Objective To investigate the kinetics and phenotype of spleen dendritic cells (DC) in lipopolysaccharide (LPS)-induced acute lung injury (ALI) mice.Methods Thirty-six C57BL/6 mice were randomly (random number) divided into two groups:control group and ALI group.Spleens were harvested at the following intervals of 6,12,and 24 h after LPS or PBS administration.Lung wet weight / body weight ratio (LW/BW) was recorded to assess lung injury.Meanwhile,pathological changes were examined under optical microscope.The IL-6 level in the lung was measured by using ELISA (enzymelinked immuno sorbent assay).The DC in the spleen was measured by flow cytometry (FCM).Results (1) LPS-ALI resulted in a significant increase in LW/BW ratio.(2) Histologically,extensive alveolar wall thickening resulted from edema,severe hemorrhage in the interstitium and alveolus,and marked and diffuse interstitial infiltration with inflammatory cells were observed in the ALI group.(3) Meanwhile,the levels of IL-6 in lung tissue were significantly increased in the LPS-induced ALI mice.(4) LPS-induced ALI led to divergent kinetics of spleen DC in ALI mice.In ALI mice,spleen DC only showed a transient increase at 12 h.(5) All DC within spleens had a modest maturation in ALI mice.Conclusions LPS-induced ALI provokes a transient increase as well as modest maturation of spleen DC.

Objective To investigate the long time prognosis of liver resection or transcatheter arterial chemoembolization(TACE) of Barcelona clinic liver cancer stage C(BCLC-C) patients who have portal vein tumor thrombsis.Methods Totally 86 BCLC-C patients who satisfied our including criteria from our surgical database of People's Hospital of Yichang City from January 2000 to September 2015 were selected as the research object.According to different treatment,86 patients were divided into liver resection group(n=50) and TACE group(n=36).The general information of two groups were compared.Cox multi-factors analysis and overall survival rate were calculated.ResultsThe long-term prognosis of liver resection group was better than that of TACE group(5-year OS:26% vs.0,P<0.001).Multi-factors analysis revealed that multiple tumors (HR:1.510,95%CI[1.120,2.316],P=0.020),AST>40 IU/L(HR:0.615,95%CI[0.488,1.206],P=0.013) as well as HBV-DNA>1 000(HR:1.204,95%CI:[0.920,2.540],P=0.038)were adverse factors for prognosis.ConclusionLiver resection is better than TACE for BCLC-C patients with portal vein tumor thrombosis.However,randomized controlled trial still need to be used to further confirm our conclusion.

Objective To study the methods of three dimensional reconstruction of digitized virtual breast invasive ductal carcionma and sterical characteristic of tumor growth.Methods The specimens were sliced into 0.2 mm continuous sections on transversal plane with the computerized miller,and photographs were taken with digital camera.3DDOCTOR software was used to make three dimensional block diagram and the thin sectional and three dimentional characteristics of breast invasive ductal carcionma were observed.Results Every structure of breast in thin section was clear.Stereo image of breast solid could be shown satisfactorily.Every shape of stereo image and the structure of breast could be shown by revolving the three dimensional image in different direction,beneficial to us in understanding the dimensional growth pattern of tumor.Conclusion The sliced image of breast invasive ductal carcionma database was exact.The image of three dimentional reconstruction was clear,which could reflect the structural feature and growth pattern of breast carcinoma in the breast exactly and be conducive to the evaluation of tumor margin in breast conservation therapy.

