AIM To examine antithrombotic effects of arecoline on the arterial thrombosis induced by carrageenin in mice through modulating the functions of endothelium and determine its mechanisms from hemostatic system, the platelet aggregative functions and the bioactive factors released by vascular endothelial cells. METHODS Kappa carrageenin was given ip in mice and mice were fed at the temperature of 20 to 21 degrees and at the humidity of 30 percent to 50 percent. RESULTS On the foregoing models of thrombosis, arecoline could antagonize the formation of thrombosis through activating the endothelial target for acetylcholine in a dose dependent manner and its antithrombotic potency was 250 to 500 times greater than aspirin; while under the same conditions, pilocarpine could not antagonize the formation of thrombosis. The levels of TT, PT, KPTT and MAR had no prominent changes compaired with control groups. The levels of t-PA became higher greatly than normal and the levels of PAI 1 became lower greatly than normal 2 hours after intravenous injection of arecoline in rats. Arecoline could decrease the higher plasma levels of thromboxane A2 and increase the lower plasma levels of prostacyclin in a dose dependent manner in the mice tail thrombosis induced by carrageenin. CONCLUSION The antithrombotic effects of arecoline are associated with activating the endothelial target for acetylcholine closely, but are not associated with muscarinic receptors,and not relevant to hemostatic systems or functions of platelet aggregation directly.

OBJECTIVE To develop a method to construct three dimensional finite element model of nasomaxillary complex,in order to provide the foundation of studying nasomaxillary biomechanics characteristics.METHODS DICOM data from HelixCT were analyzed and reconstructed in software Mimics,then complete constructing three dimensional finite element model of nasomaxillary complex in Ansys.RESULTS The nasomaxillary complex model was similar to original case in structure,which is a high finite element model.CONCLUSION Computer assistant in constructing finite element model from DICOM data is a convenient,exact and efficient method .The nasomaxillay complex finite element model is exact and reproductable,and can provide the foundation of studying biomechanics mechanism of nasomaxillary fracture in otolaryngology.

Objective:To establish a rapid method to detect drug-resistance genotypes of extended spectrum-β-Lactamases (ESBLs) produced by gram negative bacillus using the real -time fluorescence quantitative PCR. Methods: According to clinical common genotypes of ESBLs, SHV, TEM.CTX-M.OXA and their homology, 9 pairs of specific primers were designed including SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1, OXA-2 and OXA-10. To extract DNA template by boiling assay, and then establish and grade up SYBR GREEN I real-time fluorescence quantitative PCR reaction system, finally definite real-time fluorescence quantitative PCR method. Its precision and range of linearity were tested. With established assay 51 multi- drug resistant ESBLs- E. coli K. pneumoniae were detected and compared with improved three dimensional extract tests. Results: Except OXA-2, 8 genotypes SHV, TEM, CTX-M-1, CTX-M-2, CTX-M-8, CTX-M-9, OXA-1 and OXA-10 were amplified by quantitative PCR from 39 ESBLs+ and 51 multi-drug resistant ESBLs-E. coli K. pneumoniae and confirmed by sequence testing. The range of linearity was 3×10~3-3×10~8 copies/mL, r =-0.994 7. Repetitive experiments showed that the average coefficient of variation between -runs was 9.6%. Comparing with three dimensional extract test, there was no significant difference (χ2 = 1.125,P> 0.05). Conclusion: Testing drug-resistance genotypes of ESBLs with SYBR GREEN I real-time fluorescence quantitative PCR is a rapid,specific and sensitive method, which is capable of inspecting genotypes of ESBLs from clinical strains.

Objective To investigate the psychological health states of practical nurse and to effectively conduct clinical psychological guidance.Methods 368 practical nurse were determined by SCL-90 and compared with normal control.Results There were significant differences between practical nurse and normal control,such as compulsion,sensitive interpersonal,depression,anxiety,terror crankiness(P<0.05～P<0.01),and no remarkable difference in other factors(P>0.05).Conclusion Nurse's gross mental health level were worse than normal control,especially in compulsion,sensitive interpersonal,depression,anxiety,terror crankiness. So, mental health education should be enhanced for them.

