Juzen-taiho-to is a Kampo (Japanese and Chinese traditional) medicine, and is a nourishing agent, a so-called "Hozai" (in Japanese), that is used for improving disturbances and imbalances in the homeostatic condition of the body. This drug is administered to patients in various weakened conditions, including post-surgery patients and patients with chronic illnesses, where it can alleviate general symptoms such as extreme fatigue, pale complexion, loss of appetite, dry or scaly skin, night sweating, and dryness of the mouth. Currently, Juzen-taiho-to is often administered to cancer patients, and has been shown to possess various biological activities, such as enhancement of phagocytosis, cytokine induction, antibody production, induction of the mitogenic activity of spleen cells, anti-tumor effects when combined with surgical excision, anti-tumor effects with or without other drugs, and protection against the deleterious effects of anti-cancer drugs as well as radiation-induced immunosuppression and bone marrow toxicity.This article focuses on the antitumor and antimetastatic properties of Kampo formulations and describes the effect of Juzen-taiho-to and related formulations on tumor development, progression and metastasis in vivo.We also discuss the mechanism of the inhibitory action and the importance of the formulation and the constituent drugs in determining the efficacy.
4-Hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF) is representative of the Mailard reaction-derived reductones found in many foodstuffs. Influence of HEMF on iron ion-induced oxidative modification of human erythrocyte membranes and low density lipoprotein (LDL) under aerobic conditions was investigated.When human erythrocytes were incubated at 37°C for 24 h with HEMF alone, levels of thiobarbituric acid-reactive substances (TBARS) and non-ionic detergent C12E8-insoluble membrane protein aggregates in the membranes were unchanged. Levels of TBARS and protein aggregates in the membranes increased on treatment of the cells with Fe(III) ion at 37°C for 3 h were lowered in the presence of HEMF. Western blot analysis indicated that aggregates of band 3 protein induced by Fe (III) ion were decreased in the presence of HEMF, and the level of TBARS in LDL increased on treatment with Fe(III) of Fe(II) ion at 37°C for 24 h was also lowered in the presence of HEMF. The results indicate that HEMF protected the iron ion-induced oxidative modification of human erythrocyte membranes and LDL.
Tea and fruit juices are beverages consumed daily all over the world. The present study reports the inhibitory effects of these beverages on the activity of mammalian intestinal phenol sulfotransferases (P-STs).Green tea strongly inhibited the E. coli-expressed mouse intestinal P-ST activity in vitro. (-)-Epigallocatechin gallate (EGCG) was found to be the most potent inhibitor among the catechins tested (IC50=0.93 μM). (-)EGCG also inhibited the P-ST activity of the human colon carcinoma cell line, Caco-2. Kinetic analysis showed that the inhibition was competitive. Among fruit juices examined (apple, grape, grapefruit and orange), grape juice exhibited the most potent inhibitory action on the P-ST activity of mouse intestines and human colon carcinoma cells. The inhibitory activity of grape juice was located mainly in the skin and seels. Flavonols, such as quercetin and kaempferol, inhibited the P-ST activity at low concentrations. These observations suggest the possible inhibition of P-St activity in human intestines by green tea or grape juice.
Questions exist regarding tissue distribution of the β1 adrenergic receptor (β1-AR). The aim of this study was to investigate relative distribution patterns of the β1-AR at the protein level in a variety of human tissues by Western blot analysis. The specificity of anti-peptide antibodies was confirmed both by Western blot with recombinant β1-AR expressed as a membrane protein in E. coli and by immunoprecipitation of membranes from Sf9 cells infected with baculovirus to express the human recombinant β1-AR. β1-AR was found in all tissues examined. The relative amount of protein varied significantly between the tissues, from highest in lung and testis to very low in liver. β1-ARs were rather abundant in heart, kidney, placenta, spleen and thyroid. These results reveal unique distribution of β1-AR protein that suggests its tissue specific role. Moreover, our data demonstrate a high sensitivity of immunological detection that allows direct comparison of β1-AR subtype expression and could be used for receptor study in biopsies available in limited amounts, such as human heart biopsy.
β-Citrl-L-glutamate (β-CG) concentration was determined by HPLC during the differentiation of bovine lens epithelial cells into lens fiber cells in culture. β-CG increased from 1 to 4 weeks of culture and then decreased slightly, while α-crystallin, a marker of lens cell differentiation, increased rapidly 4 weeks after the culture and continued to increase gradually until week 11. In addition, the localization of β-CG was immunohistochemically examined using anti-β-CG antibody. Cells around lentoid bodies were stained with anti-β-CG antibody, whereas cells in the bodies were strained strongly with anti-γ-crystallin antibody. These findings suggest that β-CG accumulated immediately before the differentiation of the bovine lens epithelial cells into lens fiber cells and may play a role in regulating the differentiation of lens cells.