AIM:To evaluate the repeatability of axial length (AL) and anterior chamber depth (ACD) obtained by A scan ultrasonography, and to compare AL and ACD obtained by A scan with those obtained by IOL Master.METHODS:Two hundred and fifty-seven cataract eyes of 170 patients were included.IOL Master and A scan were performed for each eye.Five measurements of IOL Master and 3 measurements of A scan were obtained.All the tested eyes were divided into 5 groups according to AL obtained by A scan:Group A (2129mm, 21 eyes).Cronbach's Alpha coefficient and intraclass correlation coefficient (ICC) were applied to evaluate the repeatability of AL and ACD obtained by A scan.Paired t test and Pearson correlation coefficient were used to analyze the differences and correlations of AL and ACD obtained by the 2 devices, respectively.Bland-Altman plots were presented to analyze the agreements of AL and ACD obtained by the 2 devices.RESULTS:All the Cronbach's Alpha and ICCs of AL and ACD values were more than 0.98.The differences of AL values between A scan and IOL Master were-0.11±0.08mm in Group A,-0.15±0.10mm in Group B,-0.19±0.15 mm in Group C,-0.29±0.16mm in Group D and-0.45±0.29mm in Group E, respectively (all P<0.01).The differences of ACD values between A scan and IOL Master were-0.10±0.16mm in Group A,-0.06±0.13mm in Group B,-0.06±0.13mm in Group C,-0.19±0.10mm in Group D,-0.18±0.21mm in Group E, respectively (all P<0.01).In all groups, the AL and ACD values obtained by A scan and IOL Master presented good correlations (all r>0.89, all P<0.01).CONCLUSION:The AL and ACD values in cataract eyes obtained by A scan were repeatable.The AL and ACD values obtained by A scan were smaller than those obtained by IOL Master.With the increase of AL values, the differences of AL values between A scan and IOL Master increased.

Objective There is little research focusing on the expression and function of bradykinin 1 receptor ( B1R ) and bradykinin 2 receptor ( B2R) after cerebral ischemia/reperfusion on the basis of diabetes .The aim of this study was to compare the ex-pression difference and function change of B 1R and B2R in non-dia-betic and diabetic rats . Methods The cerebral ischemia/reperfu-sion model was established on 41 non-diabetic and type 2 diabetic rats, the weight and the biochemical index were measured on these two types of rats .8 non-diabetic rats and 8 diabetic rats were respec-
tively assigned to two groups according to random number tables:control group and I/R 24 h group, 4 in each group.Real-time PCR was performed to observe the expressions of two receptors at 24 h after reperfusion .Then, 33 non-diabetic rats and 33 diabetic rats were randomly divided into 4 groups respectively, including sham group (n=6), saline group (n=9), B1R antagonist group (n=9) and B2R antagonist group (n=9).At 24 hours after cerebral I/R, neurological deficiency was evaluated by neurological severity scores ( NSS);infarct volume was observed by TTC staining;cell apoptosis was determined by TUNEL staining;neuron degeneration was de-tected by Fluoro-Jade C staining. Results Glucoses of diabetics at 3, 7, 14 d after model establishment [(23.45 ±5.01), (23.71 ±4.87), (22.72 ±4.11) mmol/L] were obviously elevated compared with non-diabetics [(5.77 ±0.75), (6.05 ±0.69), (7.15 ±1.09) mmol/L];blood cholesterin [(4.59 ±3.43) mmol/L] and insulin [(67.26 ±12.02) pmol/L] at 14 d after model establishment were evidently incresaed in comparison to those in non-diabetics [(1.58 ±0.37) mmol/L, (25.34 ±4.88) pmol/L] (P<0.05), while no significant difference was found in the blood triglyceride of diabetics between them (P>0.05).Compared with non-diabetics, diabetics suffered from more apparent up-regulation of B1R mRNA (P<0.01) but relatively less B2R mRNA (P<0.05) at 24 h after I/R.NSS score, infarction volume, damaged and apoptotic cells in B2R antagonis-treated non-diabetic rats at 24 h after I/R conspicuously decreased compared with saline-treated non-daibetic rats.Those indicators in B1R antagonis-treated diabeics were strikingly lessened compared with saline-treated daibetics . Conclusion I/R induced distinct up-regulation of B2R mRNA in non-diabetics and inhibiton of B 2R effectively ameliorated the infarct volume and cell injury after I/R in non-diabetics; I/R induced more notable up-regulation of B1R mRNA in diabetics and B1R antagonist exerted neuroprotective effects instead of B 2R antagonist af-ter I/R in diabetics.

