ABSTRACT:Objective To study the BDNF/TrkB signaling pathway in the epilepsy model of hippocampal neurons and the regulatory effect of on it.Methods The primary hippocampal neurons cultured in vitro for 7 days were randomly divided into seven groups:control group,epilepsy group,control+BDNF group,epilepsy+BDNF group,control + miR-185 ? group,epilepsy + miR-185 ? group,and epilepsy + miR-185 ? + BDNF group.We constructed miR-185 ? lentivirus vector and observed the changes of BDNF/TrkB pathway expression after transfaction of miR-185 ? by immunohistochemistry,patch clamp technique and Western blot technique.Results Compared with the control+BDNF group,the phosphorylated TrkB (pTrkB)/TrkB value was significantly lower in epilepsy+BDNF group (P < 0.05 )and control group (P < 0.001 ).Compared with the epilepsy group,the phosphorylated TrkB (pTrkB)/TrkB value was significantly higher in epilepsy+BDNF group (P <0.05).Compared with the epilepsy+miR-185 ? +BDNF group,the phosphorylated TrkB (pTrkB)/TrkB value was significantly lower in epilepsy + BDNF group and epilepsy + miR-185 ? group (P < 0.001 ).BDNF could promote the signaling conduction and miR-185 ? could remove the inhibition of BDNF/TrkB signaling.Conclusion BDNF can activate the BDNF/TrkB signaling pathway and transfection with miR-185 ? can relieve the inhibition of BDNF/TrkB signaling pathway of epileptic state by up-regulating the expression of TrkB.

Objective To explore the construction and application of cloud-based healthcare information system (CHIS).Methods The process of CHIS employment and implementation for primary-level medical and healthcare institutions was elaborated by the example of a city after the analysis and comparison of mode difference between traditional and cloud construction.Results CHIS integrated regional medical resources and realized integrated application of medical and public healthcare resources,mobile integration of regional operation and continuous innovation in construction mode,and facilitated uniformed management,high-efficiency running and maintenance as well as business development.Conclusion CHIS contributes to enhancing medical inforrnatization in primary institutions,promoting continuous medical innovation and innovating medical informatization mode.

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.

Skull bore device has an important role in neurosurgery.This review is a summary of the development,current research status and advances of skull bore.The electrical operation power device is the emphasis in the review.It is important to develop newly operation power devices for human health.

AIM: To study the differentiation of rat bone mesenchymal stem cells (MSCs) into cardiomyocytes in vitro. METHODS: MSCs were isolated and purified from the bone marrow of rats by density gradient centrifugation and adhering to the plastic culture. The third passage MSCs were treated by 5-azacytidine (5-aza). The induced cells were evaluated by immunocytochemistry staining and RT-PCR analysis. RESULTS: After being induced by 5-aza, some MSCs became bigger and longer. The connection of the cells were formed on day 14.The direction of the cells arraying was similar gradually. The induced cells were stained positively for desmin, ?-actin and troponin I. RT-PCR showed that these cells expressed ? myosin heavy chain. CONCLUSION: 5-aza can induce MSCs to differentiate into cardiomyocytes in vitro.

AIM: To clone the full-length of 75A EST. METHODS: After the extraction of total RNA from primary cultured rat cerebellar granule cell of 7DIV in the medium containing 25 mmol/L KCl, T_4 DNA ligase-mediated 5' RACE was used to retrieve 5' unknown sequence of 75A EST, and the first round 5' RACE PCR product was subcloned into pGEM-T easy vector for sequence and homogeneous analysis. RESULTS: The first round of 5' RACE produce a 2.5 kb band, and 75A EST was identified to be partial sequence of Neuron-derived orphan receptor (Nor1) gene. After two more rounds RACE, we firstly cloned the full-length of Nor-1 cDNA. CONCLUSION: T_4 DNA ligase mediated 5' RACE is an efficient method to retrieve information about the 5' termini of mRNAs, and lay a foundation for further study which role Nor1 play in the cerebellar granule cell differentiation or survive.

Objective A surface feedback-stimulation gastric-pacing device was designed to extract gastric surface information,determine the status of gastric and surface stimulation.Methods The characteristic parameters of gastric electrical formed by processing signal and extracting parameter.By that,the state of stomach could be confirmed,and stimulus signal of stomach pace-making in different frequency and intensity could be decided as needed.During the processes of the stimulation,the variations of gastric electrical parameters can be analyzed in real-time and the stimulation parameters could be modified to achieve the self-feedback mode.Results Self-feedback model in different gastric motility analog environment could be realized.Conclusion The preliminary validation of experimental results proved the effectiveness of self-feedback of gastric pacing devices and its application prospects in some areas.

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.

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.