10.3969/j.issn.1000-3606.2013.06.022

Agenesis of Corpus Callosum ; Chromosomes, Human, X ; Gene Deletion ; Glycerol Kinase ; genetics ; Humans ; Infant ; Male ; Syndrome

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.

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

Hepatic encephalopathy is a common metabolic neuropsychiatric syndrome in the development of end-stage liver disease. Since the concept of intestinal-liver-brain axis was proposed, the relationship between the pathogenesis of hepatic encephalopathy and the gut microbiota has been a hot research topic. In recent years, studies have confirmed that gut microbiota is involved in and affects various pathological processes of hepatic encephalopathy. This article combines the latest research progress at home and abroad to elaborate on the research status of regulating gut microbiota and thus interfering with the pathological process of hepatic encephalopathy, hoping to provide new ideas and methods for the intervention of hepatic encephalopathy based on the regulation of gut microbiota.

Objective To investigate the mechanism of the permeability change in the blood brain barrier(BBB)at early stage of infectious brain edema which was pretreated with LPS.Methods PMNs were isolated and purified.The concentration of Glu was measured in the supernatant without cells.The rat BMECs were primarily isolated and cultured.The permeability of BMECs waB evaluated by measuring the passing rate of 125 I-BSA via γ ray counter.The expression of NMDAR1 on BMECs was evaluated.Results The concentration of Glu in the LPS preconditioning PMNs subgroup at 5 rain time point was highest.In each group conditioned for 240 min,the permeability index of Bake average of the LPS preconditioning PMNs extract group was highest(P＜0.01).The expression of NMDARI increased in the LPS preconditioning PMNs extract group was more than that in other groups.Conclusions The research showed for the first time that the concentration of Glu increased after LPS preconditioning and the Glu excreted by PMNs can promote the permeability of BMECs and change the function of BBB,which possibly may be related the mechanism of NMDAR1 expression increased in BMECs.

OBJECTIVETo evaluate the application of shear wave elastography (SWE) combined with transition zone biopsy in the detection of prostate cancer (PCa).

METHODSA total of 489 patients with suspected PCa underwent transrectal ultrasonography (TRUS) and SWE-guided prostatic biopsy. We evaluated the role of SWE combined with transition zone biopsy in promoting the detection rate in comparison with the results of biopsy pathology.

RESULTSThe pathological results confirmed 221 malignant and 268 benign cases. Based on systematic biopsy, SWE combined with transition zone biopsy achieved a detection rate of 45. 19% , significantly higher than that of systematic biopsy alone (33.13%) (P < 0.05). The diagnostic sensitivity, specificity, and accuracy of SWE were significantly better than those of TRUS (P < 0.05). The mean elasticity (Emean) of SWE was remarkably higher for malignant than for benign lesions ([40.1 ± 9.5] vs [21.6 ± 8.3] kPa, P < 0.05). With 28.5 kPa as the threshold of the Emean value, the area under the ROC curve was 0. 899, and the diagnostic sensitivity and specificity were 88.71% and 86.23%, respectively.

CONCLUSIONSWE combined with transition zone biopsy could significantly improve the detection rate of prostate cancer.

Elasticity Imaging Techniques ; methods ; Humans ; Image-Guided Biopsy ; methods ; Male ; Prostate ; pathology ; Prostatic Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; ROC Curve ; Sensitivity and Specificity

Objective To investigate the molecular mechanism for change in permeability in brain microvascular endothelial cells (bEnd.3) induced by lipopolysaccharide (LPS). Methods Monolayers of bEnd.3 were exposed to LPS,in the presence or absence of exoenzyme C3 transferase. We monitored the monolayer barrier integrity by transendothelial electrical resistance assay (TEER),activity of RhoA by pull down assay,NF-κB by luciferase reporter assay,and F-actin dynamic structure by Rhodamine-phalloidin staining. Results Incubation of monolayers with LPS caused substantial barrier hyperpermeability. Under the had been treated for 3 and 12 h with LPS (P＜0.05). Such effects could be inhibited partly by pretreatment of RhoA inhibitor exoenzyme C3 transferase. LPS activated RhoA and NF-κB at 0.5 h. The C3 transferase could significantly reverse the NF-κB activation (P＜0.05). The F-actin rearrangments displayed in a time-dependent manner and occurred originally after the stimulation of LPS for 3 h,which could be diluted by the pretreatment of C3 transferase as well. Conclusion LPS induces the disruption of F-actin cytoskeleton and brain microvascular endothelial barrier integrity,in part,through RhoA and NF-κB activation. The mechanism underlying this pathophysiological effect of RhoA is to influence the disruption of the F-actin cytoskeleton by regulating NF-κB activites.

[ 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.