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.

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.

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 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

BACKGROUND:Short-segment pedicle screw technology has been extensively used in the treatment and repair of thoracolumbar burst fractures in the clinic. However, it requires further investigations whether bone graft through unilateral pedicle channel has superiority. ＜br> OBJECTIVE:To observe the perioperative effects on thoracolumbar burst fractures posterior by bone graft through unilateral pedicle channel with short pedicle screw segment fixation. ＜br> METHODS:Clinical data of 57 cases of thoracolumbar burst fractures without neurological symptoms treated by short segment pedicle screw internal fixation therapeutic method at the Department of Orthopedics, The First People’s Hospital of Yinchuan City from January 2009 to December 2013 were retrospectively analyzed. According to therapeutic methods, they were divided into three groups:pedicle screw fixation through injured vertebrae (n=21), unilateral pedicle screw fixation with vertebrae pedicle screw (n=19), and the short segment pedicle screw fixation with unilateral pedicle screw fixation and bone graft through pedicle channel (n=17). We observed the changes in operation time, the blood loss volume in operation, postoperative drainage blood (at the period of removing the drainage tube), injured vertebrae height’s ratio, the Cobb’s angel of injured vertebrae sagittal plane, vertebral canal encroachment rate and visual analog scale. ＜br> RESULTS AND CONCLUSION:(1) The unilateral pedicle screw fixation and bone graft by pedicle channel could prolong operation time and increase postoperative drainage blood volume (P＜0.05). (2) Three operation methods improved vertebral body height. (3)There were no significant differences in Cobb’s angle postoperation among the three groups (P>0.05). These data suggested that the bone graft through pedicle channel was not the factors to improve the spine kyphosis. (4) Bone graft by pedical channel improved vertebrae channel value. (5) Bone graft through pedicle channel combined with screw implantation could improve pain after treatment. These findings indicated that the therapeutic technology of short pedicle screw fixation on thoracolumbar vertebral burst fracture with unilateral pedicle fixation and bone graft through pedicle channel is a safe reliable operation method.

【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

[Objective] To purify and characterize a novel factor X activator,Fve-1 from Daboia russelli siamensis (Myanmar) venom.[ Methods]F V e-1 was purified by ion-exchange chromatography and gel filtration.The hemostatic activity of F V e-1 was determined based on chromogenic substrates.The fibrinogen-clotting activity of F V e-1 was also determined.Thermal stability, pH stability,enzyme activity,and inhibition of F V e- 1 were determined by its remaining procoagulant activity.N-treminal sequence was determined by the method of automated Edman degradation.[ Results ]F V e-1 was achieved by chromatography with a molecular weight of 13,808 and an isoelectric point of 4.6. The hemostatic activity of 0.5 mg Fve-1 was equal to that of 1.5625 u thrombin or that of 54.93 ng RVV X. F V e-1 primarily activated F X, but did not affect on prothrombin and fibrinogen. The suitable pH and temperature range of F V e-1 was 6.5-7.5 and 25-60 ℃,respectively.The activity of F V e-1 was enhanced by Ca2+ and inhibited by EDTA and DTT.The N-terminal sequence of F V e-1 was NH2-N-L-Y-Q-F-G-E-M-I-N.[Conclusion] F V e-1 is a factor X-activating enzyme,which could activate FX to FX a,but have minimal effect on prothrombin and fibrinogen.

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.