Objectives To analysis the expression difference of eIF4E in bone marrow between the acute myeloid leukemia (AML) and the non-tumor patient,before and after induction therapy,and the different subtypes of AML,and to explore the relation between eIF4E and other molecular biology abnormalities in AML.Methods The bone marrow specimens of newly diagnosed AML and non-tumor control patients were collected.The expression level of eIF4E was detected by RT-PCR.Results The positive rate of eIF4E was 65.2 ％ (101/155) in AML.eIF4E turned to negative in 12 patients (17.6 ％) who got complete remission after induction therapy.eIF4E was negatively expressed in bone marrow of the nontumor control patients.The positive rates of eIF4E were 75.0 ％ (15/20) and 80.8 ％ (21/26) in M4 and M5 type AML,respectively,and was 59.6 ％ (65/109) in other subtype AML (P ＞ 0.05).FLT3/ITD gene mutation was found in 26 cases of newly diagnosed AML.The eIF4E expression and FLT3-ITD gene mutation were independent each other (P ＞ 0.05).Conclusions eIF4E is positive in most of the newly diagnosed AML and turns to negative in part of AML achieving complete remission.The expression of eIF4E is no difference among different subtype of AMLs.eIF4E and FLT3-ITD gene mutation are independent each other.

Objective To investigate the feasibility of the functional pulmonary lobectomy (FPLT) by studying the blood-gas and morphology of the FPLT model. Methods 18 healthy dogs were divided into three groups randomly: Group A (n=6): the target bronchus and lung bubbles were filled with emulsion of iodine oil and pingyangmycin(PYM)and then target bronchus was occluded with polymethylmethacrylate(PMMA).Group B (n=6): the target bronchus was only occluded with PMMA and Group C (n=6): the target pulmonary lobe was resected. Artery blood gas were measured at the time of pre-operation and post-operation immediately and then 1st, 2nd, 3rd and 4th week after operation respectively. Chest radiolography and histology and bacterial culture of tissue of target lung lobe were made after 4 weeks. Results There was a significant difference in artery blood-gas among 3 groups pre-operation compared with post-operation immediately (P0.05) compared with 1st, 2nd, 3rd and 4th week after operation. It showed atelectasis radiologically and fibrosis of target lung lobe histologically and no bacterium grew in target tissue the 4th week after operation. There were 3 cases of lung atelectasises but no pulmonary fibrosis in group B. Conclusion FPLT may be obtained after the target bronchus and lung bubbles were filled with emulsion of iodine oil and PYM and then target bronchus were occluded.FPLT is a minimal invasive, safe and effective procedure and might partially replace the surgical pulmonary lobectomy in future.

Objective To investigate the role of vitamin D receptor (VDR) in the protection of bufalin on podocyte injury induced by adriamycin (ADR).Methods (1) In vitro:the toxic effect of different concentrations of bufalin (10-9,10-8,10-7,104 mol/L) on podocyte was evaluated by lactate dehydrogenase (LDH) test;Annexin V-FITC and RT-PCR were utilized for podocyte apoptosis and VDR mRNA level respectively.Western blotting was used to analyze the protein expression of VDR and nephrin.SiRNA intervene was also applied to evaluate the role of VDR in bufalin's protective effect on podocyte injury induced by ADR.(2) In vitro:24 SD rats were randomly divided into three groups:control group,ADR group and ADR+bufalin group.TUNEL assay was applied to detect the apoptosis of podocytes in the kidney.Immunofluorescence and transmission electron microscope (TEM) were applied to analyze the expression of VDR and the ultrastructure of the glomerulus.Results Bufalin concentration lower than 10-7 mol/L had no toxicity on normal podocyte.Bufalin reduced the urinary protein excretion (P ＜ 0.05),alleviated the removal of podocyte foot processes and attenuated the changes in nephrin expression in the glomerulus of the adriamycin (ADR) rats (P ＜ 0.05).Bufalin notably inhibited the down-regulation of VDR in protein levels on the glomerulus of the ADR rats.Additionally,bufalin inhibited the down-regulation of VDR in both mRNA levels and protein levels (P ＜ 0.05),nephrin protein expression (P＜ 0.05),and apoptosis induced by ADR in cultured podocytes.Additionally,VDR specific siRNA intervene abolished the protective effect of bufalin in ADR-induced podocyte injury.Conclusion Bufalin can alleviate ADR-induced podocyte injury via enhancing VDR expression.

