Integrative hepatoprotective efficacy comparison of raw and vinegar-baked Radix Bupleuri using nuclear magnetic resonance-based metabolomics.

Radix Bupleuri (RB), with a Chinese name Chaihu, is one of the most popular Traditional Chinese herbal drug. It can be baked with vinegar to afford vinegar-baked Radix Bupleuri (VBRB), which is used in Traditional Chinese Medicine (TCM) for liver diseases treatment. In the present study, nuclear magnetic resonance-based metabolomic approach was used to compare the liver protective effect of RB and two types of VBRBs, which were prepared by two kinds of vinegar. The contents of 14 metabolites in the liver of carbon tetrachloride (CCl4) treated mice were significantly altered in comparison with control group, and VBRB prepared by Shanxi vinegar showed best effect as revealed by the amount and regulatory degree of the perturbed metabolites. The metabolism pathways analysis showed that the liver protective effect was related with the energy metabolism, lipid metabolism, ketone body metabolism, glutathione metabolism, amino acids metabolism and nucleotide synthesis. The results presented here showed that metabolomic approach made it possible to disclose the subtle biological difference between two types of VBRB, which highlight the potential of metabolomic approach to quantitatively compare the pharmacological effect of the herbal drugs.

[Expression of peroxisome proliferator-activated receptor gamma in glioma].

OBJECTIVE: To investigate the expression and significance of peroxisome proliferators-activated receptor gamma (PPAR gamma) in human glioma. METHODS: Immunohistochemical staining for PPAR gamma was performed using biopsy specimens of human glioma of various histological types. Expression of PPAR gamma and GFAP in glioma cell lines SWO-38, U251 and SHG-44 were analyzed using Western blotting and reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: Immunohistochemical study showed that PPAR gamma was expressed in glioma tissues with positive rate of 37.5%. Western blotting and RT-PCR showed that PPAR gamma was expressed in both glioma cell lines SWO-38 and U251, but not in SHG-44 cells. However, high expression of GFAP was detected in SHG-44 cells. CONCLUSION: PPAR gamma is associated with carcinogens of glioma. Actived PPAR gamma by agonist may be a novel approach to the treatment of glioma.

[Expression of PTEN gene in human glioma cell lines U251 and SHG-44 and its effect on cell proliferation].

BACKGROUND & OBJECTIVE: Mutation or deletion of PTEN gene is related to a variety of tumors. PTEN gene abnormality is closely related to the tumorigenesis of glioma. This study aimed to investigate the expression of PTEN gene in human glioma cell lines U251 and SHG-44, and explore its effect on cell proliferation. METHODS: The expression of PTEN gene in U251 and SHG-44 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). The recombinant eukaryotic expression vector containing wild-type PTEN gene was transfected into U251 and SHG-44 cells by cation polymex. The stably transfected cells were selected by G418 and amplified. Cell morphology was observed under microscope. The effect of PTEN gene on cell proliferation was assessed. The expression of PTEN protein and glial fibrillary acidic protein (GFAP) were detected by Western blot and immunohistochemistry. RESULTS: Point mutation and deletion of PTEN mRNA were observed respectively in U251 and SHG-44 cells. The proliferation rates of U251 and SHG-44 cells were inhibited by 39.1% and 27.8% of control at 7 days after transfection. The expression of PTEN and GFAP were both increased. The stably transfected U251 cells differentiated toward astrocytes, but SHG-44 cells had no obvious morphologic changes. CONCLUSION: Restoring expression of wild-type PTEN could induce differentiation of glioma cells differently.

Expression of c-fos in rat brain as a prelude marker of central nervous system injury in response to methylmercury-stimulation.

