Objective To construct a lentiviral vector over-expressing β-catenin gene by multisite Gateway technology and confirm its effect.Methods By using multisite Gateway clone technique,the entry clone of pDown-Ctnnb1 was constructed using BP recombination reaction.Then,LR recombination reaction was performed among pUp-EF1A,pDown-Ctnnb1,pTail-IRES/DsRed-Express2 and pLV.Des3d.P/puro to generate an expression clone of pLV.EX3d.P/puro-EF1A＞Ctnnb1 ＞IRES/DsRed-Express2.In each step,PCR and sequencing analysis were used to verify the constructions.When it was verified that plasmids were transfected into 293T cells,PT-PCR was performed to determine the mRNA level of β-catenin gene.Results Both PCR and sequencing analysis revealed that β-catenin over-expression gene was inserted into the target site and the insertion sequence was perfectly corrected.The RT-PCR results showed that the expression of β-catenin gene was significantly upregulated.Conclusion The lenvivirus-mediate β-catenin over-expression gene was successfully constructed..

OBJECTIVE To discuss the best scheme for fungemia detection by analyzing and comparing the ability and performance characteristics of the BACTEC 9120 automated blood culture system and 4 kinds of blood culture bottles in the detection of simulated fungemia.METHODS Simulated blood culture was produced using 65 fungi isolates from clinical specimens and BACTEC Plus Aerobic/F,Plus Anaerobic/F,Peds Plus/F and Myco/F Lytic blood culture bottles and detected by BACTEC 9120 automated blood culture system.The final inoculum densities were 1-5 CFU/ml blood.The(time to detection TTD) of simulated blood culture with different concentrations of suspension produced using 2 kinds of standard strains and 4 kinds of blood culture bottles was compared.RESULTS From the 260 bottles in this study 216 had growth detected by the BACTEC 9120 blood culture system.The positive rates of BACTEC Plus Aerobic/F and Anaerobic/F,which were 90.77%and 41.54%,respectively,were significantly(P

AIM: To determine proteins molecular weights in seed of Cortex Magnolia and identify its varieties by gel ecectrophorsis. METHODS: SDS-PAGE technique. RESULTS: There are distinct distinguish in SDS-PAGE. The molecular weights of the major proteins of the samples are analyzed by it. CONCLUSION: Cortex Magnolia can be discerned by gel electrophorsis and the molecular weights of them.

OBJECTIVE:To evaluate the bioequivalence of domestic secnidazole tablet,capsule and imported secnidazole tablet in healthy volunteers.METHODS:18healthy volunteers were randomly divided into3groups according to a triple-cross design,all the volunteers were given a single dose of1g secnidazole,the interval for washout period of3times adminis?tration was14days.The plasma drug concentration of secnidazole was determined by HPLC-UV.RESULTS:The main pharmacokinetic parameters of homemade tablet and homemade capsule and imported tablet were as follow,t max were(2.00?1.93),(2.67?2.14)and(1.54?1.53)h respectively,t 1/2 were(28.56?4.98)、(29.69?6.81)and(27.16?5.06)h,C max were(25.50?2.74),(24.27?3.76)and(25.64?4.10)?g/ml respectively.AUC 0～t were(736.03?73.20),(704.78?88.51)and(737.77?76.02)(?g?h)/ml respcetively.AUC 0～∞ were(886.36?114.50),(864.57?172.27)and(870.64?100.21)(?g?h)/ml respectively.The relative bioavailability of homemade tablet was(100.02?6.73)%,and that of homemade capsule was(95.91?10.66)%.CONCLUSION:3preparations of secnidazole are bioequivalent.

Objective To observe the effects of exercise on serum adiponectin and adiponectin receptor (AdipoR) level in skeletal muscle of insulin-resistant rats. Methods A total of 30 healthy male rats were randomly divided into a control group ( NC, n = 8) and a high-fat group ( HF, n = 22), fed with normal chow and high fat diet, respectively. Eighteen weeks later, the high-fat group was randomly divided into a high-fat diet control group (HC, n = 10) and an exercise group (HE, n = 12). The HC and HE group were continually fed with high fat diet, while the HE group was administered with swimming training for 6 weeks in addition at the same time. After 24 weeks, the insulin sensitivity index was calculated, and serum adiponectin level was detected by using ELISA. The expressions of AdipoR mRNA in skeletal muscle were detected with real-time fluorescence quantitative polymerase chain reaction (PCR). Results After 18 weeks, compared to NC group, the insulin sensitivity index of HF group decreased significantly. It suggested that insulin resistance appeared in HF group. Twenty-four weeks later, compared to NC group, the ISI of HC group was significantly decreased, meanwhile the level of serum adiponectin, expression of AdipoR1 and AdipoR2 mRNA in skeletal muscle of HC group were 71.9% , 59.9% and 69.2% of those of the NC group, respectively; compared to HC group, the ISI was increased significantly by exercise, meanwhile the expression of AdipoR1 mRNA in skeletal muscle was significantly increased by 1.33 times, however the level of serum adiponectin and the expression of AdipoR2 mRNA in skeletal muscle were not altered in HE group. Conclusion Six weeks of exercise improves insulin sensitivity through increasing the expression of AdipoRI mRNA in skeletal muscle.