The present study investigated the effects of stable agonist for prostaglandin (PG) I2 receptor with PGI1 skeleton, SM-10906, on pro-inflammatory cytokine production by mouse peritoneal macrophages (PEMs) in comparison with PGE1 and PGI2. In mouse PEMs, SM-10906 and PGE1 slightly enhanced interleukin (IL)-6 secretion, but had no effects on tumor necrosis factor-α (TNF-α) or IL-1 production. SM-10906 concentration-dependently inhibited TNF-α, IL-1 and IL-6 releases from lipopolysaccharide-activated mouse PEMs, as with PGE1, PGI2 and cAMP analog. Additionally, SM-10906, PGE1 and PGI2 caused concentration-dependent accumulation of cAMP contents in mouse PEMs. It is concluded that PGI1 analog SM-10906 exerts anti-inflammatory effects on stimulated mouse PEMs by increasing in cAMP levels, as with E-series of PG.
The present study was undertaken to test if some cyclohexane dicarboximide derivatives may have a cardioprotective effect against hypoxia/reoxygenation injury. Isolated rat hearts were subjected to 20-min of hypoxiafollowed by 45-min reoxygenation, and their recovery of post-hypoxic cardiac contractile function was examined.Treatment with agents was carried out from 3 min after the onset of hypoxia to the end of hypoxia (17 min during hypoxia). Among the 17 compounds, 2-[4-[4-(4-chlorophenyl)-4-hydroxy-1-piperidinyl]butyl]hexahydro-1H-isoindol-1, 3(2H)-dione (ST-6) showed a significant enhancement of post-hypoxic contractile force. This was associated with attenuation of the releases of creatine kinase and purine nucleosides and based from the perfused heart. Hypoxia-induced increase in myocardial sodium and decrease in potassium ion content was suppressed by ST-6 treatment. The results suggest that ST-6 is capable of protecting the heart against hypoxia/reoxygenation injury possibly through a mechanism by which sodium overload during hypoxia is suppressed.
The active components of an aqueous extract of Sanguisorbae Radix, which possesses nitric oxide (NO) production-suppressing activity, were determined using macrophages that were activated by the addition of lipopolysaccharide. Significant inhibitory activity against the formation of both NO and inducible NO synthase, and NADPH-diaphorase activity, which is involved in NO generation, was shown by Sanguisorbae Radix fractions T-B and T-C. On further fractionation, the subfractions of T-B and T-C all showed high anti-NO activity.Sanguiin H-6, sanguiin H-11, 1, 2, 3, 4, 6-penta-O-galloyl-β-D-glucose, eugeniin and polymeric proanthocyanicdin were isolated from TB-3 and TC-4, and all were identified as exhibiting strong anti-NO activity. We have confirmed that sanguiin H-6 is the most active component of Sanguisobae Radix with respect to the suppression of NO production. It is suggested that tannin makes a prominent contribution to the biological activity of Sanguisorbae Radix.
The nematocidal activity of 38 quassinoids, C19 or C20 compounds isolated from Simaroubaceae, was measured using a species of Diplogastridae (Nematoda) to develop lead parasiticides. Of the various quassinoids tested, samaderines B and E displayed the most potent nematocidal activity with a minimum lethal concentration (MLC) of 2.0×10-5 M. The nematocidal activities of samaderines B and E were 15-fold greater than that of albendazole (3.0×10-4 M), 10-fold greater than that of thiabendazole (2.0×10-4 M) and 7.5-fold greater than that of avermectin (1.5×10-4 M). Thus, samaderines B and E may eventually be used as lead parasiticides. In light of the relationship between the structures of quassinoids and their nematocidal activities, those with potent nematocidal activity may require the elements mentioned. These results should help to further out understanding of nematocidal activity.
We analyzed the nucleotide sequences of the non-coding region of chloroplast DNA : the intergenic spacer between trnL (UAA) 3'exon and trnF (GAA). Two kinds of sequence, "type-1" and "type-2", were detected in 33 populations of Cannabis sativa. The length of the "type-1" fragment was 354 bp. In contrast, the "type-2" fragment from 3 populations was 353 bp long, with only one base deletion compared to "type-1." The fragment length from Humulus lupulus was 353 bp with a 1-bp deletion, and ten 1-bp substitutions compared to the sequences from C. sativa "type-1." Furthermore, we could clearly identify differences between C. sativa and H. lupulus using single-strand conformation polymorphism of PCR products (PCR-SSCP) analysis.