OBJECTIVE: To probe into the prelude marker of central nervous system injury in response to methyl mercury chloride (MMC) stimulation and the signal transduction molecular mechanism of injury in rat brain induced by MMC. METHODS: The expression of c-fos mRNA in brain and the expression of c-FOS protein in cortex, hippocampus and ependyma were observed using reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods. The control group was injected with physiological saline of 0.9%, while the concentrations for the exposure groups were 0.05 and 0.5, 5 mg/kg MMC respectively, and the sampling times points were 20, 60, 240, 1440 min. RESULTS: The expression of c-FOS protein in cortex and hippocampus increased significantly, the accumulation of mercury in the brain induced by 0.05 mg/Kg MMC for 20 min had no significant difference compared with the control group. The mean value was 0.0044 mg/Kg, while the protein c-FOS expression had significant difference compared with the control group (P < 0.01). More sensitive expression occurred in hippocampus and cortex, but not in ependyma. Conclusion The expression of c-FOS protein in cortex and hippocampus can predict the neurotoxicity of MMC in the early time, and immediately early gene (IEG) c-fos participates in the process of brain injury induced by MMC.

Effects of mercury contaminated rice from typical chemical plant area in China on nitric oxide changes and c-fos expression of rats brain.

China is one of countries with the highest mercury production in the world. The Guizhou Province in Southwestern China is currently one of the world's most important mercury production areas. In order to study the neurotoxicity of rice from Qingzhen Chemical Plant area and probe into the signal transduction molecular mechanism of injury in rat brain stimulation by mercury contaminated rice. The rats were exposed to mercury contaminated rice for 20 d. Both of the measurements of NO and NOS were processed according to the protocol of the kit. The effect of Hg contaminated rice on the expression of c-fos mRNA in rat brain and the expression of c-FOS protein in cortex, hippocampus were observed using reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods. The results showed the neural transmitter NO and NOS in brain were significantly change between exposure groups and control group; the mercury polluted rice induced significantly the expression of c-fos mRNA; the c-FOS positive cells in hippocampus and cortex of exposure groups were significant different from control group (p < 0.01). It could be concluded that nitric oxide was involved in mercury contaminated rice induced immediate early gene c-fos expressions in the rat brain. Through food chain, local ecosystem and health of local people iave been deteriorated seriously by mercury. This serious situation will last a long period. In order to alleviate mercury pollution, more work needs to do.

Effect of methylmercury on some neurotransmitters and oxidative damage of rats.

In order to study the molecular mechanism of injury in rat organs induced by methylmercury, and the relationship between neurotransmitter and oxidative damage in the toxicity process of rat injury by methylmercury was studied. The control group was physiological saline of 0.9%, the concentration of exposure groups were 5 mg/( kg x d) and 10 mg/( kg x d) respectively. The content of AChE, ACh, NOS, NO, MDA, SOD, GSH-Px and GSH in different organs of rats were determined with conventional methods. The results showed that after exposure to methylmercury for 7 d, the mercury content in brain of exposure groups increased clearly and had significant difference compared with the control group (P < 0.01). In rat's brain, serum, liver and kidney, the content of ACh and AChE were all decreased; the content of NOS and NO were all increased; the content of MDA was increased compared with the control group, the exposure groups had significant difference (P < 0.01); the content of SOD, GSH and GSH-Px was decreased compared with the control group, the exposure groups had significant difference (P < 0.01). It could be concluded that methylmercury did effect the change of neurotransmitter and free radical. They participated in the toxicity process of injury by methylmercury. The damage of neurotransmitter maybe cause the chaos of free radical and the chaos of free radical may also do more damage to neurotransmitter vice versa.

[Antagonistic effects of selenium on the expression of c-fos in central nerval system of rat included by mercury contaminated rice].

The objective of this paper is to study the antagonisms between selenium and mercury and the effect of different species mercury on the brain injury. The expression of c-fos mRNA and c-FOS protein in rat brain induced by Hg-contaminated rice was observed by using reverse transcriptions polymerase chain reaction (RT-PCR) and immunocytochemical methods. The results show the Hg-contaminated rice induced significantly the expression of c-fos mRNA and c-FOS protein; selenium could antagonize mercury accumulative level in brain. Antagonistic effects of selenium on the expression of c-fos included by mercury and the molecule mechanism of the antagonisms between selenium and mercury was probed, too.

Rice from mercury contaminated areas in Guizhou Province induces c-jun expression in rat brain.