Objective To increase the quality of blood transfusion medical record and strengthen the management of clinical blood transfusion by establishing a management system for clinical blood transfusion.Methods The management system of clinical blood transfusion was developed by using Sybase PowerBuilder 10.5 program and Oracle 8/8i database,through the function module's development of blood application and evaluation by using C/S structure.Results The management system of clinical blood transfusion realized the exchange of the internal data information with the blood information management system and LIS database,and implemented online audit of transfusion application and evaluation,which improved the work efficiency and reduced the human error.Conclusion The management system of clinical blood transfusion can improve the quality of blood transfusion medical record and realize real-time regulation of clinical blood transfusion to ensure the safety of transfusion.

Objective To establish a rapid specific quantitative assay for Bacillus anthracis detection. Methods According to the principle of complex probe quantitative assay, the primers and quantitative probes targeted at chromatosome DNA rpoB were designed and applied to detect Bacillus anthracis. The influence factor of quantitative PCR were determined. Results The optimal system of this method was aquired: the length of quenching probe is 15mer,the ratio of fluorescent probe to quenching probe is 1/2 and the concentrtion of Mg 2+ is 3 mmol/L.The sensitivity of this assay for Bacillus anthracis is 10 3 copies. It can distinguish Bacillus anthracis from other closely related Bacillus. Conclusion The method can rapidly quantitatively detect the Bacillus anthracis with high sensitivity and specificity, it can be applied to clinical diagnosis.

Objective To study the optimum ultra-dry method and moisture at different storage time for Salvia miltiorrhiza seeds and find the principle of storability.Methods S.miltiorrhiza seeds were dried by silica gel at room temperature and by the oven at constant temperature 50 ℃ to obtain various moisture content before stored sealed at room temperature.The optimum ultra-dry method and the optimal moisture were evaluated by measuring the germination rate,germination tendency,and vigor index,etc.Soluble sugar and MDA content were measured to investigate the seed storability.Results Desiccation by silica gel was more proper than by oven;ultra-dry storage of seeds has obvious advantages at the early stage,but with the prolong of the storage time,the advantages decreased;The optimal moisture for S.miltiorrhiza seeds storage at room temperature is about 7.5%;Seed storability is closely related to soluble sugar content in the seeds.Conclusion S.miltiorrhiza seeds can be ultra-dry stored to preserve germplasm resources.

AIM: To investigate the protective effect of microRNA-218 (miR-218) silencing on kidney tissue of streptozotocin (STZ)-induced diabetic nephropathy rats and the potential mechanism.METHODS: The diabetic rat model was established by a single intraperitoneal injection of STZ (50 mg/kg).Meanwhile, the miR-218 short hairpin RNA (shRNA) lentiviral vector was constructed.The Sprague-Dawley rats were randomly divided into 4 groups: healthy control group, diabetes group, empty vector group and miR-218-shRNA group.The blood glucose, 24 h urinary protein, serum creatinine (SCr) and blood urea nitrogen (BUN) in the rats at different time points (4, 8 and 12 weeks) were measured by an automated analyzer.The expression of miR-218 was detected by RT-qPCR, while the expression of heme oxygenase-1 (HO-1), nephrin and p38 mitogen-activated protein kinase (p38 MAPK) at mRNA and protein levels in the kidney tissues was determined by RT-qPCR and Western blot.The caspase-3 activity was detected by caspase-3 activity assay kit, and the cell apoptosis of the kidney tissues was analyzed by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).RESULTS: Compared with healthy control group, the expression of miR-218 was significantly increased in STZ-treated rats.Meanwhile, the concentrations of blood glucose, 24 h urinary protein, SCr and BUN were significantly increased in STZ-treated rats (P<0.05).The mRNA and protein expression of HO-1 and nephrin was significantly decreased, while the level of phosphorylated p38 MAPK was significantly increased in STZ-treated rats.In addition, the activity of caspase-3 was also significantly increased in STZ-treated rats.When the model rats were infected with miR-218-shRNA, the expression of miR-218 was significantly decreased and the above effects were markedly reversed.Furthermore, TUNEL results showed that compared with diabetic group and empty vector group, miR-218 silencing significantly attenuated the cell apoptosis in the kidney tissues in miR-218-shRNA group.CONCLUSION: miR-218 is involved in the kidney injury in diabetic rats, and silencing of miR-218 by lentiviral vector-mediated miR-218-shRNA transfection effectively inhibits kidney cell apoptosis, suggesting that miR-218 is a potential target for the treatment of diabetic nephropathy.