AIM:To determine if lysophosphatidic acid(LPA)regulates the proliferation of astrocytes(AS)and to approach the mechanism of the process.METHODS:The cerebral AS of the neonatal SD rats were cultured in vitro and divided randomly into control group,PKC excitomotor(PMA)group,LPA group,PKC-? inhibitor(Ro31-8220)group,Ro31-8220+PMA group and Ro31-8220+LPA group.The proliferation of the cells was detected by MTT assay and flow cytometry(FCM).The concentration of intra-cellular calcium ion of the cells([Ca~(2+)]_i)which were labeled with Fura-2/AM was determined by ultraviolet spectrophotometer.The change of PKC-? inside the cells was observed by Western blotting.RESULTS:LPA and PMA stimulated the proliferation of AS,they also enhanced the expression of PKC-? and increased the concentration of [Ca~(2+)]_i.After pretreated with Ro31-8220,the abilities of LPA that mentioned above were decreased.The change of [Ca~(2+)]_i was associated with the diversity of PKC-?.CONCLUSION:LPA promotes the proliferation of AS via the way of PKC-? and Ca2+.

AIM:To understand the effects and approach the mechanisms of matrix metalloproteinase-9(MMP-9)and cystoskeleton actin on the permeability increasing of blood-brain barrier(BBB)model which was induced by hypoxia/ischemia status in vitro.METHODS:The BBB model was build by the co-culture of cell ECV304 and astrocytes in vitro,then divided randomly into control group,hypoxia/ischemia group and BB-1101 pretreatment group.The permeability of BBB was determined by [125I]-BSA.The expression and the disposition of actin were detected by direct-immunofluorescence and Western blotting.BB-1101,the MMPs inhibitor,was used to investigate if MMP-9 participate the process of the increasing of BBB models' permeability in hypoxia/ischemia status.RESULTS:Post-stimulation of hypoxia/ischemia for 5 h,the permeability of [125I]-BSA and amount expression of MMP-9 in hypoxia-ischemia group was increased compared with control group(P

BACKGROUND:The levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α) are increased during infectious brain edema, and are positively relevant to the degree of brain damage. However, whether TNF-α can enhance blood brain barrier (BBB) permeability remains unclear, especially in vitro.OBJECTIVE: To understand the changes and possible mechanism of the BBB permeability induced by TNF-α in vitro.DESIGN: Randomized controlled cell model study in vitro.SETTING:Department of Pediatrics, Xiangya Hospital, Central South University; Department of Biochemistry, Xiangya Medical College, Central South University.MATERIALS: Twenty 7-day-old healthy Sprague-Dawley rats, of clean grade and either gender, were provided by the Animal Center, Xiangya Hospital, Central South University. TNF-α was purchased from sigma Company; DMEM fluid medium and fetal bovine serum were purchased from Hyclone Company; Y-27632 was purchased from Alexis Company,and rabbit anti-human factor Ⅷ -related antigen was purchased from Zymed Company; Mouse anti-rat glial fibrillary acidic protein (GFAP) was purchased from Neomarkers. Other biochemical reagents were imported (Sigma Company).METHODS: This experiment was carried out in the Xiangya Hospital, Central South University between March 2004 and April 2005. Brain microvascular endothelial cells and astrocytes were co-cultured 10 days to set up rat models of BBB in vitro. Then, the cells were divided into 4 groups: model group(BBB models were prepared), TNF-α group ( BBB model incubated with 0.01 g/L TNF-α for 5 hours), Y-27632 pretreated group ( BBB model incubated with 30 μmol/L Y-27632 for 1 hour before 0.01g/L TNF-α challenge ) and Y-27632 control group (BBB models only incubated with Y-27632 as those in the Y-27632 pretreated group). The effect of TNF-α on BBB permeability was observed by detecting the 125 I -BSA, which passed through the inserts at each time point (30,60,120 and 240 minutes) using .γradioimmunoassay counter.MAIN OUTCOME MEASURES: The 125 I -BSA, which passed through the inserts in rat models of BBB at different time points after intervention.RESULTS: The 125 I -BSA, which passed through the inserts in rat models of BBB, was all significantly higher in the TNF-α group than in the other groups at 30, 60, 120 and 240 minutes after intervention, respectively (P ＜ 0.01), and reached the peak at 240 minutes; The 125 I -BSA, which passed through the inserts, was lower in the Y-27632 pre-treated group than in the TNF-α group at 30 and 60 minutes after intervention (P＜ 0.01). There was also significant difference in 125 I -BSA permeation between Y-27632 pretreated group and Y-27632 control group after 120 minutes (P ＜ 0.05).CONCLUSION: TNF-α can increase BBB permeability, and Y-27632 pretreatment can early reverse the effect of TNF-α on BBB permeability.

