Objective To investigate the influence factor of the recovery time of oculomotor nerve palsy (ONP) after traumatic carotid-cavernous sinus fistula (TCCF) treated by balloon embolization.Methods The clinical data of 76 patients with ONP after TCCF were retrospectively analyzed.All patients accepted intravascular balloon embolization treatment.Nonparametric test was applied to make single factor analysis of the influence factor of ONP recovery time,and linear regression analysis was applied to make multiple factor analysis.Results Seventy-six patients (100.0％) had a perfect occlusion for orificium fistulae after operation immediately,and 73 patients (96.1％) retained the internal carotid artery.Patients were followed up for 6-70 months,with an average of 34.2 months and no death cases.Seventy patients (92.1％) succeeded for embolization at the first time,and 6 patients (7.9％) relapsed after embolization for 6 weeks.The reasons of relapse was balloon leak,and no patients recurred after twice embolization.Seventy-six patients (100.0％) had recovery from ONP,and recovery time was (42.17 ± 32.39) d.The single factor analysis showed that the courses of diseases,fistula location,eye-tubercle location,degree of ONP,balloon quantity,state of internal carotid artery were the factor affecting the ONP recovery time (P ＜ 0.01 or ＜ 0.05).The linear regression analysis showed that the courses of the disease,fistula location,degree of ONP,balloon quantity were independent factor affecting the ONP recovery time (P ＜ 0.01).Conclusions Intravascular balloon embolization in the treatment of ONP after TCCF is safe and reliable.The courses of diseases,fistula location,degree of ONP and balloon quantity are the influencing factor of the oculomotor nerve functional recovery time,and should be given enough attention.

Vascular endothelial growth factor 165 (VEGF(165))-mediated autocrine stimulation of tumor cells enhances the progression to a malignant phenotype. VEGF(165)b competes with VEGF(165) and binds to vascular endothelial growth factor receptor (VEGFR), resulting in inhibition of downstream signal transduction pathways. This study was designed to investigate the role of VEGF(165)b in the migration and invasion of human lung adenocarcinoma A549 cells. The full-length of VEGF(165)b was constructed and cloned into an expression plasmid (pVEGF(165)b), and then transfected into A549 cells. Dimethylthiazolyl- 1 -2, 5-diphenyltetrazolium bromide (MTT) assay was used to detect the effect of VEGF(165)b on proliferation of transfected cells. Reverse transcription polymerase chain reaction (RT-PCR) was employed to examine the effect of VEGF(165)b on the expression of VEGF(165) in transfected cells. Wound-healing assays were used to investigate the effect of VEGF(165)b on migration of transfected cells. Matrix metalloproteinase (MMPs) activity assay and in vitro invasion assay were used to determine the role of VEGF(165)b in invasion of transfected cells. There was no significant change in proliferation of A549 cells after transfection of pVEGF(165)b, but the expression of VEGF(165), migration and invasion in A549 cells were inhibited. Furthermore, exogenous VEGF(165)b inhibited the activity of MMP9 in the supernatant of A549 cells and the subsequent invasion capacity of those cells. We therefore conclude that exogenous VEGF(165)b can inhibit the expression of VEGF(165), as well as the migration and invasion of A549 cells, but has no effect on the proliferation of A549 cells.

Objective To explore the effect of SFI in radiation-induced mice brain injury after 20 Gy cranial radiation.Methods The mice were divided into three groups:(1) control group,(2) RT-only group:the whole brain was irradiated with a dose of 20 Gy,(3) RT and SFI group:SFI at 20 ml/kg/d from 4 weeks after 20 Gy cranial radiation theraty(CRT).Results Morris water maze test showed that the latency of the irradiated group was longer than control group and SFI improved the cognitive function of mice (t =6.34,6.70,P ＜0.05).The expression of TNF-α reached to the highest level at 3 h after irradiation,and then it decreased but got the second higher level again at 4 weeks after irradiation.The expression of IL-1 β reached to the highest level at 72 h after irradiation and decreased until 4 weeks after irradiation.SFI decreased both expressions of TNF-α (t =11.34,9.70,6.07,P ＜ 0.05) and IL-1 β (t =12.27,5.70,7.52,P ＜ 0.05).Immune florescence staining showed that SFI reduced the number of activated microglia (t =12.35,8.64,7.82,P ＜ 0.05)and inhibited the translocation of p65 of microglia after irradiation.Conclusions Findings suggest that SFI may decrease microglial activation and suppress the expression of TNF-α and IL-1β by inhibiting the translocation of NF-κB p65 and then attenuate irradiation-induced brain injury.