Objective To investigate the effect of intermittent pneumatic pressure pump on the formation of deep venous thrombosis of lower extremity in elderly patients undergoing thoracotomy. Methods Fifty patients undergoing thoracic surgery were managed with intermittent pressure inflation pump besides general preventive measures. The pre-and postoperative blood flow in the lower extremity and incidences of deep venous thrombosis were compared. Result The differences in blood flow in the lower extremity and incidences of deep venous thrombosis were not statistically significant (P>0.05). Conclusion Intermittent pneumatic pressure pump can promote pulsatile blood flow in the lower extremity so as to effectively prevent formation of deep venous thrombosis in elderly patients after thoracotomy.

Neuron is the basic unit of the biological neural system. The Hodgkin-Huxley (HH) model is one of the most realistic neuron models on the electrophysiological characteristic description of neuron. Hardware implementation of neuron could provide new research ideas to clinical treatment of spinal cord injury, bionics and artificial intelligence. Based on the HH model neuron and the DSP Builder technology, in the present study, a single HH model neuron hardware implementation was completed in Field Programmable Gate Array (FPGA). The neuron implemented in FPGA was stimulated by different types of current, the action potential response characteristics were analyzed, and the correlation coefficient between numerical simulation result and hardware implementation result were calculated. The results showed that neuronal action potential response of FPGA was highly consistent with numerical simulation result. This work lays the foundation for hardware implementation of neural network.
Action Potentials ; Computer Simulation ; Humans ; Models, Neurological ; Neural Networks (Computer) ; Neurons ; cytology ; Synaptic Transmission

Objective To study the immune system regulatory effects of CD8+CD28-regulatory T lymphocytes in an experimental bone marrow mesenchymal stem cell treatment of autoimmune encephalomyelitis.Methods A model of autoimmune encephalomyelitis (EAE) was established in twenty-eight C57BL/6 female mice,6 to 8 weeks old weighing 16 to 20 g using myelinoligodendrocyte glycoprotein 35-55 amino acid peptide (MOG35-55).The mice were randomly divided into a phosphate-buffered solution (PBS) group (n =7),an MSCs-Low group [n =7 which received an injection of 2× 105 mesenchymal stem cells (MSCs)],an MSCs-Med group (n=7,1× 106 MSCs),and an MSCs-High group (n=7,5×106 MSCs).Their clinical condition was then evaluated daily.On the 15th day after the MOG35-55 immunization,the different MSC doses were administered intravenously via the tail.On the 30th day the mice were sacrificed to observe any neuropathology of the spinal cord.At the same time,FACS flow cytometry was used to assay CD8+CD28-T cells in the spleen.Results Compared with the PBS control group,the MSC treatment effectively alleviated illness among the mice by the 15th day after the immunization.The maximum and average disease scores and clinical illness scores had all improved significantly.The medium dosage worked best.Neuropathological analysis showed that the MSC treatment could significantly reduce the invasion of inflammatory cells and minimize demyelination in the spinal cord.Furthermore,the CD8+ CD28-regulatory T cells in the spleens of the MSCtreated animals increased compared with the PBS control group,though the secreted levels of IL-10 showed no obvious difference.Conclusions Treatment with MCSs can promote the recovery of neural function in autoimmune encephalomyelitis,at least in mice.CD8+CD28-regulatory T cells may not be the main effector cells,playing only a synergistic therapeutic role.

To get the information about civil attitudes and main affecting factors of cadaver donation in Zhengzhou.Make stochastic visits to part of dwellers in Zhengzhou urban districts and then study the statistical data.The main affecting factors are traditional ideas and unsound legal system.So,in order to have a good effect,we should strengthen the dissemination of cadaver donation,renovate ideas,and strengthen relevant legislation for cadaver donation.

Objective: To investigate the pharmacodynamics of modified Xiaochaihutang granules (MXG) and its active ingredients group (AIG) on gastric ulcer in order to provide scientific basis for the clinical effect and lay foundation for the development of related hospital preparations.Methods: Gastric ulcer models induced by pyloric ligation,acetic acid and ethanol were applied to observe the curative effects of MXG and its AIG, and the effects of MXG and its AIG on gastric secretion, gastric acid content, pepsin activity, superoxide dismutase (SOD) and malondialdehyde(MDA) contents were also studied.Results: Compared to the model control group,MXG and AIG could significantly reduce the ulcer index(P<0.01), total acidity(P<0.01),and pepsin activity (P<0.05 or 0.01), increase the SOD activity (P<0.05)and inhibit the increase of lipid peroxide MDA.Conclusion: MXG and AIG show better control effects on acute and chronic gastric ulcer, and the effects are related to reducing the activity of pepsin, increasing the SOD activity, inhibiting the increase of lipid peroxide MDA and so on.