Chemotherapy is one of the three most common treatment modalities for cancer .However , chemotherapy as current firstline therapy induces significant side effects and limited efficacy ,leading to multidrug resistance and fast recurrence challenging the patient survival rate .Drug metabolizing enzymes ( DMEs) and efflux transporters promote the metabolism,clearance,and detoxification of chemotherapeutic agents .Nuclear receptors, especially pregnane X receptor (PXR,NR112)and constitutive androstane activated receptor (CAR,NR113),reg-ulate the expressions of target genes that could encode phase I DMEs ,phase II DMEs,and efflux transporters in the development of multidrug resistance ( MDR) during chemotherapy .Recent studies have revealed that PXR and CAR play pivotal roles in MDR of various human carcinomas .And their expressve levels or activation statuses could predict the risk of drug resistance in the patients subjected to chemotherapy .Accordingly ,PXR/CAR antag-onists,combining with existing chemotherapeutics that activate PXR /CAR,are promising options that could over-come MDR in cancer.

RNA interference is an evolutionarily conserved surveillance mechanism that responds to double-stranded RNA by sequence-specific silencing of homologous genes. It can block and regulate gene expression. Over the past few years, RNAi has already begun to revolutionize experimental biology in organisms ranging from unicellular protozoans to mammals. In neuroscience, we are only begining to know the mechanistic complexity of this process and its biological ramifications.

Aim To evaluate the effects of fibrinolytic enzyme FⅡ from agkistrodon acutus venom on an experimental model of kidney thrombus induced by lipopolysaccharide(LPS). Methods The model of microvascular thrombosis in the rabbits kidney was performed by the method of Hermida, which was induced by infusing LPS. Treatments were begun simultaneously with LPS infusion, through the contralateral marginal ear vein. Six different groups were established: NS 10 ml?h~-1 was infused as the negative control group, urokinase ~20 000 IU?kg~-1 ?h~-1 as positive control group, FⅡwas infused with the dosage of 0.1(Low-dose), 0.3 (medium-dose),0.6 (high-dose) mg?kg~-1 ?h~-1 . The further rabbits, which were given neither LPS nor FⅡ, were infused with saline solution through both marginal ear veins. Kinney sections were examined for the presence of fibrin microthrombi. The measurement of FDP concentrations was used to assess the degradation of microvascular thrombosis. Results Intense fibrin deposition was also detected and FDP concentrations were (78.21?4.79)% and (84.27?6.21)% at 2 and 6 hours after LPS administration in LPS-control group. Little fibrin deposition was detected and FDP concentration also increased in urokinase control group. A lot of fibrin deposition was detected in Low-dose FⅡ group,little fibrin deposition was detected in medium-dose FⅡ group, and no fibrin deposition was detected in high-dose FⅡ group. Additional all doses of FⅡ led to a significant increase in FDP concentration as compared with LPS-control group (P

Aim To find out the relationship between muscarinic receptor and reactive oxygen species (ROS) and the probable differences between the four muscarinic receptor subtypes. Methods We transfected the plasmid encoding muscarinic receptor (including subtypes: M_1, M_2, M_3 and M_4) into PC12 cells. Then PC12 cells were exposed to hydrogen peroxide (H_2O_2), carbachol and other inhibitors such as atropine, LY294002 and PD98059. Results The results showed that activation of muscarinic receptor by carbachol protected PC12-M_1, PC12-M_2,PC12-M_3 and PC12-M_4 cells from apoptosis induced by H_2O_2. There was no statistical difference in the protective effect between these four muscarinic receptor subtypes. By using the inhibitors, we found that atropine and LY294002 blocked the protective effect of activation of muscarinic receptor on apoptosis induced by H_2O_2. Conclusion Activation of muscarinic receptor retarded the apoptosis induced by H_2O_2. There was no difference between the four muscarinic receptor subtypes. The protective effect was mainly mediated by the activation of muscarinic receptor and phosphatidylinositol-3 kinase (PI3K).

AIM To investigate the protection of plicamycin on apoptosis in cerebellar granule neurons (CGN) of rat. METHODS TUNEL, Hoechst 33258 staining, agarose gel electrophoresis and fluorescein diacetate staining were used to detect morphological and biochemical characteristics of apoptosis in primary rat CGN. RESULTS Being pre-incubated with plicamycin for 1 h and lasting for 24 h, rat CGN apoptosis induced by low potassium basal modified Eagle′s medium for 24 h was inhibited in a plicamycin concentration-dependent manner. This effective concentrations of plicamycin were from 50 to 200 nmol·L-1, and the maximum inhibitory rate of plicamycin on CGN apoptosis was near 80% at 200 nmol·L-1. CONCLUSIONPlicamycin inhibits rat CGN apoptosis induced by low potassium.