Objective To explore the inhibitory effects of Tanshinone Ⅱ A on the radiationinduced microglia activation and the possible mechanism.Methods Microglia cells BV-2 were irradiated with 2,4,8,16,and 32 Gy doses or sham-irradiated in presence or absence of 1.0 μg/ml Tanshinone Ⅱ A for 12 h,respectively.The effects of Tanshinone Ⅱ A on radiation-induced pro-inflammatory cytokines were evaluated using real-time PCR.The expression level of NF-κB p65 in cytoplasm and nucleus was measured by using Western blot.Immunofluorescence staining and confocal microscopy analysis were applied to detect the expression of γ-H2AX and p65 post-irradiation.Results The microglia cells were activated at 16,32 Gy post-irradiation.Radiation-induced release of the pro-inflammatory cytokines in BV-2 cells was detectable after irradiation.Tanshinone Ⅱ A decreased radiation-induced release of proinflammatory cytokines(t=5.56,P ＜ 0.05).Furthermore,western blotting showed that Tanshinone Ⅱ A could attenuate the nuclear translocation of NF-κB p65 submit post-irradiation.Immunofluorescence staining showed that γ-H2AX foci formation while p65 translocation into nucleus post-irradiation.Conclusions Tanshinone Ⅱ A exerts anti-inflammatory properties by suppressing the transcription of proinflammatory cytokine genes that might be associated with NF-κB signaling pathway.It is postulated that irradiation causes immediate cellular reaction and DSB triggers the molecular response which leads to NFκB pathway activation.

Objective:AFM1-BSA and AFM1-OVA were synthesized and then identified in this experiment.Methods: Using oximation method ,AFM1 was transformed to oxime compounds while the reaction process was monitored via TLC method aiming to identify the compounds.Coupled with carrier protein BSA and OVA respectively , we obtained AFM1-BSA and AFM1-OVA, then identified synthetic antigen via UV spectrophotometry and SDS-PAGE.Antigens were injected into experimental animals , finally obtaining the murine multi-antiserum.Eventually , the multi-antiserums were detected via indirect inhibition ELISA method to judge whether the antigens were effectively or not.Results:After oximation reaction ,the migration distance of oxime compounds in the thin layer plate was shorter.The maximum absorption peak of AFM1-BSA occurred in 274 nm,and was inconsistent with both UV absorption peaks of BSA and AFM 1.The electrophoretic velocity of AFM 1-BSA was less than that of BSA.All the titers of three immunized mice were 1×10-4 approximately;the multi-antiserum from No.3 sample had the best sensitivity ,its IC50 was 359.9 ng/ml.Conclusion:In this study,we obtained AFM1 artificial antigen and murine multi-antiserum of high sensitivity.

Objective To explore the inhibitory effects of Corilagin on the production of proinflammatory cytokines in microglia induced by radiation. Methods The cytotoxicity of Corilagin was measured by MTT assay. Microglia BV-2 cells were irradiated 0 or 32 Gy after pretreated with Corilagin for 12 hours. Realtime-PCR was used to detect the mRNA levels of inflammatory cytokines, such as IL-1β,TNF-α on several time-points. The content of nitric oxide (NO) was determined with nitrate reductase method. The translocation of NF-κB was measured by Western blot and immunocytochemical stain.Confocal microscopy was used to observe the expression of Iba-1 and Nemo. Results No cytotoxicity was detected on BV-2 cells with 1-10 μg/ml Corilagin. Iba-1 expression in microglia cells was activated by irradiation, the expression levels of inflammatory cytokines, such as IL-1β, TNF-α and NO were also elevated. Whereas, the production of IL-1 β, TNF-α in activated microglia cells was significantly inhibited with 5 μg/mL corilagin ( tIL-1β = 6. 341, tTNF-α = 3.41 1, tNO = 3. 134, P ＜ 0. 05 ). Corilagin significantly inhibited the expression of Nemo and the translocation of NF-κB p65. Conclusion Corilagin could inhibit the activation of irradiated microglia cells and down-regulate the expression of inflammatory cytokines, via inhibition of the NF-κB signaling pathway.