[ Abstract] Objective To investigate the effect of IL-8 on the viability and migration of HepG2 cells and the potential effect of integrins (αsubunits ) on the migration of HepG2 cells.Methods HepG2 cells were stimulated by IL-8 at different concentrations ranging from 0 to 125 ng/ml in vitro.MTT assay was preformed to detect the viability of HepG2 cells.Scratch wound migration assay was used to explore the effect of IL-8 on the migration of HepG2 cells at the time points of 0, 4, 8, 12 and 24 h,respectively.Transwell assay was conducted to detect the vertical migration of HepG2 cells after stimulation with IL-8 for 24 h.The F-actin of HepG2 cells was observed by immunofluorescence analysis after treatment with IL-8 at different concentrations.The expression of αsubunits of integrins in HepG2 cells treated with IL-8 for 8h was detected using Western boltting assays.Results Compared with the control group, HepG2 cells treated with IL-8 showed improved proliferation activities, but there was no significant difference between the groups of HepG2 cells treated with IL-8 at different concentrations.The cell scratch healing assays and Transwell analysis both indicated that IL-8 promoted the migration of HepG2 cells in a dose-dependent manner.Meanwhile, the cytoskeleton of HepG2 cells was rearranged and the number of filopodium-like protrusions (FLPs) was increased rapidly after treatment with IL-8.Western blotting analysis showed that IL-8 up-regulated the expression of αsubunit of integrins and there was a concentration difference betweenαsubunits.Conclusion IL-8 promotes the migration of HepG2 cells by up-regulating the expression ofαsubunits, and different αsubunits may play different roles.

Objective To investigate the effect of progressive neurological deterioration ( PND) of cerebral watershed infarction on early prognosis. Methods The consecutive patients with cerebral watershed infarction admitted in the Department of Neurology,Jinling Hospital,Nanjing University School of Medicine and their cerebral watershed infarctions confirmed by the imaging examination from March 2009 to March 2014 were enrolled. The clinical features, laboratory indicators and imaging features of internal watershed infarction,cortical-type watershed infarction,and mixed watershed infarction were identified and analyzed. The National Institutes of Health Stroke Scale was used to score neurological deficit. The modified Rankin scale ( mRS) was used to score the prognosis of patients. Single factor analysis was used to compare the differences between the groups. At the same time,the correlation between PND and poor prognosis of cerebral watershed infarction at day 90 was analyzed by multivariable Logistic regression analysis. Results A total of 89 patients with cerebral watershed infarction were enrolled,including 43 cortical-type watershed infarctions,36 internal watershed infarctions, and 10 mixed watershed infarctions. Single factor analysis indicated that the incidences of PND of internal watershed infarction and mixed watershed infarction were significantly higher than the cortical-type watershed infarction (36. 1% [n=13],50. 0% [n=5], and 16. 3% [n=7],respectively;P=0. 018). At day 90,28 patients had poor prognosis,and mRS was (3.4±1. 0) scores at day 90. There was significant difference in the types of infarction between the patients with poor prognosis and patients with good prognosis (P<0. 05). In patients with poor prognosis, most of them were internal watershed infarctions,accounting for 50. 0% (14/28),while in patients with good prognosis,most of them were cortical-type watershed infarctions(57. 4% [35/61]). The incidence of PND in patients with poor prognosis was significantly higher than that in patients with good prognosis (57.1% [16/28] vs. 14. 8% [9/61];P<0. 05). The result of multivariate Logistic regression analysis showed that after adjustment for confounding factor, PND was independently associated with the poor prognosis of cerebral watershed infarction at day 90 (OR 6. 969,95%CI 2. 451-19. 869;P<0. 01). Conclusion Compared with the cortical-type watershed infarction, the patients with internal watershed infarction is more prone to have PND, and PND is independently correlate with the poor prognosis at day 90.