Hepatoprotective aliphatic glycosides 3-(S)-3-β-D-glucopyranosyloxybutanolide (1) and its congener, 3-(S)-3-β-D-glucopyranosyloxy-4-hydroxybutanoic acid (2) were isolated as major constituents from the whole plants of three Goodyera species, G. schlechtendaliana REICHB. fil., G. matsumurana SCHLTR. and G. discolor KER-GAWL. The structures of 1 and 2 have been determined by NMR, MS spectroscopic and chemical means. Compound 1 was converted into its methyl ester form (5) during the purification step, when the lactone ring was cleaved by catalysis of silica gel with the CHCl3-MeOH-H2O as a solvent. On the other hand, 1 was obtained in a high yield by the same purification procedure without MeOH. Based on this fact, a simple and economic method for the purification of 1 was confirmed. Compounds 1 and 2 were found to have a hepatoprotective effect on liver injury induced by carbon tetrachloride in primary cultured rat hepatocytes.
The neuropathological hallmarks of Alzheimer's disease (AD) are senile plaques, cerebrovascular β-amyloidosis, neurofibrillary tangles, and selective neuronal loss. β-Amyloid (Aβ) has been shown to cause vascular damage mediated by generation of reactive oxygen species and this damage is considered an early event in the development of AD. In this study, we determined the effect of pycnogenol, a potent antioxidant phytochemical, on Aβ-induced cellular injury. Pulmonary artery endothelial cells (PAEC) were exposed to Aβ for 24 h. Cell injury was assessed by measuring cell viability with methylthiazol tetrazolium (MTT) assay, and by determining the release of intracellular lactase dehydrogenase (LDH). Lipid peroxidation products of PAEC were determined by measuring thiobarbitric acid-reactive substances (TBARS). Exposure of PAEC to Aβ resulted in a decrease in cell viability, an increase of LDH release indicating membrane damage, and an elevated level of TBARS. Preincubation of PAEC with pycnogenol significantly minimized these changes. This study demonstrated that pycnogenol can protect vascular endothelial cells from Aβ-induced injury. The data suggest that pycnogenol may be useful for the prevension and/or treatment of vascular or neurodegenerative diseases associated with Aβ toxicity.
We developed a microelectrode technique to characterize the electrophysiological properties in Caco-2 cells, and used it to determine the mechanisms of absorption enhancers. The action of absorption enhancers on the apical membrane of Caco-2 cells was estimated by measuring the apical membrane potential (Vm) with the microelectrode. The Vm value of Caco-2 cells in Hanks' balanced salt solution containing 0.5 mM K+ was 18.9±0.8 mV (n=217), and the apical membrane resistance was 49.4±1.1 MΩ (n=160). In the electrophysiological study with absorption enhancers, laurylmaltoside (LM) markedly decreased the Vm value, while sodium glycocholate(NaGC) moderately reduced this value, and EDTA did not affect the value. These findings might be associated with their action sites, plasma membrane or tight junction in Caco-2 cell monolayers. In influx and transport studies with these absorption enhancers, LM enhanced the influx of furosemide, which is transported via both the transcellular and paracellular routes into Caco-2 cells, and enhanced its transport to the basolateral side of Caco-2 monolayers more than that of 5(6)-carboxyfluorescein (CF), a paracellular marker. EDTA did not increase the influx of furosemide, and enhanced the transport of furosemide and CF across Caco-2 cell monolayers to the same extent. In contrast, NaGC only slightly increased the influx of furosemide and did not enhance the transport of either furosemide or CF across the Caco-2 monolayers in this study. These findings were well correlated with the effects of these absorption enhancers on the electrophysiological parameters. Therefore, the microelectrode technique might be useful for evaluating the action of absorption enhancers in the plasma membrane at an intact cell level.
