OBJECTIVE: To establish a female rat CYP3A induction model which is used for studying CYP3A-mediated drug-drug interactions. METHODS: Female SD rats which were fed with standard diet were randomly divided into two groups, one was the control group, and the other one was the experimental group. The rats in the experimental group were administered respectively 20, 50, 80, 100, and 150 mg·g-1d-1 dexamethasone by gavage for 3 d to induce CYP3A enzymes. 24 h after 3 d, liver tissues were taken from both groups of rats and rat liver microsomes were prepared. CYP3A4 activity was determined with testosterone as the probe substrate. RESULTS: Testosterone metabolic rate was 31.68% in blank liver microsomes, and were 40.64%, 61.36%, 82.44%, 85.8%, and 83.36% in dexamethasone-induction group. Testosterone metabolic rate was improved by up to 160.23% after dexamethasone induction at 80 mg·g-1d-1 in female rats. CONCLUSION: Dexamethasone induction at 80 mg·g-1d-1 can significantly increase liver CYP3A enzyme activity in female SD rats, and the model can be used to study CYP3A-mediated drug-drug interactions.

This work was mainly studied the effects of the four alkaloids from Coptidis Rhizoma on the mouse peritoneal macrophages in vitro and preliminarily discussed the regulating mechanisms. The effect of alkaloids from Coptidis Rhizoma on the vitality of macrophages was measured by the MTT assay. The effect of alkaloids on the phagocytosis of macrophages was determined by neutral red trial and respiratory burst activity was tested by NBT. The expressions of respiratory-burst-associated genes influenced by alkaloids were detected by qRT-PCR. The conformation change of membrane protein in macrophages by the impact of alkaloids was studied by fluorospectro-photometer. Results showed that the four alkaloids from Coptidis Rhizoma could increase the phagocytosis of macrophages in different level and berberine had the best effect. Berberine, coptisine and palmatine had up-regulation effects on respiratory burst activity of mouse peritoneal macrophages stimulated by PMA and regulatory activity on the mRNA expression of PKC, p40phox or p47phox, whereas the epiberberine had no significant influence on respiratory burst. Moreover, alkaloids from Coptidis Rhizoma could change the conformation of membrane protein and the berberine showed the strongest activity. The results suggested that the four alkaloids from Coptidis Rhizoma might activate macrophages through changing the conformation of membrane protein of macrophages and then enhanced the phagocytosis and respiratory burst activity of macrophages. Furthermore, the regulatory mechanism of alkaloids on the respiratory burst activity of macrophages may be also related to the expression level of PKC, p40phox and p47phox.
Alkaloids ; pharmacology ; Animals ; Cells, Cultured ; Coptis ; chemistry ; Drugs, Chinese Herbal ; pharmacology ; Female ; Gene Expression ; drug effects ; Macrophages, Peritoneal ; drug effects ; Mice ; Phosphoproteins ; genetics ; metabolism ; Protein Kinase C ; genetics ; metabolism ; Rhizome ; chemistry

An RP-HPLC method for determination of luteolin from Elsholtzia blanda Benth. extracts in rats' plasma was established and the pharmacokinetics of luteolin in rats was studied. Drug blood samples from caudal vein were gotten after oral administration of luteolin. Plasma samples were determined by RP-HPLC after being deproteinized with trichloroacetic acid and extracted with ethyl acetate. The calibration curve was linear in the range of 0.37-47.27 microg x mL(-1). The limit of quantification was 0.37 microg x mL(-1). The method recovery of luteolin was 93%-99%. The extract recovery was 75%-85%. RSDs of intra-and inter-day precisions were less than 5%. The concentration-time curve of luteolin after oral administration of Elsholtzia blanda Benth. extracts was fitted to two compartment open model. Two factors analysis of variance were adopted in the evaluation of gender and time spots for collection of blood. The result suggested that the gender-based difference in blood-drug concentrations had statistical significance. The metabolite in blood was identified as galcuronide. The method is sensitive, specific, accurate, and is appropriate for determination of luteolin in vivo.
Administration, Oral ; Animals ; Area Under Curve ; Chromatography, High Pressure Liquid ; methods ; Female ; Lamiaceae ; chemistry ; Luteolin ; blood ; isolation & purification ; pharmacokinetics ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Sensitivity and Specificity ; Sex Factors

