Based on the current situation of teaching for laboratory animal science and taking the teaching practice of the teaching and research section of lab animal science in Liaoning Medical University as a case.,a series of teaching reforms were performed such as teaching content,teaching measures and examination manners,and the teaching reform effects were evaluated,which aimed at establishing the foundation of an consummate teaching pattern for laboratory animal science.

Objective To express functional haemegglutinin(HA)protein in two different bacularvirus expression systems.Methods The whole open reading frame of A/Sichuan/1/2009(H1N1)HA was obtained by synthesis,and the HA protein were expressed in insect cells by two different bacularvius expression systems:BaculoGold system and Bac-to-Bac system. Soluble HA protein was identified by Western blot and haemegglutination test. Results The correct full length of HA gene was obtained and cloned into pAcGP67B and pFAST Bacl vectors,respectively.After 3 rounds of virus amplifyjng by re-infection of Sf9 cells,the HA protein was detected in supematant of BaculoGold system and in intracellular of Bac-to-Bac system which is much better than the former.Purified HA protein was positive not only identified by Western blot,but also detected by haemegglutinin test. Conclusion Functional HA protein was successfully expressed in two distinct bacularvirus expression systems,of which the Bac-to-Bac bacularvirus expression system is more suitable for expression of A/Sichuan/1/2009(H1N1)HA protein.

Hemangioendothelioma ; complications ; Humans ; Infant, Newborn ; Kasabach-Merritt Syndrome ; complications ; Sarcoma, Kaposi ; complications

Objective:To investigate the differentially expressed miRNAs in serum collected post operation and compared these miR-NAs with those collected pre-surgery among patients suffering from glioblastoma multiform (GBM) and undergoing regular clinical fol-low-up. These miRNAs may be potential biomarkers for the post-operative evaluation of patients with GBM. Methods:Forty-eight pa-tients with GBM and clinical pathological diagnosis were enrolled in this study. In the initial biomarker screening stage, total RNAs were extracted and subjected to Solexa sequencing to select miRNAs with significantly altered expression pre-and post-operation. Some of these differentially expressed miRNAs were chosen and verified through TaqMan probe-based qRT-PCR assay. A t-test was performed to determine the miRNAs that satisfied the two criteria, namely, fold change>2 and P<0.05. All of the patients were fol-lowed-up, and survival data were collected. The patients were then classified into two groups, namely, long-and short-survival groups, on the basis of the median of the miR-30e expression levels in the sera collected post-operation. Kaplan-Meier method and Log-rank test (SPSS version 19.0, IBM) were employed to determine the possible relationships between miR-30e expression levels in the sera collected post-operation and patients' overall survival. Results: Solexa revealed 63 differentially expressed miRNAs. Four miRNAs, namely, miR-26b, miR-30e, miR-129-3p, and miR-206, were selected on the basis of previous and present findings. These miRNAs were then verified in the RT-qPCR phase. Among these miRNAs, only miR-30e was significantly upregulated post-operation. The serum miR-30e expression level post-operation was not significantly associated with the overall survival of the patients. A low miR-30e expression level corresponded to prolonged survival. Conclusion:miR-30e was upregulated in the sera collected post-operation from patients with GBM. This miRNA may be negatively related to the tumor load of these patients. The miR-30e expression level in the serum col-lected post-surgery serum was not significantly associated with overall survival. Therefore, miR-30e may serve as a novel potential non-invasive biomarker for the post-operative evaluation of patients with GBM.

Objective To monitor state of environment control and life support system (ECLSS), and to diagnose faults and provide decision support in manned space flight. Methods According to the principle of decision support system, neural networks and the expert system, the foundational constructional structure and work flow of the ECLSS decision support system(EDSS) were designed,fault tree analyze(FTA) method was used to sort all of the faults and to form the expert knowledge framework, expert system and neural networks were integrated to adaptive learning from experience. Results In the two-men-five-days SZ-6 manned space flight, this software system worked well, and had reduced the difficulty and intensity of ground flight control and had provided good technological support for the consummation of this flight. Conclusion For the first time, the ECLSS decision support system is designed and implemented by using database and expert system technology. It can judge parameters state and diagnose faults exactly and rapidly, and the data models are designed correctly to give the rational result of parameter prediction.

Antipsychotic Agents ; poisoning ; Clozapine ; blood ; poisoning ; Exchange Transfusion, Whole Blood ; methods ; Humans ; Infant, Newborn ; Male ; Treatment Outcome

Objective To investigate the relationship of the initiative-aggressive behavior and D2S1338,D19S433 loci.Methods PCR and electrophoresis method were used to conduct genotype analysis on D2S1338 and D19S433 in the peripheral blood of 187 male initiative-aggressive violent offenders and 459 healthy men living in Jiangsu area.Results D2S1338 and D19S433 loci in initiative-aggressive behavior group and healthy group were found to coincide with Hardy-Weinberg equilibrium (P ＞ 0.05).There were significant difference in locus D19S433 (P ＜ 0.05)between initiative-aggressive behavior group and healthy group,but not on locus D2S1338 (P＞0.05).Univariate analysis showed significant differences at allele 14.2 and genotype 14-14 on locus D19S433 between the two groups (P =0.0011,P =0.0008) with the OR values being 0.50 (95 ％ CI:0.33-0.76) and 3.49(95％ CI:1.62-7.52),respectively.Conclusion Locus D19S433 may be related to with initiative-aggressive behavior with allele 14.2 being the resistant factor and genotype 14-14 being the susceptible factor.

