No abstract available.
Arthritis, Rheumatoid* ; Folic Acid* ; Metabolism* ; Methotrexate*

BACKGROUND: Methotrexate (MTX), one of the main drugs used to treat osteosarcoma, is a representative folic acid antagonist. Polymorphisms of various enzymes involved in the metabolism of MTX could contribute to differences in response to MTX in pediatric osteosarcoma patients. METHODS: Blood and tissue samples were obtained from 37 pediatric osteosarcoma patients who were treated with high-dose MTX therapy. The following 4 single nucleotide polymorphisms (SNPs) were analyzed: ATIC 347C>G, MTHFR 677C>T, MTHFR 1298A>C and SLC19A1 80G>A. Serial plasma MTX concentrations after high-dose MTX therapy and MTX-induced toxicities were evaluated. Correlations among polymorphisms, MTX concentrations and treatment-induced toxicities were assessed. RESULTS: Plasma MTX levels at 48 hours after high-dose MTX infusion were significantly associated with SLC19A1 80G>A (P=0.031). Higher plasma levels of MTX at 48 and 72 hours were significantly associated with MTX-induced mucositis (P=0.007 and P=0.046) and renal toxicity (P=0.002), respectively. SNP of SLC19A1 gene was associated with development of severe mucositis (P=0.026). CONCLUSION: This study suggests that plasma levels of MTX are associated with GI and renal toxicities after high-dose MTX therapy, and genetic polymorphisms that affect the metabolism of MTX may influence drug concentrations and development of significant side effects in pediatric patients treated with high-dose MTX.
Folic Acid* ; Humans ; Metabolism ; Methotrexate* ; Mucositis ; Osteosarcoma* ; Plasma ; Polymorphism, Genetic* ; Polymorphism, Single Nucleotide

Methotrexate (MTX) is a widely used immunosuppressive drug. Large-dose of MTX is used for the treatment of cancer while low-dose is used for the treatment of rheumatoid arthritis (RA). This study aimed to explore the effect of MTX on the urinary proteome of rats. MTX was given to rats orally to construct an MTX intragastric administration rat model. The urine of the rats were collected within 10 hours after giving MTX, and the urine proteins of the rats were analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). A total of 31 differential proteins were identified, of which 7 proteins were related to the effect MTX and the symptom of RA. The biological processes of some rats reflected the effect of MTX on the body's glutathione metabolism and the JAK/STAT signaling pathway, which indicated that urine proteins have the ability to reflect the effects of MTX on the body of rats. The spectrum of the differential proteins of each single rat showed that different individuals respond to the drug quite differently.
Rats ; Animals ; Methotrexate/metabolism* ; Proteome ; Chromatography, Liquid/methods* ; Tandem Mass Spectrometry/methods* ; Arthritis, Rheumatoid/drug therapy*

OBJECTIVETo explore the effect of Xinfeng Capsule (XC) on lipoprotein metabolism of rheumatoid arthritis (RA) patients.

METHODSTotally 180 RA patients were assigned to the experimental group and the control group by random digit table, 90 in each group. Patients in the experimental group took XC (three pills each time, three times daily), while those in the control group took Methotrexate Tablet (four tablets each time, once per week). One month consisted of one therapeutic course and all patients were treated for two therapeutic courses. A healthy control group consisting of 60 patients was also set up. Changes of lipoprotein indices, clinical efficacy, lipid metabolism, joint symptoms and signs, activity indicators were observed, and correlation analyses were performed.

RESULTSCompared with the healthy control group, expression levels of prealbumin (PA), globulin (GLO), high-density lipoprotein (HDL), apolipoprotein Al (Apo-A1) were lowered in RA patients (P <0. 05, P <0. 01). Correlation analyses showed that PA was negatively correlated with joint tenderness, morning stiffness time, disease activity score (DAS-28), C-reactive protein (CRP), interleukin (IL)-6, respectively. Total protein (TP) was negatively correlated with joint tenderness. GLO was negatively correlated with joint tenderness and DAS-28. HDL was negatively correlated with erythrocyte sedimentation rate (ESR) and endothelin (ET)-1. Apo-Al was negatively correlated with joint pain; Apo-B was negatively correlated with CRP; LDL was negatively correlated with morning stiffness time (P <0. 05, P <0. 01). Compared with before treatment, expression levels of PA, HDL, Apo-A1 , Apo-B, and serum IL-10 contents increased, and expression levels of ESR, CRP, IL-6, ET-1 , joint pain, joint swelling, morning stiffness time, and DAS-28 decreased in the experimental group (P <0. 05, P <0. 01). PA increased more after treatment than before treatment in the control group (P <0. 01). There was statistical difference in joint symptoms (except joint tenderness) and activity indices (except ET-1) in the control group (P <0. 05, P <0. 01). Compared with the control group after treatment, PA and HDL increased, ET-1 and duration of morning stiffness decreased in the experimental group (all P <0. 05).