The uptake of fractionated heparin was examined in the absence and presence of anionic proteins such as acetylated low density lipoprotein (Ac-LDL) and maleylated bovine serum albumin (Mal-BSA) to characterize the scavenger receptors involved in the uptake of fractionated heparin in isolated rat Kupffer cells. The uptake of fractionated heparin was completely inhibited by Ac-LDL and dextran sulfate, but only partially by Mal-BSA.Kinetic analysis revealed that the binding capacity (Bmax) of the Mal-BSA-insensitive receptor was significantly larger than that of the Mal-BSA-sensitive one, though their dissociation constants (Kd) were not significantly different. The apparent internalization rate constant (kint, app) was significantly larger for the Mal-BSA-sensitive receptor than for the Mal-BSA-insensitive one. Thus, the scavenger receptors involved in the uptake of fractionated heparin in Kupffer cells can be classified into two types, in terms of sensitivity to Mal-BSA. Mal-BSA-sensitive receptors have been characterized in macrophages and classified as class A. The Mal-BSA-sensitive one found in Kupffer cells in this study may belong to class A, while the Mal-BSA-insensitive one has been little characterized elsewhere.
Falconensones A and B are a new type of yellow pigment with structural similarity to retinoic acid isolated from the mycelial extract of ascomycetous fungi, Emericella falconensis or Emericella fruticulosa. In the present study we show that falconensone A alone induced apoptosis of HL60 human leukemia cells, while falconensone B, the 4'-nor-methyl derivative of falconensone A, had much lower activity. The synthetic derivatives of falconensone A, falconensone A p-bromophenylhydrazone and falconensone A dioxime, were more potent than natural falconensone A and B as far as the induction of apoptosis was concerned. The induction of apoptosis by the falconensones correlated with their inhibition of cell growth. In addition, falconensones A and B, and falconensone A dioxime, increased the generation of intracellular reactive oxygen species, while falconensone A p-bromophenylhydrazone was inactive. These results suggest that falconensone A, falconensone A p-bromophenylhydrazone and falconensone A dioxime are potential new apoptosis-inducing agent. The enhanced generation of reactive oxygen species in cells may be involved in apoptosis induced by falconensone A and falconensone A dioxime, but not by falconensone A p-bromophenylhydrazone. It is also suggested that the methyl residue at the 4' position of the falconensone A cyclopentenone ring may be essential for the induction of apoptosis. Based on these results, falconensone A and its derivatives may be clinically useful in the treatment of some leukemias.
Follistatin (FS, an activin-binding protein) and activin A (homodimer of inhibin βA chain) promote and inhibit cell proliferation in rat liver, respectively. The roles of activin AB (heterodimer of inhibin βA and βB) and activin B (homodimer of inhibin βB) in rat liver have not been elucidated yet. In this study, we examined, by reverse-transcriptase polymerase chain reaction (RT-PCR) analysis, whether the levels of FS, inhibition βA and βB mRNAs change in the carbon tetrachloride induced rat liver regeneration model. The analysis was made in an hour-by-hour manner during the early stage of liver injury. There are 2 types of FS mRNA, FS-288 and FS-315, and the levels of both had begun to increase at 3 h, were maximal at 6 h, remained constant up to 12 h, and thereafter gradually decreased. The inhibin βA mRNA had started to decline at 3 h, reached its lowest level at 6 h, partly returned at 12 h, and remained constant up to 48 h. The inhibin βB mRNA level had begun to increase at 1 h, was maximal at 3 h, remained constant up to 24 h, and returned to the original level at 48 h. These results indicate that FS and activin A may act reciprocally in liver regeneration, and also sggest that activin AB and B may play roles in liver regeneration that differ from that of activin A.
The umbrella pine Sciadopitys verticillata seeds were found to contain a substantial amount (16.7 nmol/g) of sciadonic acid (all-cis-5, 11, 14-eicosatrienoic acid)-containing 2-monoacylglycerol, i.e., 2-sciadonoylglycerol (2-eicosa-5', 11', 14'-trienoylglycerol). Because the structure of 2-sciadonoylglycerol closely resembles that of 2-arachidonoylglycerol, the endogenous natural ligand for the cannabinoid receptor, we examined whether or not 2-sciadonoylglycerol exhibits cannabimimetic activity using NG108-15 neuroblastoma×glioma hybrid cells which express the cannabinoid CB1 receptor. We found that 2-sciadonoylglycerole induces rapid transient elevation of intracellular free Ca2+ concentration in NG108-15 cells through a cannabinoid CB1 receptor-dependent mechanism similar to the case of 2-arachidonoylglycerol, yet the activity of 2-sciadonoylglycerol was apparently lower than that of 2-arachidonoylglycerol. The activity of 2-sciadonoylglycerol was detectable from 3-10 nM, reaching a maximum at around 10 μM. To our knowledge, this is the first report showing the occurrence of a cannabimimetic monoacylglycerol in higher plants.
