As a member of growth arrest and DNA damage inducible gene family,GADD45αparticipats in the regulation of cell cycle,cell senescence,cell survival and apoptosis. GADD45αplays a critical role in the responses to cell injury induced by a variety of factors including cell stress and genotoxic chemicals. Different transcription factors and proteins are involved in transcriptional regulation of GADD45αgene. GADD45αprotein has been implicated in the regulation of genomic stability related cellular responses through interaction with other proteins. Genotoxicity test systems based on the char?acteristics of GADD45α in regulation of cell function,can be applied to the detection of potentially genotoxic compounds,which provides new ideas and methods about genotoxicity assessment. The molecular mechanism and research progress of GADD45α in genotoxicity test are summarized in this article.

In recent years,researches on drugs concentrated in the ecosystem have become a hot topic. Environmental pharmacology is a new subject studying the source and fate of drugs in the environment. It is found that a lot of pharmaceuticals and pharmaceutically active compounds are released into the environment by the pharmaceutical industry,hospitals and agricultured activities. Drugs in the environment not only affect the ecosystem,which leads to bacterial resistance,but also affect the balance of flora in the biofilm. Furthermore,such drugs may induce epigenetic changes, which may leave to the body vulnerable to diseases. The enrichment of endocrine disruptors can inter?fere with the growth and function of the human reproductive system. China is becoming concerned about environmental drugs. The development of environmental pharmacology can help clarify the interac?tions between drugs and the environment while contributing to public health and food safety in China.

Microbiome is a novel research field related to human health,agriculture,bio-energy and the environment. Gut microbiome has received much attention from researchers recently. Studies have shown that gut flora is related to some diseases,such as digestive disease(inflammatory bowel disease),metabolic disease(type 2 diabetes), cardie-cerebral vascular disease(Parkinson disease). Traditional Chinese materia medica(TCMM) has long been used as a tonic and taken in a large amount. Gut flora has an effect on pharmacology and toxicology of TCMM after entering the gastroin?testinal tract. This article is intended to review recent researches on microbiome,common detection techniques and the relationship with hepatotoxicity induced by Polygonum multiflorum Thunb.,scutel?laria baicalensis that directly affects the intestinal tract,nephrotoxicity induced by Rhizoma Alismatis and pneumonia induced by Xiao-Chaihu-Tang.

There is emerging evidence from clinical studies that the existence of liver cancer stem cells(CSCs)or tumor initiating cells is responsible for the high recurrence rates of tumor,generation of metastasis,and resistance to therapeutic regimens after therapy. Here,the characteristics of liver CSCs,clinical manifestation,molecular signaling Wnt/β-catenin,signal transducers and activators of transcription 3,NANOG,annexin A3/c-Jun N-terminal kinase,and chapter four-transmembrane 4 L six family member 5/CD44 in liver CSC self-renewal were briefly reviewed. In addition,potential targets for drug therapy were analyzed,providing some reference for drug discovery that selectively target liver CSC self-renewal signals.

OBJECTIVE To investigate the forms of carbamazepine and its metabolites in the gastric juice,blood and urine samples from a poisoned patient and its metabolic pathways. METHODS The gastric juice,blood and urine samples were obtained from a patient who was rushed to the Hospital of Prevention and Treatment for Occupational Diseases to be treated for an overdose of carbamazepine for about six hours. The samples were analyzed by gas chromatography-mass spectrometry. The metabolites were identified from their mass spectra by comparison with spectra in NIST 98 libraries. The metabolic pathways were inferred by fragmentation regularities of mass spectra and the published data. RESULTS The results showed that only M1 was found in the gastric juice sample. Four metabo?lites,such as iminostilbene(M1),9-methyl-acridine(M2),9-propyl-acridan(M4)and M3,were found in the blood sample and thirteen metabolites,including M1,M2,M4,acridine(M5),acridone(M6), 9,10-dihydro Acridine (M7), 1a,10b-dihydro-6H-dibenz[b,f]oxireno[d]azepine-6-carboxamide (M8),9-acridinemethanol (M9),10,11-dihydrodiol-carbamazepine (M10),2-methyl-acridone (M12),4-methyl-acridone(M13),undetermined M3 and M11,were found in the urine from the poi?soned patient. The forms of carbamazepine metabolites in the gastric juice ,blood and urine were different. Little carbamazepine was found in the urine after metabolism. CONCLUSION Based on the analysis by gas chromatography-mass spectrometry and identification of NIST 98 libraries,the metabolites could be identified accurately,which can help analyze the metabolic mechanism and pathways of carbam?azepine.