CONCLUSIONSLipoprotein metabolic disorder exists in RA patients, and it is associated with disease activity. XC could obviously improve lipoprotein metabolism and joint symptoms.

Arthritis, Rheumatoid ; drug therapy ; metabolism ; Blood Sedimentation ; C-Reactive Protein ; Capsules ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Interleukin-10 ; Interleukin-6 ; Lipoproteins ; Lipoproteins, HDL ; metabolism ; Methotrexate

AIM: The most important side effect of methotrexate (MTX) is mucositis. The purpose of this study was to evaluate the effect of turmeric extract on intestinal damage and oxidative stress in rats receiving methotrexate. METHODS: Experiments were performed on male Wistar albino rats divided into six groups. First group received normal saline orally, the second group received turmeric extract (100 mg·kg(-1)) orally for 30 days, the third group received turmeric extract (200 mg·kg(-1)) orally for 30 days, the fourth group received a single dose of methotrexate (20 mg·kg(-1)) i.p. at day 30, the fifth group received turmeric extract (100 mg·kg(-1)) orally for 30 days and a single dose of methotrexate (20 mg·kg(-1)) i.p. at day 30, and the sixth group received turmeric extract (200 mg·kg(-1)) orally for 30 days and single dose of methotrexate (20 mg·kg(-1)) i.p. at day 30. Four days after methotrexate injection, animals were anesthetized, blood samples were taken to determine total antioxidant status (TAS) and jejunum samples were taken for glutathione peroxidase (GPx), superoxidase dismutase (SOD), catalase (CAT), aldehyde malondialdehyde (MDA), and histopathological assessment. RESULTS: Microscopic evaluation from intestinal tissues of the MTX treated group, showed severe villus shortening and blunting, inflammatory cell infiltration and hemorrhage in lamina propria, along with epithlial cell necrosis. Levels of SOD, GSH-Px and CAT decreased in the MTX received group, but increased significantly (P < 0.05) in the turmeric + MTX groups. MTX increased lipid peroxidation, however, turmeric decreased peroxidation significantly (P < 0.05). CONCLUSION These results suggest that turmeric extract may protect the small intestine of rats from methotrexate-induced damage. Turmeric effects could result from its antioxidant properties.
Animals ; Catalase ; metabolism ; Curcuma ; chemistry ; Glutathione Peroxidase ; metabolism ; Humans ; Intestinal Diseases ; chemically induced ; drug therapy ; enzymology ; metabolism ; Intestinal Mucosa ; metabolism ; Male ; Malondialdehyde ; metabolism ; Methotrexate ; adverse effects ; Oxidative Stress ; drug effects ; Plant Extracts ; administration & dosage ; Rats ; Rats, Wistar ; Superoxide Dismutase ; metabolism