Objective To investigate the medical and pharmaceutical knowledge of patients with chronic diseases and analyse the influence factors of rational administration in patients ,to provid data to support the establishment of pharmaceutical service mode . Methods 386 cases of patients with chronic diseases were asked to finish the questionnaires for the medical and pharmaceutical knowledge ,and factors affecting the rational drug were analyzed by single factor and multiple factors Logistic regression analysis . Results Among the 386 patients ,cardiovascular and celebralvascular disease ratio was the highest(53 .3% ) ,followed by respiratory system diseases(13 .8% ) and the musculoskeletal system diseases (11 .50% );The averaged score of 386 patients was 1 .76 ± 0 .78 , medication knowledge was at a general level;single factor analysis results showed that there was significant difference(P<0 .05) between rational drug-use and abuse of drugs among patients in number ,form of payment ,marital status ,income ,education level , taking drug knowledge lectures ,combined treatment .Multivariate Logistic regression analysis showed that education level ,partici-pation in lectures ,drug combination ,disease species had a significant impact on the rational drug use among patients with chronic disease(P<0 .05) .Conclusion The pharmaceutical knowledge that patients with chronic disease mastered is unsatisfactory ;and unreasonable behavior of medication is common scence .Education level ,participation in lectures ,drug combination ,the number of diseases have great influence on the rational use of drugs in patients with chronic diseases .A kind of effective pharmaceutical service mode should be established for patients with chronic diseases by clinical pharmacists .This is a very meaningful work for rational ad-ministration .

OBJECTIVETo detect the sequence variations frequently found within the N- and C-terminal regions of Epstein-Barr virus (EBV) LMP1 gene in nasopharyngeal carcinoma (NPC) and to study the underlying mechanisms.

METHODSFresh tumor tissues were sampled from 63 patients with untreated NPC encountered in Affiliated Tumor Hospital of Sun Yat-sen University, Guangzhou. The N-terminal region of EBV LMP1 gene was amplified with nested polymerase chain reaction (PCR), followed by XhoI enzyme digestion. Nested PCR was also employed to detect the 30 base pairs deletion within the C-terminal region. Four-colored fluorescence terminator sequencing method was applied for bi-directional solid-phase sequencing of the 8 representative PCR products in 4 cases of NPC. The DNA sequence within the N- and C-terminal regions of LMP1 gene was then analyzed.

RESULTSThere were 4 patterns of sequence variations, namely, wt-XhoI/wt-LMP1 (4 cases, 6.3%), wt-XhoI and XhoI-loss/del-LMP1 (4 cases, 6.3%), wt-XhoI/del-LMP1 (5 cases, 7.9%) and XhoI-loss/del-LMP1 (50 cases, 79.5%), detected in the 63 studied cases. Sequence analysis showed that the EBV LMP1 gene had underwent non-synonymous and synonymous substitutions, as compared with the prototype of B95-8 cells. The ratio of non-synonymous to synonymous substitutions was 2.25.

CONCLUSIONSXhoI-loss/del-LMP1 is the predominant sequence variation pattern of EBV LMP1 gene in NPC from Guangzhou. The XhoI-loss variation seems to develop on top of del-LMP1. When compared with the EBV LMP1 gene in peripheral blood B-lymphocytes of virus carriers and in preinvasive epithelial lesions (reported previously), it is likely that the sequence variation patterns of LMP1 gene may represent 4 different phases of intrahost evolution of EBV during nasopharyngeal carcinogenesis.

Adult ; Aged ; Base Sequence ; DNA, Viral ; genetics ; Deoxyribonucleases, Type II Site-Specific ; genetics ; Female ; Gene Deletion ; Genetic Variation ; Herpesvirus 4, Human ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Nasopharyngeal Neoplasms ; virology ; Point Mutation ; Sequence Analysis, DNA ; Viral Matrix Proteins ; genetics

Cell lines of Bcap37 and Bcap37/MDR1 (the high P-glycoprotein (P-gp) expressing cell line) were used as model to investigate the different accumulations of (E)-2-(4-(diethylamino methyl) benzylidene)-5,6-dimethoxy-2,3-dihydroinden-one (BYZX) in the two kinds of cells. It was authenticated that whether BYZX was the substrate of P-gp. Meanwhile, the inhibitive effects of BYZX on the P-gp were investigated by determining the fluorescence intensity of rhodamine 123 in the model cells, with and without BYZX. A reversed-phase high-performance liquid chromatography (RP-HPLC) method was used to determine the accumulations of BYZX in the two cells. The results showed that the amount of BYZX accumulation in Bcap37/MDR1 cells were as many as those in Bcap37 cells (P > 0.05), and the concentrations of BYZX accumulated in the Bcap37/MDR1 cells did not increase when co-incubated with P-gp inhibitor verapamil. Furthermore, different concentrations of BYZX also had no effects on the efflux of rhodamine 123 (P > 0.05). These results indicated that there were no interactions between BYZX and P-gp. BYZX will not be pumped out of the cells, and it also not inhibited the P-gp. It was the useful advantage for its absorption.
ATP-Binding Cassette, Sub-Family B, Member 1 ; antagonists & inhibitors ; metabolism ; Cell Line, Tumor ; Drug Interactions ; Humans ; Indenes ; metabolism ; pharmacology ; Rhodamine 123 ; metabolism ; Verapamil ; pharmacology

OBJECTIVETo compare the tissue formation and the content of polysaccharide between the wild Dendrobium candidum and the cultured ones and to find any existed differences.

