Effects of 27 natural products on drug metabolism genes in channel catfish (Ictalurus punctatus) cell line.

Pregnane X receptor (PXR) as a ligand dependent transcription factor, is capable of regulating gene expression of cytochromes P450 and transporters involved in xenobiotic/drug metabolism and elimination. Due to the species differences in the regulatory specificity of PXR, gene regulation should not be extrapolated from mammal to fish without research data.The aim of present study was to investigate the effect of 27 natural products on PXR, CYP3A30 and MDR1 genes in channel catfish (Ietalurus punetaus) kidney cells (CC-K). The results showed that bisdemethoxycurcumin, glycyrrhetnic acid, rotenone, artemisinin, dihydroartemisinin, ligustilide and matrine strongly induced the mRNA levels of PXR. Additionally, the up-regulation of CYP3A30 gene ran parallel with PXR gene after the treatment of demethoxycurcumin, glycyrrhetnic acid, artemisinin, matrine, baicalein, schisantherin A, ligustilide, and dihydroartemisinin. Moreover, we found that natural products schisandrin A, schisandrin B, schisandrol A, and schisandrol B significantly up-regulated the mRNA level of MDR1 gene.Our work with a view to provide experimental data support for further research, which will make for the rational application of natural products in channel catfish, such as to avoid adverse herb-drug interactions or accelerating the residue elimination of chemical medicine.

Wuzhi capsule and haemoglobin influence tacrolimus elimination in paediatric kidney transplantation patients in a population pharmacokinetics analysis: A retrospective study.

WHAT IS KNOWN AND OBJECTIVES: Tacrolimus is widely used for kidney transplantation in children. However, the narrow therapeutic window and considerable interindividual and intraindividual variabilities make tacrolimus untoward to design an optimum dosage for paediatric personalized medicine. Our research aims to establish the tacrolimus population pharmacokinetics (PPK) of Chinese paediatric kidney transplantation patients and to distinguish covariates impacting variabilities. METHODS: Chinese paediatric kidney transplantation patients treated with tacrolimus between January 2014 and April 2018 from Children's Hospital of Fudan University were retrospectively analysed. A total of 51 Chinese paediatric kidney transplantation patients were analysed using non-linear mixed effects modelling (NONMEM). The effects of population characteristics, biological features and drug combination were assessed. The final PPK model was evaluated using visual inspection of routine diagnostic plots and the internal validation method of bootstrap. RESULTS: Our data met the condition of a one-compartment model, and the final model was CL/F = 32.7 × (WT/70)0.75  × (1 - WZ × 0.341) × (HGB/97)-0.508 ; V/F = 1890 × (WT/70) × (POD/57)0.816 , where WT, WZ, HGB and POD were weight, Wuzhi capsule (extracted from schisandra sphenanthera, whose primary efficient constituents are schisantherin A, schisandrol B, schisandrin etc, and often used to treat drug-induced hepatitis in Chinese organ transplant patients), haemoglobin and post-transplant day, respectively. WHAT IS NEW AND CONCLUSION: The tacrolimus PPK model in Chinese paediatric kidney transplantation patients was developed, and Wuzhi capsule and haemoglobin influence tacrolimus elimination in paediatric kidney transplantation patients.

[Effect of forchlorfenuron on fruit morphology and lignans content of Schisandra chinensis].

The effect of plant growth regulator forchlorfenuron (CPPU) 1 x 10(-6), 0.67 x 10(-6), 0.5 x 10(-6) on fruit morphology and effective components lignans was studied. Those morphologies were the combination of four basic morphological changes. The result showed, diametre were increased and longitudinal diametre of fruits were inhibited by foliage fertilizers including CPPU. At the same time, 1 000-grain weight and yield showed the varying degrees increase under CPPU. The order of the degree was 0.5 x 10(-6) > 1 x 10(-6) > 0.67 x 10(-6). Six lignans content of Schisandra chinensis of different harvest time and different CPPU processing groups were determined, the results showed that lignans accumulation occurred mainly in periods of premature the half mature fruiting stages. Under the 0.67 x 10(-6) CPPU treatment, schisandrol B, schisandrin B, schisandrin C content of S. chinensis showed different increase.

