Animals ; Flunarizine ; pharmacology ; Hippocampus ; drug effects ; physiopathology ; Male ; Neurons ; drug effects ; physiology ; Penicillins ; adverse effects ; Rats ; Rats, Wistar ; Seizures ; chemically induced ; physiopathology

Objective To study the expression of vesicular GABA transporter and vesicular glutamate transporter 1 in de‐pression .Methods Mice was divided into control group and defeat group stochastically .By social defeat model and social avoidance , the defeat group was divided into two groups:susceptible group and unsusceptible group .Synaptic proteins were extracted respec‐tively from the 3 groups .We detected the expression abundance of VGAT and VGLUT1 by Western blot .Results Compared with the control group ,in susceptible group ,the residence time in the contact area was significantly reduced ,and the residence time in the corner area was significantly increased ,with statistical difference(P<0 .05) .In the prefrontal cortex and hippocampus ,Com‐pared with the control group and the unsusceptible group ,the expression levels of VGLUT1 and VGAT were increased in the sus‐ceptible group(P<0 .05) .there were no statistically significant in VGLUT1 and VGAT leveles between control group and the un‐susceptible group(P>0 .05) .In the striatum ,although the expression levels of VGAT and VGLUT1 were increased in susceptible group ,but in unsusceptible group ,the expression of these proteins also increased significantly .Conclusion The prefrontal cortex and hippo‐campus excitability and inhibitory vesicle transport were changed in depression ,which may relate to the transcription disorder .

Objective To develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

To introduce the application of CBL teaching method on neuroanatomy teaching in undergraduate of con -tinue education school .Cases selection and reconstruction , designing problems , organization , implementation and standardized teaching were analyzed .Finally, we discussed the advantages and disadvantages of CBL teaching method.

Objective To investigate the mechanism of brain derived neurotrophic factor (BDNF) regulated by differ-ent isoforms of tyrosine kinase receptor B (TrkB) in epileptic hippocampal neurons. Methods Primary hippocampal neu-rons were cultured in vitro for 7 days, and divided into two groups, ALLN (calcineurin inhibitor) group and Anisomycin (trans-lation inhibitor) group. ALLN group included control group, control+BDNF group, epilepsy group, epilepsy+BDNF group, control+ALLN group, epilepsy+ALLN group and epilepsy+ALLN+BDNF group. Anisomycin group was sub-divided into con-trol group, control+BDNF group, epilepsy group, epilepsy+BDNF group, control+Anisomycin group, epilepsy+Anisomycin group and epilepsy+Anisomycin+BDNF group. The immunofluorescent technique was used to identificate the hippocampal neurons. Epileptiform discharges were detected by electrophysiological techniques. Western blot assay was used to deter-mine the protein expression of TrkB and phosphorylated TrkB (p-TrkB) in all cell groups. Results (1) In ALLN group, the gray value of p-TrkB/TrkB was higher in control+BDNF group compared with that of control group, the value was higher in epilepsy+BDNF group than that of epilepsy group but was lower than that of control+BDNF group. The gray value of p-TrkB/TrkB was lower in epilepsy+ALLN+BDNF group than that of epilepsy+BDNF group, but no significant difference compared with that of epilepsy+ALLN group. (2) In Anisomycin group:the gray value of p-TrkB/TrkB was higher in control+BDNF group than that of control group. The gray value of p-TrkB/TrkB was higher in epilepsy+BDNF group than that of epilepsy group, but which was lower than that of control+BDNF group. The gray value of p-TrkB/TrkB was higher in epilepsy+Aniso-mycin+BDNF group than that of epilepsy+BDNF group and epilepsy+Anisomycin group. Conclusion The decreased ex-pression of TrkB.T can improve the inhibition of BDNF/TrkB signaling, and BDNF can activate BDNF/TrkB signal pathway in epileptic hippocampal neurons. The increased TrkB.FL protein level by ALLN can’t improve the inhibition of BDNF/TrkB signal pathway.

Objective To study the effect of miR-204 on BDNF/TrkB signaling and pathogenesis on the neuron model of epilepsy. Methods Primary hippocampal neurons were cultured in vitro for 7 days, and were divided into control group, control+BDNF group, epilepsy group, epilepsy+BDNF group , control+miR204 group, epilepsy+miR204 group and ep-ilepsy+miR204+BDNF group. The epilepsy model of hippocampal neurons was established by being exposed to Mg2+free me-dia for 3 hours. The miR-204 lentivirus vector was constructed. The effect of miR-204 on BDNF/TrkB expression was detect-ed by immunohistochemistry, patch clamp and Western blot technique. Results Compared with the control group, the TrkB phosphorylation level was higher in control+BDNF group. The TrkB phosphorylation level was lower in epilepsy+BDNF group than that of control+BDNF group, but it was higher than that of epilepsy group. The TrkB phosphorylation level was higher in epilepsy+miR204+BDNF group than that of epilepsy+BDNF group and epilepsy+ miR204 group. Conclusion BDNF and miR-204 can improve the inhibitory condition of BDNF/TrkB signaling and may play an important role in alleviat-ing epilepsy disease.