This study is to observe the effects of methopterin on the activation and bone resorption function of murine osteoclasts, which were obtained by induction from bone marrow cell and purified to the purity of 70%-80%. The mechanism underlying the inhibitory effects of methopterin on inflammatory bone destruction was explored. MTT method was used to determine the effect of methopterin on the proliferation of osteoclasts. Flow cytometric analysis was used to determine the effect of methopterin on the apoptosis of osteocalsts. TRAP stain, bone resorption lacuna stain and measurement of lacuna area were executed to determine the effects of methopterin on the activation and function of osteoclasts. ELISA method was used to determine the effect of methopterin on the MMP-9 secretion from osteoclasts. RT-PCR method was used to determine the effect of methopterin on the mRNA expression of RANK and MMP-9 in osteoclasts. The results showed that methopterin (0.1-10 micromol x L(-1)) inhibited the proliferation of osteoclasts, methopterin (0.1-10 micromol x L(-1)) could inhibit the activation and bone resorption function of osteoclasts and induced the apoptosis of osteoclasts. Methopterin (0.01-10 micromol x L(-1)) also decreased the mRNA expression of RANK, but only at 1-10 micromol x L(-1) decreased the mRNA expression of MMP-9. These results indicated that there were intense relation between the inhibitory effects on the activation and function of osteoclasts and the inhibition of inflammatory bone destruction by methopterin.
Animals ; Antirheumatic Agents ; pharmacology ; Apoptosis ; drug effects ; Bone Resorption ; pathology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Male ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Methotrexate ; pharmacology ; Mice ; Mice, Inbred C57BL ; Osteoclasts ; cytology ; metabolism ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; metabolism

BACKGROUNDMagnetic targeting therapy may be a new method for the treatment of malignent tumors. The purpose of this study was to investigate the localization and distribution of ferrofluid microsphere of human serum albumin methotrexate (FM-HSA-MTX) carriers in the brain and to explore the magnetic targeting chemotherapy for malignant brain tumor.

METHODSNinety SD rats were divided into three groups: targeting group, non-magnetic targeting group, and control group. Synthesized FM-HSA-MTX carriers (MTX 25 mg/kg) were injected into the systemic circulation via the caudal vein (magnetic targeting group, n = 30). A 0.6 T magnetic field was placed around the right hemisphere. The non-magnetic targeting group (n = 30) was administered with FM-HSA-MTX without external magnetic field, meanwhile the control group (n = 30) was treated with MTX and a magnetic field. Random serial sacrifices (n = 10) were conducted at 15, 30 and 45 minutes after drug administration. Bilateral hemispheres were collected respectively, and analyzed for total MTX content.

RESULTSMTX content in the right hemisphere of the magnetic targeting group was significantly higher than that in the other two groups at 15, 30 and 45 minutes after drug administration (P < 0.05) No difference was seen between the non-targeting group and control group. In the magnetic targeting group, MTX returned to the peak level [(0.564 +/- 0.018) mg/g, q15-45 = 32.252, P < 0.05] 45 minutes after the injection but it deceased in the other two groups [non-magnetic targeting group: (0.060 +/- 0.015) mg/g, q15-45 = 9.245, P < 0.05, control group: (0.074 +/- 0.045) mg/g, q15-45 = 6.299, P < 0.05]. In the magnetic targeting group, the concentration of MTX in the right hemisphere was significantly higher than that in the left hemisphere (t45min = 21.135, P = 0.000) but no difference was observed between bilateral hemispheres in the other two groups (non-magnetic targeting group: t45min = 0.434, P = 0.670; control group: t45min = 0.533, P = 0.600).

CONCLUSIONIn the presence of the external magnetic field, FM-HSA-MTX can distribute successfully in the targeting areas of the brain.

Animals ; Antineoplastic Agents ; administration & dosage ; Brain ; metabolism ; Drug Carriers ; Magnetics ; Methotrexate ; administration & dosage ; pharmacokinetics ; Microspheres ; Rats ; Rats, Sprague-Dawley ; Serum Albumin ; administration & dosage ; pharmacokinetics