Two polycyclic geranylhydroquinone-derived metabolites from roots of Arnebia hispidissima (Lehm.) DC.

A phytochemical investigation of the least polar organic extracts of Arnebia hispidissima (Lehm.) DC. roots has led to the isolation of two unique polycyclic geranylhydroquinone-derived metabolites, arnebacene (1) and arnebidin (2), along with some known phenolic metabolites viz., arnebin-7 (3) and vanillic acid (4). The chemical identification of the new isolated compounds, including their relative stereochemistry, was achieved via spectroscopic analyses, including 2D NMR, and spectral comparison with related compounds. A biosynthetic pathway is proposed for the new compounds on the basis of their structure-relationship with previously isolated metabolites.

Screening of herbal components for attenuating amiodarone-induced hepatotoxicity on gel-entrapped rat hepatocytes.

Amiodarone (AMD) is a hepatotoxic drug that has been widely used as a class III antiarrhythmic drug. Because, to date, only a few kinds of protectants are able to reduce AMD hepatotoxicity, this article utilized gel-entrapped rat hepatocytes to screen effective protectants from a series of herbal compounds for their effects against AMD-induced toxicity. Herbal compounds, including matrine, silibinin, glycyrrhizic acid, schisandrin B, epigallocatechin gallate and anisodamine, were cotreated with AMD to assess their protective effect, whereas vitamin E, which has been shown to be protective in rats, was selected as a control. It was found that vitamin E, as with its function in rats, provided the best protection in gel-entrapped rat hepatocytes, whereas silibinin, a major component of silymarin, could largely reduce AMD-induced hepatotoxicity, performing a similar function as silymarin in rats. The results illustrated that gel-entrapped hepatocytes may reflect the protective effects of drugs and serve as a reliable model for screening hepatoprotectants. Moreover, matrine, a widely used monomer of the traditional Chinese medicine, Sophora flavescens, for treatment of arrhythmia, was evidenced to show some effective protections against AMD hepatotoxicity. Taken together, gel-entrapped rat hepatocytes may provide a platform for screening effective candidates from the herbal component library.

Effect of acupuncture on target tissue distribution of Schisandra lignans.

BACKGROUND: Recently, the combination of acupuncture and Chinese medicine as a practical strategy to treat diseases is receiving considerable attention worldwide as they are usually found to exhibit intriguing therapeutic effectiveness. The current study aimed to study the adjunct effect of acupuncture on target tissue distribution of schisandra lignans when acupuncture is combined with Schisandra chinensis. METHODS: A simple and reliable high performance liquid chromatography-electrospray tandem-mass spectrometry (HPLC-ESI-MS) method for simultaneous analysis of three bioactive lignans (schisandrin, deoxyschisandrin and schisandrin B) in rat tissues was established. Using this analytical method we evaluated whether acupuncture had a synergistic effect on the tissue distribution of schisandra lignans. RESULTS: Tissue concentrations of the three lignans in the group receiving acupuncture were significantly higher than those in the schisandra only group, suggesting that acupuncture may potently increase tissue concentrations of schisandra lignans. The highest concentrations of the three lignans occurred in the liver compared with other tissues, and tissue concentrations in the heart, spleen, lungs and kidneys were increased by 315%, 203%, 250% and 224%, respectively. In addition, retention times of the lignans in tissues were prolonged for a relative long time. CONCLUSIONS: Our date indicate that the combined use of acupuncture and Schisandra chinensis could produce a synergistic effect which could play a beneficial role on promoting the tissue distribution of lignans. This has supported our initial hypothesis. The HPLC-MS method showed good sensitivity in quantifying the three schisandra lignans in different tissues.

Comparison of intracellular zinc signals in nonadherent lymphocytes from young-adult and elderly donors: role of zinc transporters (Zip family) and proinflammatory cytokines.

