1.Studies on the alkaloids from the herb of Corydalis adunca.
Yan-li TANG ; Ai-mei YANG ; Ying-shan ZHANG ; Han-qing WANG
China Journal of Chinese Materia Medica 2005;30(3):195-197
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
2.C1q Nephropathy: A Distinct Pathologic Entity.
Jung Ha SHIN ; Tae Eun KIM ; Kyo Young LEE ; Sang In SHIM ; Yeong Jin CHOI
Korean Journal of Pathology 2009;43(4):335-341
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
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Complement C1q
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Corneal Dystrophies, Hereditary
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Electrons
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Female
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Foot
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Glomerulonephritis
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Glomerulosclerosis, Focal Segmental
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Hematuria
;
Humans
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Kidney
;
Male
;
Microscopy, Electron
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Nephrosis, Lipoid
;
Phenanthridines
;
Prevalence
;
Proteinuria
3.Quantification of the curing effects of phenanthridine on yeast prion PSI+.
Zhengwei ZHONG ; Lijie WANG ; Hui XIE ; Hui LI ; Jianwei HE ; Youtao SONG
Chinese Journal of Biotechnology 2012;28(6):737-746
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
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Models, Biological
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Peptide Termination Factors
;
metabolism
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Phenanthridines
;
pharmacology
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Prions
;
drug effects
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genetics
;
metabolism
;
Saccharomyces cerevisiae
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cytology
;
drug effects
;
metabolism
;
Saccharomyces cerevisiae Proteins
;
metabolism
4.An LC-MS/MS method for the simultaneous determination of lycorine and galanthamine in rat plasma and its application to pharmacokinetic study of Lycoris radiata extract in rats.
Xin ZHOU ; Yue-bin LIU ; Shan HUANG ; Ying LIU
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(6):861-868
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
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pharmacokinetics
;
Animals
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Chromatography, Liquid
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Galantamine
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pharmacokinetics
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Lycoris
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chemistry
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Male
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Parasympathomimetics
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pharmacokinetics
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Phenanthridines
;
pharmacokinetics
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Plant Extracts
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chemistry
;
pharmacokinetics
;
pharmacology
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Rats
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Rats, Wistar
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Tandem Mass Spectrometry
;
methods
5.Antitumor activity of lycorine in renal cell carcinoma ACHN cell line and its mechanism.
Yi-Qiang HUANG ; Yi-Ming ZHANG ; Zhong JIN ; Xie-Zhao LI ; Chong-Shan WANG ; Kai XU ; Peng HUANG ; Chun-Xiao LIU
Journal of Southern Medical University 2016;36(6):857-862
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
6.Study on effect of lycorine in inducing apoptosis of pulmonary carcinoma cell A549.
China Journal of Chinese Materia Medica 2015;40(16):3278-3282
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
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pharmacology
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Antineoplastic Agents, Phytogenic
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pharmacology
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Apoptosis
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drug effects
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Carcinoma
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drug therapy
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genetics
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metabolism
;
physiopathology
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Cell Line, Tumor
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Cell Proliferation
;
drug effects
;
Drugs, Chinese Herbal
;
pharmacology
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Humans
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Lung Neoplasms
;
drug therapy
;
genetics
;
metabolism
;
physiopathology
;
Phenanthridines
;
pharmacology
;
Proto-Oncogene Proteins c-bcl-2
;
genetics
;
metabolism
;
Signal Transduction
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drug effects
;
Tumor Suppressor Protein p53
;
genetics
;
metabolism
;
bcl-2-Associated X Protein
;
genetics
;
metabolism
7.The Optimal Conditions of Chromosomal Analysis in Peripheral Blood.
Hee Gyung SON ; Young Il LEE ; Byung Joon JUNG ; Sang Hoon LEE ; Soon Ja KANG
Korean Journal of Obstetrics and Gynecology 2001;44(1):5-16
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
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Ethidium
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Humans
;
Mitotic Index
8.Reversion mutation in dark variants of luminous bacteria and its application in gene toxicant monitoring.