AIM:To investigate the cytological basis and differentiating conditions of human bone marrowmes-enchymal stem cells(hMSCs) differentiated into cells of the endothelial lineagein vitro.METHODS:hMSCs were isolatedby density gradient centrifugation and fractionated on a 1 073 g/L Percoll.The combination of VEGF165and various matrixproteins including fibronectin(FN) and typeⅠ collagen(Col) was used to induce hMSCsin vitro.Cells were character-ized by immunohistochemistry,cytochemistry,FACS and ultrastructure to identify and detect the differentiated populationand markers.RESULTS:hMSCs was positive for KDR.PAS reaction was positive and ultrastructure of hMSCs showedglycogen-pool in ectoplasm.Glycogen reducing or disappear suggested that stem cells have occurred differentiation.In-duction of hMSCs resulted in the increase of KDR,?1integrin and CD34.CONCLUSION:hMSCs were induced to atransit population(TP) that differentiated toward the endothelial progenitor cells(EPC),but not a really EPC.hMSCspedigree diagram of differentiation was hMSCs→TP→EPC→endothelial cells(ECs).

Objective To investigate the clinical significance of leukocyte count and neutrophil count change on delayed traumatic intracranial hemorrhage(DTIH).Method Collected the peripheral venous blood of l 16 eases of DTIH and 123 cases of non-DTIH between June 2005 and Deeember 2007.Compared the leukocyte count and neutrophil count in all cases by different groups.Results The leukocyte count and neutrophil count of the DTIH patients whose first CT scan were negative[(13.35±6.72)×109/Land(12.78±6.43)×109/L,respectively]were significantly higher than those of head injury whose CT scan werenegative allthetime[(9.72±3.09)×109/Land(7.64±2.93)×109/L,respectively].The difference was significant (P＜0.01).There were no statistical significance of the leukocyte count and neutrophil count between DTIH patients and non-DTIH patients whose first CT scan were positive(P＞0.05).Conclusions As an independent guideline,the leukocyte count and neutrophil count may help the forepart diagnosis of DTIH patients whose first CT scan were negative.

AIM: To investigate the cytological basis and differentiating conditions of human bone marrow mesenchymal stem cells (hMSCs) differentiated into cells of the endothelial lineage in vitro. METHODS: hMSCs were isolated by density gradient centrifugation and fractionated on a 1 073 g/L Percoll. The combination of VEGF165 and various matrix proteins including fibronectin (FN) and type I collagen (Col) was used to induce hMSCs in vitro. Cells were characterized by immunohistochemistry, cytochemistry, FACS and ultrastructure to identify and detect the differentiated population and markers. RESULTS: hMSCs was positive for KDR. PAS reaction was positive and ultrastructure of hMSCs showed glycogen- pool in ectoplasm. Glycogen reducing or disappear suggested that stem cells have occurred differentiation. Induction of hMSCs resulted in the increase of KDR, β1 integrin and CD34. CONCLUSION: hMSCs were induced to a transit population (TP( )) that differentiated toward the endothelial progenitor cells ( EPC), but not a really EPC. hMSCs pedigree diagram of differentiation was hMSCs→TP→EPC→endothelial cells (Ecs).

BACKGROUNDTo determine the antigenicity of HGV NS5 recombinant proteins expressed in E.coli.

METHODSHGV NS5a,NS5b and core/NS5b fusion genes were cloned into pThioC vector. Three expression plasmids were transformed into JM109(DE3) competent cells then expressed with induction by IPTG. Western blot and ELISA were used to determine the antigenicity after the three recombinant proteins were purified.

RESULTSAfter identification by restriction enzyme and sequencing, it was confirmed that the expressed was target proteins espected. Purified expression proteins were found strongly immunoreactive among anti HGV positive sera by Western blot and ELISA. Compared with mixed recombinant antigen (including core, NS5a synthetic peptide and NS3 recombinant proteins), in the 22 positive sera detected with mixed antigen, 68%(15/22), 90%(20/22) and 73%(16/22) were positive by P5a,P5b and Pc?5b antigens; In the 70 negative samples with mixed antigen, 7%(5/70), 1%(1/70) and 6%(4/70) were positive by P5a, P5b and Pc?5b antigens. The positive alone was found among RTPCR positive specimen using these recombinant antigens.

CONCLUSIONSNS5 gene expressed in E.coli?which couldn't be covered with other regions of antigens was one of the essential epitopes to HGV immunologic diagnosis.

Antibodies, Viral ; blood ; Antigens, Viral ; blood ; Epitopes ; immunology ; GB virus C ; genetics ; immunology ; Humans ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; immunology ; Viral Nonstructural Proteins ; genetics ; immunology