OBJECTIVE To investigate the change of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),intercellular adhesion molecules(ICAM-1),matrix metalloproteinases 2 (MMP-2) and MMP-9 contents in cultured pulmonary microvascular endothelial cells (PMVECs) in rats after perfluoroisobutylene (PFIB) exposure. METHODS PMVECs were separated and purified using a modified method of implantation of pulmonary tissues. After identification,PMVECs were divided into the normal control group and the PFIB-exposed groups(n=3). The PFIB-exposed groups inhaled PFIB at the concentration of 200 mg · m-3 for 5 min in a flow-past header,while the normal control group were PMVECs in quiescent condition. The supernatants and lysates of PMVECs were harvested at 0.5,1,2,4 and 8 h,respec?tively, after execution. The contents of TNF-α,IL-1β,ICAM-1,MMP-2 and MMP-9 were measured by ELISA,and the activity of MMP-2 and MMP-9 was measured by gelatin zymography. RESULTS① According to the morphologic characteristics of cell growth and the expression of specificity antigens and the bind experiment of phytohemagglutinin,the cells separated and purified by modified method shared the characteristics of PMVECs.②TNF-αwas rapidly expressed by PMVECs at 0.5 h post PFIB stimulation and the maximum value was achieved at 2 h post PFIB stimulation(P<0.05). The newly synthesized TNF-α was slowly released out of the cells. The maximum TNF-α in the supernatant was achieved at 4 h post stimulation.③Within 2 h of stimulation,PMVECs synthesized a large amount of IL-1β and peaks at 2 h. However,IL-1βwas never released to the extracellular milieu.④The amount of ICAM-1 was rapidly synthesized by PMVECs after PFIB stimulation,but at a low level.⑤After stimulation with PFIB,MMP-2 in the supernatant of PMVECs culture was gradually increased,peaked at 2 h and then decreased subsequently. The biological activity of MMP-2 in the supernatant was also enhanced after PFIB stimulation. PFIB did not stimulate synthesis or secretion of MMP-9,indicating that PMVECs were not the main source of MMP-9 during PFIB inhalation-induced acute lung injury. CONCLUSION PFIB stimulates the surviving PMVECs to synthesize a large amount of TNF-α,IL-1β, MMP-2 and conjunctive ICAM-1.

OBJECTIVE To investigate the inhibitory effect of lead acetate on transient receptor potential A1(TRPA1)channel. METHODS TRPA1-mediated calcium influx in mice dorsal root ganglion(DRG) neurons and HEK293 cells expressing nouse TRP1 (mTRPA1) and human TRPA1 (hTRPA1) was recorded by intracellular calcium imaging. TRPA1-mediated currents were detected by two-electrode voltage clamp. RESULTS Lead acetate 3.0 and 10.0μmol·L-1 inhibited external calcium influx in DRG neurons by(36.7 ± 4.1)% and(79.4 ± 3.1)%(n=5),respectively. The inhibitory effect of lead acetate on hTRPA1-mediated current was concentration-dependent. Lead acetate 0.3, 1.0, 3.0, 10.0 and 30.0μmol · L-1 inhibited the amplitudes of currents by(1.0 ± 0.7)%,(11.6 ± 0.8)%,(57.7 ± 3.2)%,(93.6 ± 2.6)%and(91.2±2.0)%(n≥4),respectively,with the IC50 2.4μmol·L-1. CONCLUSION TRPA1 channel may be an endogenous target of lead. Lead acetate inhibits TRPA1 channel at a very low concentration.