METHODBare-handed microtomy and photomicrography; The content of polysaccharide is determined by phenol-sulphuric acid method.

RESULT AND CONCLUSIONThere are no marked noticeable differences between the wild D. candidum and the cultured ones in terms of the tissure formation and the content of polysaccharide.

Dendrobium ; anatomy & histology ; chemistry ; growth & development ; Plant Stems ; anatomy & histology ; chemistry ; growth & development ; Plants, Medicinal ; anatomy & histology ; chemistry ; growth & development ; Polysaccharides ; analysis ; Tissue Culture Techniques

OBJECTIVETo explore if reduced number of circulating endothelial progenitor cells (EPCs) is a risk factor for patients with coronary slow flow (CSF).

METHODSThirty patients with CSF and 30 age and gender matched control subjects with normal coronary angiography were included in the study. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and plated on fibronectin-coated culture dishes. EPCs were characterized as adherent cells double positive for DiI-AcLDL-uptake and lectin-binding by converted fluorescence microscope (×200).

RESULTSSmoking, diabetes mellitus, hypertension and the levels of plasma lipoprotein profile were similar between the two groups (all P > 0.05). The number of EPCs was significantly lower in patients with CSF compared with control subjects (35.7 ± 5.9 vs.53.2 ± 5.9, P < 0.01). TIMI frame counts was correlated with circulating EPCs number (OR = 0.424, 95%CI 0.358 - 0.621, P < 0.01) and not associated with gender, age, smoking, diabetes mellitus, hypertension and the levels of plasma lipoprotein profile.

CONCLUSIONDecreased circulating EPCs is an independent risk factor for CSF.

Blood Circulation ; Blood Flow Velocity ; Case-Control Studies ; Cell Count ; Cells, Cultured ; Coronary Angiography ; Coronary Vessels ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Risk Factors ; Stem Cells ; cytology

OBJECTIVETo explore the influence of oxidized high-density lipoprotein (oxHDL) on the maturation and migration of bone marrow-derived dendritic cells (BMDCs) from C57BL/6J mice.

METHODSThe C57BL/6J mice bone marrow cell suspension was prepared and purified. Recombinant granulocyte-macrophage colony-stimulating factor (rmGM-CSF) and recombinant interleukin-4 (rmIL-4) were used to promote monocytes to differentiate and suppress lymphocytes. Then 50 microg/mL oxHDL was added to stimulate BMDCs, using 50 microg/mL high-density lipoprotein (HDL) as homologous protein control, PBS as negative control, and 1 microg/mL lipopolysaccharide (LPS) as positive control. The CD86 and MHCII expression rates were detected with fluorescence-activated cell sorting (FACS). Liquid scintillation counting (LSC) was used in mixed lymphocyte reactions (MLRs) to reflect the ability of BMDCs in stimulating the proliferation of homologous T cells. Levels of cytokines IL-12 and IL-10 were detected by ELISA. The cell migration was evaluated with the transwell system.

RESULTSCompared with PBS group, the expressions of CD86 and MHCII, counts per minute of MLRs, secretion of IL-12 and IL-10, and number of migrated cells in oxHDL group and LPS group significantly increased (all P<0.05), while the increment was less in oxHDL group than LPS group. The number of migrated cells in oxHDL group was about twice of that in HDL group.

CONCLUSIONOxHDL may promote the maturation and migration of BMDCs in vitro.

Animals ; Bone Marrow Cells ; cytology ; drug effects ; physiology ; Cell Differentiation ; drug effects ; Cell Movement ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; drug effects ; physiology ; Humans ; Lipoproteins, HDL ; metabolism ; pharmacology ; Lipoproteins, LDL ; metabolism ; pharmacology ; Mice ; Mice, Inbred C57BL

OBJECTIVETo observe the protective effects of total glycosides Rubus parviflolius (TGRP) on local cerebral ischemic.

METHODThe local cerebral ischemia in rat was made by middle cerebral artery occlusion(MACO). The infraction weight was determined by TTC stain. SOD, MDA, GSH and apoptotis were determined with different method respectively.

RESULTTGRP 20, 10 mg x kg(-1) ig markedly improved the abnormal nervous symptoms, incredsed the SOD, GSH activity and reduced contentes of MDA in brain of MACO rat, TGRP 20 mg x kg(-1) ig significantly decreased the numbers of apoptotic cells in ischemic cortex.

CONCLUSIONTGRP has protective effects against cerebral infraction, and its mechanism may be related to anti-apoptotis and free radical.

Animals ; Apoptosis ; drug effects ; Behavior, Animal ; drug effects ; Brain ; metabolism ; pathology ; Glycosides ; isolation & purification ; pharmacology ; Infarction, Middle Cerebral Artery ; metabolism ; pathology ; physiopathology ; Male ; Neuroprotective Agents ; pharmacology ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rosaceae ; chemistry