Intracellular zinc homeostasis is crucial in regulating the inflammatory/immune response at any age. It is tightly regulated by zinc transporters that control influx, efflux and compartmentalization of zinc within the cells. Specific methods for detecting the age-related differences in intracellular zinc signaling are poorly described. We report a novel assay induced after the in vitro zinc addition in peripheral blood mononuclear cells (PBMCs) and in lymphocytes from young and old donors in the absence/presence of in vitro zinc depletion (using EDTA). The intracellular labile zinc variations are monitored over time by flow cytometry using Fluozin-3 AM probe. The best curve fit of the data is calculated using a nonlinear regression model defined as follows: pr3/[1+Exp(-pr1-pr2*Xt)]. Pr1 depends on the initial free zinc value (time 0); pr2 describes the rate of the speed in reaching the maximum intracellular free zinc concentration; pr3 represents the maximum intracellular zinc increment (plateau curve); Xt is the time course. Age-related intracellular free zinc variations occur in PBMCs and lymphocytes incubated in EDTA-supplemented medium. The higher plateau of the curve (pr3) was observed in younger subjects. An up-regulation of Zip genes (Zip1, Zip2, Zip3), influencing zinc influx, is more pronounced in the young than old donors. Interleukin-6 and tumor necrosis factor-α overproduction was enhanced in old individuals, suggesting the presence of more marked zinc deficiency and chronic inflammation. In conclusion, the determination of intracellular zinc signals induced by in vitro zinc addition using logistic parameters may be useful to estimate the rate of intracellular zinc homeostasis and its role in inflammatory/immune response in aging.

Aerobic remediation of petroleum sludge through soil supplementation: microbial community analysis.

The effect of soil concentration on the aerobic degradation of real-field petroleum sludge was studied in slurry phase reactor. Total petroleum hydrocarbons (TPH) and polycyclic aromatic hydrocarbons (PAHs) showed effective removal but found to depend on the soil concentration. Aromatic fraction (48.12%) documented effective degradation compared to aliphatics (47.31%), NSO (28.69%) and asphaltenes (26.66%). PAHs profile showed efficient degradation of twelve individual aromatic compounds where lower ring compounds showed relatively higher degradation efficiency compared to the higher ring compounds. The redox behaviour and dehydrogenase activity showed a linear increment with the degradation pattern. Microbial community composition and changes during bioremediation were studied using denaturing gradient gel electrophoresis (DGGE). Among the 12 organisms identified, Proteobacteria was found to be dominant representing 50% of the total population (25% of γ-proteobacteria; 16.6% of ß-proteobacteria; 8.3% of α-proteobacteria), while 33.3% were of uncultured bacteria and 16.6% were of firmicutes.

Schisandrin B protects against solar irradiation-induced oxidative stress in rat skin tissue.

Schisandrin B (Sch B) and schisandrin C (Sch C), but not schisandrin A and dimethyl diphenyl bicarboxylate, protected rat skin tissue against solar irradiation-induced oxidative injury, as evidenced by a reversal of solar irradiation-induced changes in cellular reduced glutathione and α-tocopherol levels, as well as antioxidant enzyme activities and malondialdehyde production. The cytochrome P-450-mediated metabolism of Sch B or Sch C caused ROS production in rat skin microsomes. Taken together, Sch B or Sch C, by virtue of its pro-oxidant action and the subsequent eliciting of a glutathione antioxidant response, may prevent photo-aging of skin.

Biomonitoring potentials of polycyclic aromatic hydrocarbons (PAHs) by higher plants from an oil exploration site, Nigeria.