Niemann-Pick disease type C (NPC) is a fatal,neurovisceral lipid storage disease,neuropathologically characterized by cytoplasmic sequestration of glycolipids in neurons,progressive neuronal loss,neurofibrillary tangles (NFTs) formation,and axonal spheroids (AS).Cytoskeletal pathology including accumulation of hyperphosphorylated cytoskeletal proteins is a neuropathological hallmark of the mouse model of NPC (npc mice).With a goal of elucidating the mechanisms underlying the lesion formation,we investigated the temporal and spatial characteristics of cytoskeletal lesions and the roles of cdc2,cdk4,and cdk5 in lesion formation in young npc mice.Cytoskeletal lesions were detectable in npc mice at three weeks of age.Importantly,concomitant activation of cdc2/cyclin B 1 kinase and accumulation of a subsequently generated cohort of phospho-epitopes were detected.The activation of cdk4/cyclin D1 and cdk5/p25 kinases was observed during the fourth week of life in npc mice,and this activation contributed to the lesion formation.We concluded that the progression of cytoskeletal pathology in npc mice older than four weeks is accelerated by the cumulative effect of cdc2,cdk4,and cdk5 activation.Furthermore,cdc2/cyclin B1 may act as a key initial player one week earlier.Targeting cell cycle activation may be beneficial to slow down the NPC pathogenesis.

OBJECTIVETo discuss the protective effect of alkaloids from Piper longum (PLA) in rat dopaminergic neuron injury of 6-OHDA-induced Parkinson's disease and its possible mechanism.

METHODThe rat PD model was established by injecting 6-OHDA into the unilateral striatum with a brain solid positioner. The PD rats were divided into the PLA group (50 mg x kg(-1) x d(-1)), the madorpa group (50 mg x kg(-1) x d(-1)) and the model group, with 15 rats in each group. All of the rats were orally given drugs once a day for 6 weeks. Meanwhile, other 15 rats were randomly selected as the sham operation group, and only injected with normal saline in the unilateral striatum. The behavioral changes were observed with the apomorphine (APO)-induced rotation and rotary rod tests. The number of tyrosine hydroxylase (TH)-positive cells in rat substantia nigra and the density of TH-positive fibers in striatum were detected by tyrosine hydroxylase immunohistochemistry. The content of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), glutathione (GSH), catalase (CAT), malondialdehyde (MDA), nitric oxide (NO) and nitric oxide synthase (NOS) in rat substantia nigra and striatum were measured by the spectrophotometric method.

RESULTAfter being induced by APO, PD rats showed obvious rotation behaviors, with decreased time stay on rotary rod and significant reduction in the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. The activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity significantly decreased, whereas the activities of NOS and the content of MDA, NO significantly increased. PLA could significantly improve the behavioral abnormality of PD rats and increase the number of TH-positive cells in sustantia nigra and the density of TH-positive fibers in striatum. It could up-regulate the activities of SOD, GSH-Px, CAT, the content of GSH and the total antioxidant capacity, and decrease the content of NOS and the content of MDA, NO.

CONCLUSIONAlkaloids from P. longum shows the protective effect in substantia nigra cells of 6-OHDA-induced PD model rats. Its mechanism may be related with their antioxidant activity.

Administration, Oral ; Alkaloids ; administration & dosage ; pharmacology ; Animals ; Apomorphine ; pharmacology ; Catalase ; metabolism ; Dopamine Agonists ; pharmacology ; Dopaminergic Neurons ; drug effects ; metabolism ; pathology ; Glutathione ; metabolism ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Motor Activity ; drug effects ; Neostriatum ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Oxidopamine ; Parkinson Disease, Secondary ; chemically induced ; physiopathology ; prevention & control ; Piper ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Substantia Nigra ; drug effects ; metabolism ; Superoxide Dismutase ; metabolism ; Tyrosine 3-Monooxygenase ; metabolism

Objective To explore the effect of Fas/FasL pathway on fluoride.induced apoptosis in hurnan neumbla8toma SH-SY5Y cells.Methods The cell survival rate,percentage of apoptosis,and mRNA expression levels of Fas and FasL were measured respectively after the SH-SY5Y cells were exposed to O(control),20,40,80 mg/L sodium nuoride(NaF)for 24 hours/n vitro.Furthermore,the changes of the percentage of apoptosis and mRNA expression levels of Fas and FasL in 40 mg/L NaF-treated groups incubated with activaling or neutralizing anti-Fas antibody(CH11 or ZB4)also observed respectively.Results Compared with the control group(100.00%), the cell surval rates in 40,80 mg/L NaF-treated groups[(84.63±2.57)%,(69.04±5.63)%]were significandy lower(P<0.01).The percentage of apoptosis in 40,80 mg/L NaF.treated groups[(8.54±1.95)%.(17.94±2.71)%]were higher(P<0.05)than thal in the control group[(3.32±1.33)%],and increased with the dose of NaF.NaF could up-regulate Fas and FasL mRNA expression,and increased the Fas/β-actin [40 ms/L group (0.94±0.51),80 mg/L group(0.99±0.12)]and FasL/β-actin[40 mg/L group(0.96±0.42),80 mg/L group(0.99±0.24)] ratio,compared with the control[Eas/β-actin(0.50±0.33),FasL/β-actin(0.58±0.23)],both the difference had 8tatistical significances (P<0.05).NaF and CH I 1 had a synergisfic effect on apoptosis and mRNA expression levels of Fas and FasLL(F=32.89,18.46,.14.69,P<0.01)while NaF and ZB4 had an antagonistic effect (F=5.73,24.26,10.17,P<0.05 or<0.01).Conclusion NaF exposure can cause apoptosis in SH-Y5Y cells,and the Fas/FasL pmhway may play an important role in NaF-induced apoptosis.