Journal of Huazhong University of Science and Technology (Medical Sciences) 2002;22(3):180-2
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
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Ethidium/pharmacology
;
Ethidium/toxicity
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Luciferases/biosynthesis
;
Mitomycins/pharmacology
;
Mitomycins/toxicity
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Mutagens
;
Mutation/*drug effects
;
Photobacterium/*genetics
;
Toxicology/methods
;
Transcription, Genetic/drug effects
;
Variation (Genetics)
9.Effect of Glutathione on Apoptosis Induced by Methyl Mercury Chloride.
Jung Ho YOUM ; Dai Ha KOH ; Keun Sang KWON ; Me Yae LEE
Korean Journal of Occupational and Environmental Medicine 2002;14(4):377-391
OBJECTIVES: This study was performed to evaluate the critical role of glutathione(GSH) in methyl mercury chloride(MeHgCl)induced cell apoptosis. METHODS: The effect of GSH in MeHgCl induced cell apoptosis was observed in mouse macrophage-derived RAW 264.7 cells in vitro. The cells were cultured in Dulbecco's modified Eagle's medium(DMEM). RESULTS: MeHgCl exerted a dose dependent cytotoxicity,as demonstrated by the MTT assay, which is an assay dependent partially on the mitochondrial function. Moreover, in the presence of NAC, a GSH precursor, the MeHgCl induced cytotoxicity was significantly decreased whereas BSO, a specific GSH synthesis inhibitor,increased the MeHgCl induced cytotoxicity.The MeHgCl induced DNA fragmentation and chromatin condensation was consistent with the morphological alterations. The MeHgCl treated cells exhibited increasing annexin V-FITC binding to the phos-phatidylserine(PS)translocated from the inner to the outer leaflet of the plasma membrane and those cells with NAC pretreatment significantly exhibited decreasing annexin V-FITC binding compared to the cells treated with MeHgCl only. However BSO pretreatment markedly exhibited the increasing annexin V-FITC binding. The MeHgCl treated cells generated ROS, which was evidenced by the oxidation of dihydroethidine and the generation of the fluorescent product, ethidium. In addition, BSO pretreatment further enhanced the extent of ROS generation caused by MeHgCl whereas NAC pretreatment decreased the amount of ROS generation. MeHgCl led to a dose dependent decrease in the GSH content. Although MeHgCl exposure significantly reduced the GSH level, those cells that had a NAC pretreatment contained a higher level of GSH compared to the cells treated with MeHgCl only. In contrast, BSO pretreatment futher enhanced the extent of GSH depletion caused by MeHgCl. CONCLUSIONS: These results indicate that MeHgCl reduced the GSH content and impaired the defense against oxidative damage caused by ROS formation in RAW 264.7 cells. It is possible that these factors leads to the activation of the apoptosis signaling pathway. Ultimately these results suggest that GSH plays a crucial role in protecting the activity against MeHgCl induced apoptosis.
Animals
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Antioxidants
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Apoptosis*
;
Cell Membrane
;
Chromatin
;
DNA Fragmentation
;
Ethidium
;
Glutathione*
;
Mice
10.The activity of antimelanocyte autoantibodies in vitiligo patients.
Yoon Kee PARK ; Seung Kyung HANN ; Min Seok SONG ; Jung Ku YOON ; Hyung Il KIM
Korean Journal of Dermatology 1991;29(3):391-398
Several observation suggest that the antimelanocyte autoantibodies could play a role in melanocyte destruction. Some experiments indicate that melanocyte antibodies from patients with vitiligo can kill melanocyte in vitro. In these experiments, we demonstrated that vitiligo patient's sera containing antimelanocyte antibodies can lyse cultured human melanocytes by complement activation. Melanocyte cytotoxicity was measured using the ethidium bromide/ acridine orange viability assay. Significant melanocyte cytotoxicity was seen in sera from patients with both active and inactive vitiligo(p<0.01). Melanocyte cytotoxicity measured with complement-mediated cytotoxicity decreased after systemic steroid treatment(p<0.05) ; however melanocyte cytotoxicity showed no significant change with systemic PUVA therapy.
Acridine Orange
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Antibodies
;
Autoantibodies*
;
Complement Activation
;
Ethidium
;
Humans
;
Melanocytes
;
PUVA Therapy
;
Vitiligo*