OBJECTIVE: This study examines the effects of tumor necrosis factor (TNF) blockade on markers of bone metabolism in peripheral blood from active rheumatoid arthritis (RA) patients. METHODS: Eighteen patients (16 women, 2 men) aged 50 years (range 37-63 years), with persistently active RA (mean disease duration 7 years) were studied. Most took methotrexate (mean dose 12.5 mg) and all except one received corticosteroid (mean dose 5.7 mg). Four were treated with etanercept, eight received adalimumab and six received infliximab. Before and six months after taking TNF blockers, blood was sampled to obtain peripheral blood mononuclear cells (PBMCs), and serum bone turnover markers and acute phase reactants were measured. PBMCs were seeded and cultured to produce osteoblastic lineage cells and osteoclasts. RESULTS: The formation of calcified nodules by osteoblastic lineage cells from PBMC increased from 205.7±196.3 µmol/well at the baseline to 752.5±671.9 µmol/well after TNF blockade (p<0.024). The serum levels of bone formation markers, including bone specific alkaline phosphatase and osteocalcin also increased. The number of circulating osteoclasts and area of bone resorption pits made by osteoclasts were reduced after TNF blockade. CONCLUSION: The activity of circulating osteoblastic lineage cells increased after TNF blockade, whereas peripheral osteoclastogenesis tended to be suppressed. This is the first study of cultured human peripheral osteoblastic lineage cells in RA patients. Given that peripheral bone formation is difficult to study using radiologic methods, culture of these cells may provide a new modality for studying bone metabolism in RA.
Acute-Phase Proteins ; Adalimumab ; Alkaline Phosphatase ; Arthritis, Rheumatoid ; Biological Therapy ; Bone Remodeling ; Bone Resorption ; Etanercept ; Female ; Humans ; Infliximab ; Metabolism ; Methotrexate ; Osteoblasts* ; Osteocalcin ; Osteoclasts* ; Osteogenesis ; Tumor Necrosis Factor-alpha*

Development of drug resistance in tumors during treatment is a major factor limiting the clinical use of anticancer agents. When tumor cells acquire resistance to anticancer drug, they show cross-resistance to other antitumor agents. In the present study, SNU-1 cell was induced adriamycin 10-7 drug resistance, SNU-1/ADR, in vitro culture system. We got the doubling time and number for viability test during 96 hours by MTT assay. To investigate the cross resistance of various anticancer drugs in human stomach cancer cell SNU-1 and SNU-1/ADR, We compared IC50 (drug concentration of 50% reduction) and the relative resistance (RR). SNU-1/ADR was expressed multidrug resistant with vinblastine (RR;>31.62), vincristine (RR;29.50), dactinomycin (RR;21.37), epirubicin (RR;17.78), daunorubicin (RR;14.12), adriamycin (RR;7.76), and etoposide (RR;4.46), and other drugs, 5-fluorouracil, cisplatin, cyclophosphamide, methotrexate, and calarubicin, have not cross resistant with adriamycin. There was double minute chromosome in SNU-1/ADR by karyotyping although this change was not seen in SUN-1.
Antineoplastic Agents ; Cisplatin ; Cyclophosphamide ; Dactinomycin ; Daunorubicin ; Doxorubicin ; Drug Resistance ; Epirubicin ; Etoposide ; Fluorouracil ; Humans ; In Vitro Techniques ; Inhibitory Concentration 50 ; Karyotyping ; Metabolism* ; Methotrexate ; Stomach Neoplasms* ; Stomach* ; Vinblastine ; Vincristine

Methotrexate(MTX) is a commonly used antimetabolite, which can be used in the treatment of a variety of diseases. However, hepatotoxicity in the use of MTX severely limits its clinical use. Therefore, how to prevent and treat hepatotoxicity of MTX has become an urgent clinical problem. This paper summarizes and analyzes relevant literatures on the prevention and treatment of hepa-totoxicity caused by MTX with traditional Chinese medicines and natural medicines in recent years. MTX-induced hepatotoxicity mechanisms include folate pathway, oxidative stress damage and adenosine pathway, of which oxidative stress theory is the main research direction. A total of 14 kinds of traditional Chinese medicine and natural medicine extracts including white peony root, and 21 kinds of natural monomer compounds, including berberine, play an anti-MTX-induced hepatotoxic effect by resisting oxidative stress, inhibiting inflammation and regulating signal pathways. According to current studies on the prevention and treatment of hepatotoxicity induced by MTX with traditional Chinese medicines and natural medicines, there are insufficiencies, such as partial and superficial mechanism studies, inadequate combination of experimental research and clinical practice, non-standard experimental design and lack of application of advanced technologies and methods. This paper systematically reviewed the effects and mechanisms of traditional Chinese medicines and natural medicines against hepatotoxicity induced by MTX and defined current studies and deficiencies, in the expectation of proposing new study strategies and directions and providing scientific basis for rational clinical use of MTX and development of new drugs against MTX hepatotoxicity.
Chemical and Drug Induced Liver Injury/prevention & control* ; Drug-Related Side Effects and Adverse Reactions ; Humans ; Liver/metabolism* ; Medicine, Chinese Traditional ; Methotrexate/toxicity* ; Oxidative Stress