We examined the effects of 9 kinds of Kampo medicines, which are clinically used for the treatment of hypertension, on anesthetized rats with increases in arterial blood pressure, heart rate and peripheral blood flow induced by theophylline (5 mg/kg, i.v.) that were partially or completely mediated by endogenous catecholamines. Each Kampo medicine (1 g/kg) was intraduodenaly administered. Shinbu-to caused a severe disturbance of the arterial blood pressure. Saiko-ka-ryukotsu-borei-to, Oren-gedoku-to, San'o-shanshin-to and Dai-jyoki-to had hypotensive effects, while Hachimi-jio-gan, Gosha-jinki-gan, Dai-saiko-to and Choto-san did not have such an effect. Moreover, Saiko-ka-ryukotsu-borei-to attenuated the heart rate. In Oren-gedoku-to, San'o-shashin-to and Dai-jyoki-to, a reduction in peripheral blood flow was observed. These results suggest that Saiko-ka-ryukotsu-borei-to, Oren-gedoku-to, San'o-shashin-to and Dai-jyoki-to are ameliorative to the hypertension in sympathetic system dominance and Shinbu-to is occasionally dangerous to it.
We studied the antioxidant activities of calcium antagonists against autoxidation in rat brain homogenates.The homogenates were incubated for 30 min at 37°C with or without a calcium antagonist and subsequently assayed for lipid peroxide content. Percent inhibition of the lipid peroxidation was used as an index of the antioxidant effect. Dihydropyridine calcium antagonists exhibited concentration-dependent (3-300 μmol/l) inhibitory effects against lipid peroxidation. The relative order of antioxidant potency and associated IC50 values (μmol/l) of the calcium antagonists for inhibition of the lipid peroxidation were as follows : nifedipine (51.5)>barnidipine(58.6)>benidipine (71.2)>nicardipine (129.3)>amlodipine (135.5)>nilvadipine (167.3)>nitrendipine (252.1)>>diltiazem (>300)=verapamil (>300). These results suggest that some dihydropyridine calcium antagonists show antioxidant properties. The antioxidant effects of the calcium antagonists may contribute to their pharmacological actions.
MET-88, 3-(2, 2, 2-trimethylhydrazinium) propionate, suppresses carnitine synthesis by inhibiting (γ-butyrobetaine hydroxylase. The purpose of this study was to clarify the effects of suppression of carnitine synthesis on carnitine and lipid contents in tissues. MET-88 (50, 100, 200 or 400 mg/kg/d) was administered orally to male SD rats for 10, 30 or 60 d. Total carnitine and lipid (triglycerides, non-esterfied fatty acids) contents were measured in heart and liver. In both tissues, treatment with MET-88 dose-dependently decreased total carnitine levels, and the reduction reached the plateau state after 30 d at each dose. MET-88 had no effect on lipid content in the heart, but increased the lipid content in the liver at the highest doses. Treatment with MET-88 at 400 mg/kg for 60 d resulted in no pathologic findings in the histological study, and also had no effect on parameters of liver function such as glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase as judged from the results of blood biochemical analysis.We concluded that long-term treatment with MET-88 decreased the carnitine content to a constant level in both heart and liver, but had no effect on lipid contents in the heart, although it affected lipid metabolism in the liver.
This paper investigates the post-surgical pharmacokinetics of methotrexate (MTX) in the plasma, the cerebrospinal fluids (CSF), and the spaces created by tumor removal (STR) in patients with glioblastoma, during hyperosmotic disruption of the blood brain barrier (HODBBB) and intra-arterial chemotherapy with MTX. Eight Japanese patients with glioblastoma, three with open STRs and five with closed STRs, received a total of thirteen courses of HODBBB and intra-arterial combination chemotherapy with MTX. The patients were initially administered mannitol, then the anticancer drugs were infused into the carotid artery. Samples of blood and CSF from the STRs were obtained. MTX concentrations were measured by fluoroscence polarization immunoassay and the pharmacokinetic parameters of MTX in plasma and CSF were estimated. The plasma concentrations of MTX peaked at the end of drug infusion, and then decayed bi-exponentially during the remainder of the treatment period. The CSF concentration of MTX in the STR peaked 2 h after drug administration, then mono-exponentially decreased. The area under the concentration-time curve (AUC) for plasma and CSF MTX concentrations increased in parallel with the MTX dose. In patients with open STRs, the mean AUC of MTX in CSF was 4.44% of that found in plasma, while in patients with closed STRs, the mean was 61.2% of that found in plasma.In the latter group, the MTX administered using HODBBB and intra-arterial chemotherapy was maintained in the STRs for long periods.