Objective:To evaluate the accuracy and influence factors of a triple-united method used for the determination of root canal working length.Methods:Root canal working length was detemined by manual measurement,tooth average length and radiographic measurement(triple-united method).In vitro experiment was conducted in 40 teeth and clinical study in 96 teeth by experienced specialists and postgraduates.Results:The accuracy of triple-united method was 82.5% and 62.5% by spcialist and poststudents in vitro, 87.1% and 75.5% in clinic,respectively. The accuracy was interfered by experience of the examiner, tooth integrity and apical disease.Conclusion:Triple-united method can meet the clinic requirement in the determination of root canal length.

Objective To understand the changes and possible mechanism of the blood brain barrier(BBB) permeability induced by tumor necrosis factor(TNF-?) in vitro.Methods BBB model was established by coculturing allogenic brain microvessel end othelial cell(BMEC) and astrocyte(AS).The BBB model was divided randomly into normal control group,TNF-? group and Y-27632 pretreatment group.The changes of BBB permeability were evaluated by Gamma radioim muno assay counter.Results The Gamma radioimmuno assay indicated that the marker,~(125)I-BSA,across the BBB model in vitro was significantly increased after TNF-? treatment compared with control group,Y-27632 pretreatmen could prevent the permeability of BBB induced by TNF-?(P

[ ABSTRACT] AIM:To study the effect of interleukin-1β( IL-1β) on neuron activation during the process of me-dial temporal lobe epilepsy ( MTLE ) .METHODS: IL-1β, rapamycin [ an inhibitor of mammalian target of rapamycin (mTOR)]and lentiviral transfection to knockdown PI3K-p85 were used to pre-treat the neurons.The protein levels of PI3K-p85, p-Akt, p-p70S6K and MAP2 were detected and the relationship among the tested cytokines was analyzed.The neuron endocytosis was observed in each group.RESULTS:IL-1βincreased the protein levels of PI3K-p85, p-Akt and p-p70S6K, up-regulated the expression of PI3K-p85 binding with IL-1RI in the neurons, and increased the neuron endocyto-sis compared with control group (P<0.05) .These processes were inhibited by rapamycin and silence of PI3K-p85 (P<0.05).Inhibition of the PI3K-p85 binding to IL-1RI decreased the protein levels of p-Akt, p-p70S6K and MAP2 which were increased by IL-1βstimulation (P<0.05).CONCLUSION: IL-1βactivates PI3K-p85 by binding with IL-1RI to promote the activation and proliferation of neuron synapses via PI3K/Akt/mTOR signaling pathway, which might be one of the mechanisms in MTLE chronic progress.

Objective This study was designed tO survey the prevalence,distribution of nonmotor symptoms(NMS)in Parkinson disease(PD)and their relationship with disease severity and duration. Methods We used Unified Parkinson Disease Rating Scale(UPDRS)part Ⅲ,V,the PD NMS Questionnaire,activities of daily living(ADL)and mini-mental state examination(MMSE)to assess motor and non-motor symptoms in 81 patients with Parkinson disease. Results In PD,a range of NMS occurred across all disease stages.Each PD patient had 7 different NMS on average.The problems of remembering thing,constipation and nocturia ranked top 3 of NMS in patients with PD.There was a significant association of total NMS score with H-Y stage(P<0.01)and UPDRS Ⅲscore(P<0.01).There was a negative correlation between NMS score and ADL score(P