Objective To investigate the specific anti-tumor immune response induced by dendritic cells (DCs) transfected with total RNA of human pancreatic cancer Capan-2 cells.Methods DCs were isolated and cultured from peripheral blood mononuclear cells (PBMCs) derived from six patients with pancreatic cancer.Total RNA of Capan-2 cells and MUC4 mRNA were transfected into DCs by electroporation.The survival rate of transfected DCs was determined by MTT method and the expression of MUC4 mRNA in DCs was detected by Western blotting.The activity of cytotoxic T lymphocyte cells (CTLs) induced by DCs transfected with total RNA of Capan-2 cells were evaluated by IFN-γ ELISA and the induction of specific CTL response to the killing effect on pancreatic cancer cell in vitro were measured by 51 Cr standard cytotoxicity test.Results The survival rate of DCs transfected with total RNA of Capan-2 cells (DC-Capan-2-total RNA) showed a decrease in a time dependent manner and the survival rate was reduced to 60.81％ after transfection for 96 h.The survival rate of MUC4 mRNA transfection DCs (DC-MUC4 mRNA) was stable at around 80％.The difference of DCs surviral rate between the two groups was statistically significant (P ＜ 0.05).The amount of MUC4 protein expression of DC-Capan-2-total RNA was significantly lower than that of DC-MUC4 mRNA (P ＜0.05).The quantity of CTL IFN-γ release induced by DC-Capan-2-total RNA was (89.34 ± 3.85)U/mL and the quantity of DC-MUC4 mRNA induced CTL IFN-γ release was (21.77 ± 21.77)U/ml There was statistically significant between the two groups (P ＜0.05).In addition,the specific CTLs induced by DC-Capan-2-total RNA could effectively identify and kill the HLA-A2+/MUC4+ Capan-2 and the HLA-A2+/MUC4-PANC 1 cells,and could not effectively identify and kill the HLA-A2 /MUC4-MiaPaCa-2 cells and the HLA-A2-/MUC4 + AsPC-1 cells.Conclusions A more pronounced CTL anti-tumor immune response can be induced by DCs transfected with total RNA of Capan-2 cells compared with a single tumor associated antigen,but it is limited by MHC class Ⅰ antigen presented.

Objective:To study the influence of all-trans retinoic acid (atRA)on craniomaxillofacial development of C57 mice. Methods:Pregnant C57BL mice were divided into 4 groups(n =5)at gestation day (GD)1 0.Mice in three atRA-induction groups were given atRA of 60,80 and 1 00 mg/kg,respectively.The mice in control group were given the equivalent volume of corn oil.All pregnant mice were sacrificed at GD1 9 and the embryos were collected.Stereo microscope was used to observe the craniomaxillofacial morphology.Standardized radiographs were taken and cephalometric analysis was performed.Results:The embryonic body length and body mass of control group surpassed those of 80 and 1 00 mg/kg atRA groups(P <0.05,P <0.01 ).atRA induced craniomaxillofacial malformations and maldevelopment.The mice induced by atRA exhibited a shorter mandibular body and more retrusive position of max-illary and mandibular(∠NAK and ∠NBD)when compared with their norm(P <0.01 ).Significant decrease in craniofacial length (Op-Rh)was observed in all atRA-induced groups(P <0.01 ).Decreases in cranial vault height(Fp-Os)and cranial vault length(Pa-Na)dimensions were observed in 80 and 1 00 mg/kg atRA groups(P <0.05,P <0.01 ).Conclusion:Exogenous atRA dose-depend-ently induces retardation of craniomaxillofacial morphology in embryo of C57BL mice by inhibition of the sagital and vertical dimension development of the bone.