells (P < 0.05), increased the activities of Bax and p53, reduced Bcl-2 activity and mitochondrial membrane potential, and notably changed the gene expressions of Bcl-2, Bax, p53 and Survivin (P < 0.05). In conclusion, lycorine can induce the apoptosis of A549 cells and be applied to treat pulmonary carcinoma. Its mechanism may be related to the activation of relevant factors in Bcl-2 signaling pathway.
Amaryllidaceae Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma ; drug therapy ; genetics ; metabolism ; physiopathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Phenanthridines ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; Signal Transduction ; drug effects ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the antitumor effect of lycorine on renal cell carcinoma ACHN cells and explore the possible mechanism.

METHODSWe used flow cytometry to examine the effect of lycorine on ACHN cell cycle and apoptosis. The cell proliferation, migration and invasion were assessed with MTS assay, wound healing assay, and Transwell assay, respectively. Colony forming assay was performed, and the mRNA and protein levels of Bax, Bcl-2, survivin, caspase-3, cyclin D1 and CDK4 were measured with qRT-PCR and Western blotting.

RESULTSLycorine obviously inhibited the proliferation of ACHN cells with an IC(50) of 24.34 µmol/L. Lycorine also induced apoptosis of ACHN cells, caused cell cycle arrest at G(0)/G(1) phase, and suppressed the colony forming ability of the cells in a dose-dependent manner. The migration and invasion of ACHN cells were significantly inhibited by 5 µmol/L lycorine. Lycorine up-regulated the mRNA levels of CDK4, Bax, caspase-3 while down-regulated the levels of survivin, Bcl-2 and Cyclin D1; the protein levels of CDK4 and Bax were increased and cyclin D1, Bcl-2 and surviving expressions were decreased, but caspase-3 expression showed no significant changes following the treatment.

CONCLUSIONLycorine has obvious antitumor effect against ACHN cells, suggesting its value as a new therapeutic agent for renal cell carcinoma.

Amaryllidaceae Alkaloids ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; Carcinoma, Renal Cell ; pathology ; Caspase 3 ; metabolism ; Cell Cycle Checkpoints ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase 4 ; metabolism ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Phenanthridines ; pharmacology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE: To find out the optimal conditions of human chromosomal analysis protocol in peripheral blood sample. METHODS: The experiments were made with the variations of phytohaemagglutinin, colcemid, ethidium bromide concentration and the variations of hypotonic solution exposure time. RESULTS: In the experiment on the optimal phytohaemagglutinin concentration, the highest mitotic index in the overall collected cells was obtained in phytohaemagglutinin concentration 15microL/ml. In the experiment on the concentration of mitotic arrestant colcemid, the proper chromosomal state that is meta phase stage and doesn't have many chromosomal crossings or tangles was obtained in colcemid concentration 0.05microg/ml. In the experiment on the optimal exposure time of hypotonic solution(0.075M KCl) treatment, the most suitable intervals between chromosomes were subtained in 20 minutes. In the experiment on the optimal concentration of ethidium bromide to obtain minute chromosomal bands, the best result was when ethidium bromide concentration 5microg/ml or 7.5microg/ml was addition to colcemid concentration 0.02microg/ml. CONCLUSION: The combination of phytohaemagglutinin 15microL/ml, colcemid 0.05microg/ml, hypotonic solution exposure time for 20 minutes is important to the collection of appropriate chromosome state in human chromosomal analysis using peripheral blood. In the case that needs to obtain minute bands, the elongated chromosomes are obtained when ethidium bromide 5microg/ml or 7.5microg/ml in addition to colcemid concentration 0.02microg/ml with the same conditions of phytohaemagglutinin and hypotonic solution.
Demecolcine ; Ethidium ; Humans ; Mitotic Index

The luminous intensity of dark variant (S1) separated from photobacterium phosphoreum (A2) was 1/10,000 less than that of wild-type. Ethidium bromide (EB) (0.6 mg/L), Mytomycin C (MC, 0.05 mg/L), 2-amino fluorene (2-AF, 1.0 mg/L) all could strongly induce reversion mutation for S1 within 24 h and increase reversion ratio significantly. The results of experiments indicated that these revertants had stable genetic characteristic and the mutation may take place at gene levels. The mutagenesis to S1 caused by EB, MC and 2-AF was detected and it may be used as a new rapid, simple and sensitive method for gene toxicant monitoring.
*Chemiluminescent Measurements ; Ethidium/pharmacology ; Ethidium/toxicity ; Luciferases/biosynthesis ; Mitomycins/pharmacology ; Mitomycins/toxicity ; Mutagens ; Mutation/*drug effects ; Photobacterium/*genetics ; Toxicology/methods ; Transcription, Genetic/drug effects ; Variation (Genetics)

Alleles and genotype frequencies and its distribution pattern for the highly polymorphic D1S80 locus were determined in a Korean population sample, especially in Kwangju and Chonnam, by using PCR followed by agarose gel electrophoresis with ethidium bromide staining, a procedure called the amplified-fragment-length polymorphism(Amp-FLP) technique. And the data were compared with the alleles and genotype frequencies of Finnish population, North American Caucasian, and Korean population(Seoul) which had been reported. In 203 unrelated Korean individuals 27 alleles and 84 genotypes were observed. The highest allele frequency was in allele M24(0.128) and tne next orders were inalleles M18(0.126), M29, M30, M31, and M28 and the other alleles showed relatively low frequencies. The highest frequency of genotype was in M18/M24 and the next order frequencies were M18/M30, M19/M27 M29/M29, and M18/M29. The homozyous genotypes were in 9 alleles such as M29, M24, M31, and M18, and most of heterozygous genotypes were composed of alleles of each homozygous genotypes and /or the other alleles, its composition of genotypes was 0.881(74/84), 183(0.901) of the 203 individuals alleles, its composition of genotypes was 0.881(74/84), 183(0.901) of the 203 individuals alleles, its composition of genotypes was 0.881(74/84), 183(0.901) of the 203 individuals were included. The VNTR D1S80 locus demonstrated a heterozygosity of 0.872. From the above results, VNTR D1S80 locus may be a powerful locus to identify individuals, however, the allele frequencies was not closely related to the genotype pattern, and the alleles of homozygous genotypes influenced on the chance of the recombination of the various genotypes. It is necessary to analyze the genotype distribution and the recombination pattern of alleles as well as alleles and genotype frequencies in each populations for statistical test at most highly polymorphic loci.
Alleles* ; Electrophoresis, Agar Gel ; Ethidium ; Gene Frequency ; Genotype* ; Gwangju ; Jeollanam-do ; Polymerase Chain Reaction ; Recombination, Genetic

Chromatography, High Pressure Liquid ; methods ; Drug Residues ; analysis ; Electrochemistry ; Ethidium ; analysis ; Luminescent Measurements ; methods ; Sensitivity and Specificity