Higher plants sampled from a moderately polluted exploration site were analyzed with gas chromatography-mass spectrometry to determine the occurrence and sources of polycyclic aromatic hydrocarbons (PAHs). The concentrations of the sum of 28 target PAHs (designated as Σ(28)PAH) in the leaves of higher plant samples ranged from 365 to 2870 µg/kg with an average of 1430 µg/kg. The majority of the target compounds were detected except 9,10-diphenyl anthracene and dibenzo(a,h)anthracene, which were below the detection limits in most plant samples. In addition, the concentrations of the 2- and 3-ring PAHs, especially naphthalene and its alkylated derivatives, were generally higher than the 4-, 5- and 6-ring PAHs. Based on the results obtained in the present study, higher plants, especially non-woody annual and perennial plants could serve as good phytoremediators for a PAHs polluted sites.

[Research on liposoluble ingredients of Quchiling].

OBJECTIVE: To study the liposoluble ingredients of Quchiling (LQ), which enter the blood and the brain,and to confirm the active ingredients of LQ in vivo. METHOD: Serum pharmacochemistry and gas chromatography mass spectroscopy were used to analyze ingredients of LQ entering the blood and the brain. RESULT: There were eleven ingredients of LQ to enter the blood and six ingredients of LQ to enter the brain. CONCLUSION: It is confirmed that eleven ingredients of LQ entered the blood, which are beta-asarone, schisandrol A, schisandrol B, deoxyschisandrin, schisandrin B, schisantherrin A, schisantherrin B, schisantherrin C, delta-cadinene, delta-cadinol and calamendiol in the blood, and that six ingredients are beta-asarone, schisandrol A, schisandrol B, deoxyschisandrin, schisandrin B and calamendiol in the brain.

Effect of salinity on biodegradation of polycyclic aromatic hydrocarbons (PAHs) of heavy crude oil in soil.

The spillage of crude oil in the soil damages the environment. Polycyclic aromatic hydrocarbons (PAHs) are one of the crude oil components that may be harmful for living organisms. PAHs can disappear from the environment by volatilization and biodegradation. The effect of different NaCl concentrations (0%-5%) on PAHs reduction from the heavy crude oil-contaminated soil was studied. Our results showed that increasing NaCl concentration in soil had decreasing effect on total crude oil and PAHs reduction. The biodegradation of total crude oil was higher in 0% NaCl (41%) while higher total PAHs reduction was observed in 1% NaCl (35%). The lower total crude oil and PAHs reduction were observed in 5% NaCl (12% and 8% respectively). Phenanthrene, anthracene and pyrene reduction were higher in 1% NaCl, while fluoranthene and chrysene reduction were higher in 0% NaCl.

[Enzyme kinetics of schizandrin metabolism and sex differences in rat liver microsomes].

To study the enzyme kinetics of schizandrin metabolism in different gender in rat liver microsomes, liver microsomes were prepared from male or female rats. Schizandrin was incubated with rat liver microsomes. Schizandrin and its metabolites were isolated and identified by HPLC-UV method. Vmax, Km and Cl(int) of schizandrin in male and female rat liver microsomes were (21.88 +/- 2.30) and (0.61 +/- 0.07) micromol x L(-1) x min(-1) x mg(-1) (protein), (389.00 +/- 46.26) and (72.64 +/- 13.61) micromol x L(-1), (0.0563 +/- 0.0007) and (0.0084 +/- 0.0008) min x mg(-1) (protein), respectively. The major metabolites of schizandrin in female and male rat liver microsomes were 7,8-dihydroxy-schizandrin (M1) and 7, 8-dihydroxy-2-demethyl schizandrin (M2b), respectively. Ketoconazole, quinidine, and orphenadrine had different level effects on schizandrin metabolism in both male and female rat liver microsomes, and cimetidine still had some inhibitory effect in male liver microsomes. CYP3A and CYP2C11 may be the main P450 enzymes in schizandrin metabolism and their difference in rat liver microsomes may be the main reason for the sex difference of metabolic enzyme kinetics and metabolites of schizandrin in rats.

DNA damage in eelpout (Zoarces viviparus) from Göteborg harbour.