OBJECTIVE To investigate the delayed cytotoxicity effect of chlorpyrifos (CPF) with?drawal on primary hippocampal neurons. METHODS Hippocampal neurons were prepared from SD rat fetuses on the 17th day of gestation. Seven days after culture,neurons were exposed to CPF 10 and 30 μmol · L-1,respectively,for 72 h or for 48 h followed by CPF withdrawal for 24 h. CCK-8 kit and neuronal nuclei(NeuN), 5-bromodeoxyuridine(BrdU) and β Ⅲ tubulin immunofluorescence expression methods were used to evaluate the cell viability. RESULTS Compared with normal control, no significant cytotoxicity was found after CPF 72 h continuous exposure. However,CPF 48 h expo?sure followed by CPF withdrawal for 24 h induced evident cytotoxicity. The amount of BrdU positive and β Ⅲ tubulin positive hippocampal neurons were both decreased significantly(P<0.05),and cell survival and viability reduced after CPF withdrawal. CONCLUSION CPF exposure withdrawal can induce more seriously delayed cytotoxicity than continuous exposure in rat primary hippocampal neurons.

OBJECTIVE To investigate the effect of new baicalin(BC) metal ions(Co2+,Cu2+, and Ni2+)complexes(BMCs)on ion channels Kv1.4 and Cav3.2. METHODS HEK293 or CHO cells loaded with various ion channels(hERG,Kv1.2,Kv1.3,Kv1.4,Kv1.5,Kv1.6,Kv1.7,Kv1.8,Kir1.1, Kir2.1,KCNQ and Cav3.2)were obtained by stable transfection method. Whole-cell patch-clamp tech?nique was used to record current changes of each ion channel induced by BC and BMC in 10μmoL · L-1. The effect of different concentrations(0.3,1,3,10 and 30μmoL · L-1)of BC-Co and BC-Cu on Kv1.4 and Cav3.2 current was detected by whole-cell patch-clamp technique. RESULTS A model of HEK293 cells or CHO cells that stably expressed various ion channels was obtained. BMCs (BC-Co,BC-Cu and BC-Ni)had some impact on various ion channels,especially on Kv1.4 and Cav3.2. The inhibitory rate induced by BC-Co,BC-Cu and BC-Ni(10 μmol · L-1)was 91%,76% and-10%,respectively,for Kv1.4;and 43%,57%and-14%,respectively,for Cav3.2. IC50 of BC-Co was 1.69 and 0.81μmoL·L-1 for Kv1.4 and Cav3.2. IC50 of BC-Cu was 1.66 and 0.58μmoL · L-1 for Kv1.4 and Cav3.2. CONCLUSION BC-Cu and BC-Co concentration-dependently inhibit Kv1.4 and Cav3.2 ion channels.

OBJECTIVE To investigate the effect and underlying mechanism of Gypsophila elegans isoorientin on the proliferation and apoptosis of human HepG2 cells. METHODS HepG2 cells were treated with isoorientin 5,10,20,40,80 and 160μmol?L-1 for 24,48 and 72 h,respectively. Cell survival was analyzed by MTT assay. HepG2 cells were treated with isoorientin 5,10 and 20μmol?L-1 for 48 h before the lactate dehydrogenase(LDH)level was detected. After treatment with isoorientin for 24 h, the variation of reactive oxygen species (ROS) was monitored by a fluorescence probe H2DCF-DA. HepG2 apoptosis and mitochondria membrane potential(MMP)were evaluated by flow cytometry. The activi?ties of caspase 3,8 and 9 were determined by colorimetry. The mRNA expression of Bcl-2 and Bax was determined by RT-PCR,and the protein expression of Bcl-2, Bax and cytochrome c was detected by Western blotting. RESULTS Isoorientin(5-160μmol?L-1)inhibited HepG2 cell survial in a concen?tration-dependent manner,the 50%inhibitory concentration(IC50)was 62.7±9.1,47.2±11.4 and(18.2± 7.5)μmol?L-1 after treatment with isoorientin for 24,48 and 72 h,respectively. Compared with the cell control group,treatment with isoorientin 5,10 and 20μmol?L-1 significantly increased the LDH level and cell apoptosis rate(P<0.05). Moreover,isoorientin 10 and 20μmol?L-1 notably increased the production of ROS,decreased the MMP(P<0.05),and increased the activities of caspase 3 and 9. RT-PCR analysis and Western blotting showed that isoorientin significantly decreased the mRNA and protein expressions of Bcl-2,decreased the mRNA and protein expressions of Bax(P<0.05),and inhibited the protein expression of cytochrome c(P<0.05). CONCLUSION Isoorientin inhibits HepG2 cell prolif?eration,but promotes cell apoptosis,which is closely related to the regulation of the mitochondrial apoptosis pathway.