OBJECTIVE: Mercury (Hg), as one of the priority pollutants and also a hot topic of frontier environmental research in many countries, has been paid higher attention in the world since the middle of the last century. Guizhou Province (at N24 degrees 30'-29 degrees 13', E103 degrees 1'-109 degrees 30', 1 100 m above the sea level, with subtropical humid climate) in southwest China is an important mercury production center. It has been found that the mercury content in most media of aquatics, soil, atmosphere and in biomass of corns, plants and animals, is higher than the national standard. The present study aims to explore the influence of mercury pollution on the health of local citizens. METHODS: The effect of rice from two mercury polluted experimental plots of Guizhou Province on the expression of c-jun mRNA in rat brain and c-jun protein in cortex, hippocampus and ependyma was observed using reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemical methods. RESULTS: The results showed that the mercury polluted rice induced expression of c-jun mRNA and its protein significantly. Selenium can reduce Hg uptake, an antagonism between selenium and mercury on the expression of c-jun mRNA and c-jun protein. CONCLUSION: c-jun participates in the toxicity process of brain injury by mercury polluted rice, the expression of c-jun mRNA in brain, and c-jun protein in rat cortex and hippocampus can predict neurotoxicity of mercury polluted rice. People should be advised to be cautious in eating any kind of Hg-polluted foods. To reveal the relationship between c-jun induction and apoptosis, further examinations are required.

[Expression of c-FOS protein in rat hippocampus by inducing of mercury contaminated rice].

In order to probe into the biological effects of mercury in typical mercury contaminated area and try to apply the expression of immediately early gene c-fos in brain to early predict the neurotoxicity of mercury in typical polluted areas, the expression of c-FOS protein in rat hippocampus is observed using immunocytochemical methods. The results show the mercury pollutes rice induced significantly the expression of c-FOS protein in hippccampus;the antagonisis between selenium and mercury on the exposure process. It is suggested that c-fos can be used as an effective index of detecting and assessing neurotoxicology of mercury polluted areas.

[Effect of methylmercury on expression of immediate early gene c-jun mRNA in rat brain].

In order to probe into the early prediction molecular index and the signal transduction molecular mechanism of methyl mercury chloride (MMC) neurotoxicity, the expression of c-jun mRNA in rat brains induced by different concentration MMC for different times were observed by using reverse transcription polymerase chain reaction (RT-PCR) methods (the control group was physiological saline of 0.9%, the concentrations of expose groups were 0.05, 0.5, 5 mg x kg(-1) respectively, the sampling times were 20, 60, 240, 1440 min). The result showed the expression of c-jun mRNA in rat brains was prior to the accumulation of mercury, and the expression of c-jun mRNA in rat brains could early predict the neurotoxicity of MMC. IEG (c-jun) participated in the toxicity process of injury by MMC.

Analysis of alpha, beta, gamma-hexachlorocyclohexanes in water by novel activated carbon fiber-solid phase microextraction coupled with gas chromatography-mass spectrometry.

A fast and simple method for determination of alpha, beta, gamma-hexachlorocyclohexanes (HCHs) in water using activated carbon fiber-solid phase microextraction(ACF-SPME) were studied. Results showed the performance of adsorption and desorption of three HCHs on ACF were excellent. A wide linear range from 10 to 100 microg/L and detection limits of the ng/L level were obtained using ACF-SPME with GC-MS in selected ion monitoring(SIM) acquisition mode. The proposed method was also successfully applied for determination of three HCHs in tap water. Compared to commercial fibers, ACF showed some advantages such as better resistance to solvents, higher thermal stability, longer lifetime and lower cost. The data demonstrated that GC-MS with ACF-SPME is well suitable for the analysis of HCHs in water.

[Improvement of the determination method of benzene, toluene, ethylbenzene and xylene(BTEX) in water using activated carbon fiber solid-phase microextraction/gas chromatography-mass spectrometry(GC-MS)].

The methods of direct injection, carbon disulfide extraction and activated carbon fiber solid-phase microextraction/GC-MS, usually used in the determination of BTEX in water matrix, are compared and discussed. Experimental data of linearity, precision and limit of detection illustrate that the last one is better than the two other methods. This method was tested by the practical sample experiments and expected to be a simple and sensitive new method for the analysis of BTEX in water.