Objective To prepare endo-beta-N-acetylglucosaminidase H (Endo-H) expressed in Pichia pastoris, and apply it to N-glycosylation analysis .Methods One complete gene was synthesized on the basis of the cDNA sequence encoding Streptomyces plicatus reported in GenBank .The gene was cloned into the expression vector pPIC 9.The expression vector pPIC9-Endo-H was transformed into P.pastoris(JC308).The expression products were induced by methanol , puri-fied by two-step chromatography , used to analyze the glycan structures of RNaseB by the DNA sequencer assisted fluoro-phore-assisted carbohydrate electrophoresis (DSA-FACE)methods, and finally compared with peptide-N-asparagine amidase F(PNGase F).Results This enzyme expressed in P.pastoris(JC308) had the ability to hydrolyze natural or denatured high-mannose type of oligosaccharide linked by β-1,4-glycosidic bonds , but not complex-type oligosaccharide .The result of DSA-FACE showed that carbohydrate chains of Man 5 GlcNAc-Man9 GlcNAc could be obtained when RNaseB was hydrolyzed by Endo-H, and that Man5 GlcNAc2-Man9 GlcNAc2 chains became available when RNaseB was hydrolyzed by PNGase F . Conclusion Endo-H expressed in P.pastoris has bioactivity which can be used to analyze N-glycosylation with the method of DSA-FACE.

Objective To explore a space craft precooling temperature at which excessive thermal stress on the crew member could be prevented or reduced in an overheated launch or reentry module. Method Five young male volunteers wearing a space suit participated in 25 tests at sea level.The space suit was either ventilated in a volume air flow rate of 100 L/min (STPD) with ambient air at temperatures (Ta) of 15℃,10℃,and 5℃,respectively,or not ventilated. Rectal (Tr),mean skin (Tsk) and mean body (Tb) temperatures were measured. Result At Ta 15℃,Tr decreased without significance (from 37.0±0.2℃to 36.7±0.3℃) in 120-min tests,whereas Tsk and Tb decreased significantly,and subjects had local cold strain whether the space suit was ventilated or not; while at Ta 10℃,Tr decreased from 37.0±0.3℃ to 36.3±0.3℃(P<0.05),subjects had a whole body cold strain,and both Tsk and Tb dropped continuously and significantly. Conclusion Ambient temperature 15℃,at which the thermal comfort states of crew was not significantly degraded,was acceptable after precooling in a space craft.

12E7 Antigen ; Aged ; Antigens, CD ; metabolism ; Cell Adhesion Molecules ; metabolism ; Diagnosis, Differential ; Giant Cell Tumors ; pathology ; Giant Cells ; pathology ; Humans ; Male ; Mesenchymoma ; pathology ; Mucin-1 ; metabolism ; Myositis Ossificans ; pathology ; Osteosarcoma ; metabolism ; pathology ; surgery ; Penile Neoplasms ; metabolism ; pathology ; surgery ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Vimentin ; metabolism

Objective To prepare and identify rabbit polyclonal antibody against embryonic liver fordrin 3(ELF3),and investigate the distribution of ELF3 in mice tissue.Methods ELF3 specific N-terminal peptide was synthesized,and conjugated to Keyhole limpet hemocyanin(KLH)as immunogen.The ELF3-KLH complex was injected into rabbits subcutaneously,and then ELF3 antibody was purified using affinity chromatography.The titer of the antibody was evaluated by ELISA.The specificity of antibody against ELF3 and immunolocalization of ELF3 were evaluated by using Western blot and immunohistochemistry.Results Rabbit polyclonal antibody against ELF3 was prepared by the immunization of ELF3-KLH complex.ELISA and Western blot results showed the antibody against ELF3 had high titer and specificity.Western blot and immunohistochemical studies demonstrated ELF3 was expressed in the mouse heart,liver,brain and kidney tissue,particular on the cell membrane.Conclusion The preparation of polyclonal antibody against ELF3 was successful due to its high titer and specificity;ELF3 was expressed in the mice heart,liver,and kidney,particular on the cell membrane.It will provide an excellent tool for further study on the ELF3 function.