The relationship between DNA damage and the exposure of marine organisms to environmental contaminants was examined in the Göteborg harbour area. This research is part of a wider ecotoxicological study planned to evaluate the biological impact of chemical contamination in the River Göta estuary, following a bunker oil (10-100 tonnes) spill occurred in June 2003. Here we present data on the DNA strand breaks derived using the comet assay and the presence of apoptotic cells using the diffusion assay in nucleated erythrocytes of the eelpout (Zoarces viviparus) from the study area and at a clean reference site. Polycyclic aromatic hydrocarbon metabolites were also analyzed in the bile of exposed fish. The results showed a high level of damaged DNA, paralleled by a peak in bile PAH metabolites, in fish from the most impacted site, 3 weeks after the oil spill. A significant recovery was observed in specimens from the spill site, 5 months later, but not in fish caught in the middle part of Göteborg harbour, which is chronically subjected to heavy chemical pollution. The levels of apoptic cells did not show any marked variations, but a significant recovery was observed in fish from the oil impacted site 5 months after the spill.

[Distribution and physiology of microorganisms in petrochemical oily sludge of plant]. / Osobennosti raspredeleniia i fiziologicheskogo sostoianiia mikroorganizmov nefteshlama--otkhoda neftekhimicheskogo proizvodstva.

The occurrence, vertical distribution, and the physiological state of microorganisms in a petrochemical oily sludge deposit were studied. The total number and the number of viable microbial cells at depths of 0.2 and 3 m were about 10 and 10(8) cells/g dry wt. sludge. Most microbial cells taken from the middle (1 m deep) and the bottom (3 m deep) sludge horizons showed a delayed colony-forming ability, which suggested that the cells occurred in a hypometabolic state. The relative number of microaerobic denitrifying microorganisms steeply increased with depth. The amount of microorganisms tolerant to 3, 5, and 10% NaCl and capable of growing at 7 and 40 degrees C varied from 10(2) to 10(8) CFU/g dry wt. sludge. Petrochemical oily sludge was found to maintain the growth of heterotrophs, among which the degraders of oily sludge and ten different individual polycyclic aromatic hydrocarbons were detected. The occurrence of highly adaptable microorganisms with an adequate metabolic potential in the petrochemical oily sludge deposit implies that its bioremediation is possible without introducing special microorganisms.

Bacillus naphthovorans sp. nov. from oil-contaminated tropical marine sediments and its role in naphthalene biodegradation.

A Bacillus sp., designated as strain MN-003, was isolated as the dominant cultivatable naphthalene-degrading organism from oil-contaminated tropical marine sediments. Strain MN-003 is strictly aerobic, rod-shaped, Gram-positive, catalase positive, oxidase negative, and forms endospores. Strain MN-003 grew at salinities ranging from 0.28 to 7.00% and temperatures ranging from 15 to 41 degrees C. Phylogenetic analyses reveal that strain MN-003 is most similar to Bacillus sp. VAN14, with a 16S rRNA sequence identity of 97.9%. Based on taxonomic and 16S rRNA data, strain MN-003 was named Bacillus naphthovorans sp. nov. When grown with naphthalene as sole carbon source, strain MN-003 had a maximal specific growth rate (mu(max)) of 0.32 +/- 0.03 h(-1), and a half-saturation constant (K(S)) of 22.3 +/-4.2 microM. A batch study of the tropical marine sediments enriched with naphthalene showed that cells of the Bacillus genus grew to become dominant members of the microbial community. The bacilli comprised 39.5 +/- 6.5% of the microbial fraction after 20 days of enrichment.

Localization of deposited polycyclic aromatic hydrocarbons in leaves of Plantago.

After deposition to foliage, polycyclic aromatic hydrocarbons (PAHs) may remain on the leaf surface, accumulate in the cuticular wax, or diffuse into the remaining interior of the plant. In a field study, the location of deposited PAHs in the leaves of two Plantago species was determined. To this aim, leaves of Plantago major and Plantago media were divided into three fractions. First, the leaves were washed (wash-off fraction), then cuticular wax was extracted (wax fraction). Finally, the remaining leaf material was extracted (interior fraction). The presence of PAHs could be demonstrated in all three fractions. For both plants, the distribution of PAHs over the three fractions changed with molecular weight (mol wt) of the PAHs. The wash-off fraction increased with increasing molecular weight, likely because high molecular-weight PAHs occur predominantly bound to particles, which can be readily washed off from the leaves. In contrast, the amount of PAHs detected in the interior of the leaves decreased with increasing molecular weight. This can be explained by a slow desorption of the PAHs from the particles and a low diffusion rate of the larger molecules. This study shows that washing reduces the amount of high molecular-weight PAHs on plant surfaces. Therefore, washing of leafy vegetables is important to minimize human dietary intake of PAHs.