We have studied the effects of monosialoganglioside (GM1)-containing cationic liposomes with a cationic cholesterol on the liposome-mediated gene transfection into mammalian culture cells. The results showed that both cationic liposomes with either a cationic cholesterol derivative of a hydrophobic amino head group (I) and a hydrophilic amino head group (II) promoted the transfection of luciferase plasmids (pGL3) into HeLa and CHO-K1 cells more than the control cationic liposomes without GM1. In addition, we found that cationic liposomes with a cationic cholesterol derivative (II) were about ten times as effective as that by commercially available cationic liposome Lipofectin. Confocal fluorescence microscopy showed that the liposome/DNA comples was transferred more efficiently into the target cells by the GM1-containing liposomes thant by the liposomes without GM1. In proportion to the above results, free antisense DNAs were also more efficiently transferred into the nucleus of the target cells by the GM1-containing liposomes. When there was 100 mM galactose in the transfection medium, the luciferase activity by the GM1-containing liposomes was reduced to the level of the control liposomes. The results suggest that GM1-containing cationic liposomes with a cationic cholesterol derivative of a hydrophobic amino head group or a hydrophilic amino head group should significantly increase the transfection efficiency of plasmid DNAs and antisense DNAs by galactose receptor-mediated endocytosis. This means that the GM1-containing liposomes described here should be very promising for gene transfection in vitro.
There are no reports on microbial contamination of repeatedly used lubricant (84-87% glycerin containing 0.02% benzalkonium chloride) for non-touch urethral catheters in intermittent self-catheterization. In this work, we evaluated microbial contamination of in-use lubricant and its prevention. Between September and December, 1996, microbial contamination and water activity of in-use lubricants in sheathed and lubricated non-touch chatheters connected to a tube used by 46 outpatients at our hospital was examined. Microbial contamination was detected at 5 to 2.6×108 colony-forming units (CFU)/ml in 14 (30.4%) of 46 samples. With higher water activity of the lubricant (a higher dilution rate of the lubricant with water), a higher concentration of microbial contamination was observed. In 3 (21.4%) of the 14 patients using contaminated lubricant, urine samples showed the same microbial species as those detected in the lubricant. To prevent microbial contamination of the lubricant, dilution of the lubricant with water, as a result of repeated use, should be avoided.
This report investigates the pharmacokinetics of cytosine arabinoside (Ara-C), methotrexate (MTX), nimustine (ACNU) and valproic acid (VPA) in cerebrospinal fluid (CSF) during CSF perfusion chemotherapy. A 28-year-old Japanese woman with disseminated glioblastoma was, on admission, on a stable oral regimen of prolonged-release VPA tablets (DepakeneTM-R), 400 mg twice a day, for seizure control. Twelve courses of CSF perfusion chemotherapy with Ara-C, MTX, and ACNU were administered. Plasma samples and CSF samples via Ommaya reservoirs were obtained during the eleventh course of treatment. The Ara-C and ACNU concentrations were measured by HPLC. The MTX and VPA concentrations were measured by fluorescence polarization immunoassay.During CSF perfusion chemotherapy, the highest CSF concentrations of Ara-C, MTX, and ACNU were observed at the end of the perfusion and decreased in a monoexponential pattern. The half-lives of Ara-C, MTX, and ACNU were 2.65, 3.52, and 0.71 h, respectively. No anticancer drugs were detectable in plasma during CSF perfusion chemotherapy. Before CSF perfusion chemotherapy, the free VPA concentration in plasma was 14.4% of the total VPA concentration. The mean total and free VPA concentrations in plasma were 78.0±0.8 and 10.9±0.3 μg/ml, respectively. The free VPA concentrations in plasma and in CSF were of similar values. At the end of perfusion, the lowest free VPA concentration in CSF was 30.3% of that at the initiation of perfusion. The free VPA concentrations in CSF at 3, 7, 23, and 47 h after the end of perfusion were 79.8, 94.5, 100.9, and 100.9% respectively of that at the initiation of perfusion. During CSF perfusion chemotherapy, the ratio of free VPA concentrations to the total VPA in CSF was 86.3±6.9%. The VPA concentrations in CSF rapidly decreased during the CSF perfusion but recovered to pre-treatment levels within 7h.