AIM To construct a non-normalized cDNA library from Agkistrodon acutus venom gland as an imtial step to develop new and more effective venom by genetic engineering technique for screening and expressing target genes. METHODS The total RNA was extracted from fresh venom gland using Trizol. mRNA was reversely transcripted to cDNA using superscriptⅡ reverse transcriptase. Second-strand synthesis was performed using DNA polymeraseⅠ. After adding EcoRⅠ adaptor, phosphorylating the end and digesting with XhoⅠ, the cDNA was collected in five fractions (<0.25 kb, 0.25-0.5 kb, 0.5-1 kb, 1-2 kb and >2 kb) using the QIAquick Gel Extraction kit and ligated to pBluescriptⅡ vectors. The five libraries obtained were plated by infecting E.coli DH10B, constructing a cDNA library of Agkistrodon acutus venom gland. Sequencing clones at random, 8696 high quality 5′ end expressed sequenced tags (ESTs) were obtained and analyzed. The initial sequences were assembled into 2855 clusters. Among which, one of the clusters (Agkihagin) consisting of 74 ESTs was identified as a novel metalloprtoteinase based on RT-PCR and sequence analysis. RESULTSThe titers of library were 2.048×106. The novel metalloproteinase belonged to PⅢ type metalloproteinase. Its open reading frame was composed of 1827 nucleotides and coded a pre-zymogen of 608 amino acid with zinc-binding domain for metalloproteinase and Asp-Glu-Cys-Asp(DECD) domain for disintegrin. CONCLUSION The capacity of cDNA library of venom gland is above the general level of cDNA library. It would be a helpful platform to construct a catalog for transcripts in the venom gland of the Agkistrodon acutus. The sequence analysis indicates that the deduced amino acid sequence of the identified gene for metalloproteinase share the highest 87% identity with the metalloproteinase genes of other snakes in the GenBank. It lays a good foundation for the study of structure-function relationships of snake venom metalloproteinases.

AIM: To Screen and identify differentially expressed genes that involved in apoptosis model in rat cerebellar granule neurons (CGNs).METHODS: The rat cerebellar granule neurons were isolated and primarily cultured. Fluorescent differential display RT-PCR (FDD RT-PCR) was performed to screen differentially expressed ESTs in the apoptosis model of primarily cultured rat CGNs. ESTs were subcloned into pGEM-T EasyTM vector and then sequenced. Alignment assay in non-redunant database was applied for encoding information. Reverse Northern blotting was used to appraise the results from DDRT-PCR.RESULTS: 164 pieces of differentially expressed ESTs were obtained by FDDRT-PCR. 17 of them were subcloned and sequenced. 5 ESTs of 17 were confirmed to be positive results by reverse Northern blotting. CONCLUSION: DD-PCR is a rapid, simple-operation and sensitive method for screening differentially expressed genes, which would contribute to the molecular mechanisms of apoptosis/survive of CGNs.

AIM: To analyze the subcellular localization of Arnt2 in rat cerebellar granule neurons (CGNs). METHODS: Based on the amino acids sequence of Arnt2 (LOCUS:NP_036913), the subcellular localization of Arnt2 in eukaryotic cells and the nuclear export signals (NES) of Arnt2 were predicted in CBS bioinformatics database. The subcellular localization of Arnt2 in rat cerebellar granule neurons was detected by the method of laser scanning confocal microscopy (LSM) analysis. RESULTS: It was predicted that Arnt2 located in nuclei of eukaryotic cells with the most probability, while located in cytoplasmic mitochondria with a slight possibility. A nuclear export signal was found in Arnt2 amino acids sequence, it was identified to be the leucine of No.143 that located in N-terminal of Arnt2 amino acids sequence. Finally, the result of LSM analysis shows nuclear localization of Arnt2 in rat CGNs. CONCLUSION: Arnt2 is located in nuclei of normal rat CGNs, it suggests that Arnt2 has the tendency to translocate into mitochondria after induced by some of inducible factors, for both the possibility of mitochondria localization and NES exist in Arnt2 amino acids sequence.

【Objective】To investigate the molecular mechanism of dopamine (DA)-induced apoptosis in cultured cerebellar granule neurons (CGNs) and the effect of muscarinic cholinergic receptor (mAChR) agonist carbachol on it.【Methods】The apoptosis of neurons was measured by phase-contrast microscopy,Hoechst 33258 nucleus staining and DNA fragmentation agarose gel electrophoresis.The neuronal viability was measured by fluorescein diacetate (FDA) staining.The activation of extracellular signal-regulated protein kinase (ERK) was determined by Western blot.【Results】Dopamine increases the phosphorylation of ERK and induces apoptosis in CGNs,which is blocked by both carbachol and PD 98059.The protective effect and the inhibiting ERK phosphorylation of carbachol were blocked by atropine.【Conclusion】DA-induced apoptosis in CGNs may be mediated by activation of ERK.Carbachol protects CGNs from DA-induced apoptosis by activating mAChR and subsequent inhibition of activation of ERK.

AIM: To observe the short-term curative effect of Suxiao Jiuxin Pill on coronary heart disease (CHD) and angina pectoris. METHODS: 187 patients were divided into two groups at random:the treatment group with 105 cases and the control group with 82 cases. The treatment group was given Suxiao Jiuxin Pill 6 pills tid and the control group was given Compound Danshen Dripping Pill 6 pills tid orally. The treating period was 2 weeks. Before and after treatment angina pectoris was observed and electrocardiogram was determined. RESULTS: Suxiao Jiuxin Pill could significantly alleviate syndrome of CHD in patients, lessen electrocardiogram and decrease dosage of nitroglycerin. The total effective rate on angina pectoris and the effective rate in improving electrocardiogram was higher in treatment group than that in control group (P