[Protective effect of schisanhenol (SAL) on oxygen free radical induced injury of spleen lymphocytes in mice].

In the present paper the effects of Sal, an active component isolated from Kadsura Longipenduculata Finet et Gagnep, on oxygen free radical induced injury of murine spleen lymphocytes in vitro and in vivo were investigated. The results showed that Sal at 1 x 10(-4) mol.L-1 significantly inhibited the decrease of GSH content induced by Fe(2+)-Vit C and the increase of MDA formation induced by Fe(2+)-Cys in murine spleen lymphocytes. The decrease of membrane fluidity of lymphocytes under lipid peroxidation induced by Fe(2+)-Vit C was prevented by Sal. The damage of the lymphocyte surfaces induced by Fe(2+)-Vit C was also markedly reduced by Sal. Administration of Sal 20 mg.kg-1.d-1 for 8 d significantly increased the GSH content of the spleen lymphocytes in mice under high oxygen-stress. Meanwhile, the compensative increase of SOD activity in the spleen lymphocytes was reversed. The results suggest that Sal may protect murine spleen lymphocytes from the injury induced by oxygen free radicals.

Degradation of crude oil by Acinetobacter calcoaceticus and Alcaligenes odorans.

Two types of Indian crude oil (Bombay High and Gujarat) were tested for their biodegradability by Acinetobacter calcoaceticus and Alcaligenes odorans. Acinetobacter calcoaceticus S30 and Alc. odorans P20 degraded Bombay High crude oil by 50% and 45%, while only 29% and 37% of Gujarat crude oil (heavy crude oil) was degraded by these isolates, respectively. Acinetobacter calcoaceticus and Alc. odorans in combination degraded 58% and 40% of Bombay High and Gujarat crude oils, respectively, which were significantly higher than that of by individual cultures. Acinetobacter calcoaceticus S30 degraded more of the alkanes fraction than the aromatics fraction of both crude oils. GC fingerprinting of alkane fraction showed major degradation of heptadecane (C17), octadecane (C18), nonadecane (C19), eicosane (C20), docosane (C22), tricosane (C23) and tetracosane (C24) of crude oil, while the Alc. odorans P20 degraded alkanes and aromatics equally. The asphaltenic component increased in both types of crude oil after biodegradation . The two strains grew very well on n-alkane up to C33 as well as on pristane (branched-chain alkane) but could not grow on cycloalkanes. Acinetobacter calcoaceticus S30 could not grow on pure polycyclic aromatic hydrocarbon (PAH) compounds except naphthalene but Alc. odorans P20 could grow on anthracene, phenanthrene, dibenzothiophene, fluorene, fluoranthene, pyrene and chrysene.

Isolation and characterization of four polycyclic aromatic hydrocarbon degrading bacteria from soil near an oil refinery.

Four bacterial strains (I-IV) capable of optimum growth on 0.1% naphthalene, anthracene or a mixture of naphthalene and phenanthrene were isolated from soil near an oil refinery. Two isolates (I and II) were identified as belonging to the genus Micrococcus, while strains III and IV were identified as Pseudomonas and Alcaligenes respectively. All the isolates were found to bear high molecular weight plasmid DNA (isolate I and IV 89%, II 67.5% and III 92.1% of lambda DNA), which is presumed to aid in the metabolism of polycyclic aromatic hydrocarbons. The strains also showed appreciable growth at high concentrations of NaCl (up to 7.5%).