Objective To clarify the role of FLT3 signaling-dependent pulmonary conventional dendritic cells (cDCs) in the pathogenesis of lipopolysaccharide (LPS)-induced acute lung injury (ALI),and as well as the modulation effects of cDCs in vivo on the inflammatory responses to acute lung injury.Methods Thirty C57BL/6 male mice were divided into normal control group,LPS group,FLT3L pretreatment group,lestaurtinib,(a high efficient and specific blocker in FLT3 signal pathway) pretreatment group and vehicle (DMSD) control group.FLT3L and lestaurtinib were administrated subcutaneously for 5 days.Murine model of ALI was subsequently established by intra-tracheal application of LPS and lung specimens were harvested 6 h or 24 h later.The accumulation and maturation of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate lung inflammation.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Lung myeloperoxidase (MPO) activity was measured to evaluate neutrophil infiltration.Transcription factors Tbet/GATA-3 mRNA ratio was determined to estimate balance of Th1/Th2 response.IFN-γ and IL-4 were quantified to evaluate Th1-specific and Th2-specific cytokine production respectively.Results The accumulation and maturation of pulmonary cDCs peaked at 6h after LPS challenge.FLT3L pretreatment significantly stimulated the accumulation and maturation of pulmonary cDCs (P ＜ 0.05),leading to markedly deterioration of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/GATA-3 mRNA ratio were elevated (P ＜ 0.05).Furthermore,the production of IL-6,TNF-α and IFN-γwas markedly increased by FLT3L pretreatment (P ＜ 0.05).In contrast,lestaurtinib pretreatment markedly inhibited the accumulation and maturation of pulmonary cDCs (P ＜ 0.05),leading to significant improvement of LWW/BW and lung histopathological changes.Meanwhile lung MPO activity and T-bet/ GATA-3 mRNA ratio were decreased (P ＜ 0.05).Furthermore lestaurtinib efficiently suppressed the production of IL-6,TNF-α and IFN-γ (P ＜ 0.05).Conclusion This study thus demonstrated that FLT3 signaling-dependent pulmonary cDCs could control the initiation of acute lung inflammation response to LPS-induced ALI through the regulation of neutrophil infiltration and balance of Thl/Th2 response.

Objective To investigate the accumulation and maturation status of pulmonary conventional dendritic cells (cDCs) in the early phase of acute lung injury (ALI),and to explore the way of the inflammatory responses and lung injury modulated by cDCs in vivo.MethodsMale C57BL/6 mice were randomly ( random number) divided into the normal control group,6 h-ALI group and 24 h-ALI group.Murine model of ALI was made by intra-tracheal administration of lipopolysaccharide (LPS) and lung specimens were taken 6 h or 24 h later.The accumulation and maturation status of pulmonary cDCs were assessed by flow cytometry.IL-6 and TNF-α were quantified to evaluate the lung inflammation.Transcription factors T-bet/GATA-3 mRNA ratio was determined to estimate the balance between Th1/Th2 responses.IFN-γand IL-4 were quantified to evaluate Thl-specific and Th2-specific cytokine production respectively.Lung injury was estimated by lung wet weight/body weight ratio (LWW/BW) and histopathological assessment.Comparison between groups was performed using one -way ANOVA.ResultsCompared with normal control group,LPS challenge resulted in higher level of IL-6 and TNF-α,increased LWW/BW ratio and significant histopathological changes (P ＜0.01 ).The accumulation and maturation of pulmonary cDCs in 6 h-ALI group were significantly increased after LPS challenge (P ＜0.01 ),while the accumulation and maturation of pulmonary cDCs in 24 h-ALI group were significantly lower than that in 6 h-ALI group ( P ＜0.01 ).Compared with normal control group,the expression of T-bet mRNA in 24 h-ALI group was markedly enhanced ( P ＜ 0.01 ) and the production of IFN-γ was increased as well ( P ＜ 0.01 ).ConclusionsThe accumulation and maturation of pulmonary cDCs peaked within 24 h after LPS challenge,pulmonary cDCs may initiate and amplify acute lung inflammation of ALI by enhancing the Th1 immune response and ensuing cytokine production.

Objective To observe the expression of I-Ab/I-E on circulating,lung and splenic dendritic cells (DC) in acute lung injury (ALI) mice.Methods Twenty-four C57BL/6 mice were randomly divided into four groups:control group,ALI 6 h,ALI 12 h and ALI 24 h group.Blood,lungs and spleens were harvested after lipopolysaccharide or phosphate butter solution administration.The expression of I-Ab/I-E on DC was assessed by flow cytometry (FCM).IL-6 level in the lung was measured by enzymelinked immunosorbent assay (ELISA).Lung wet weight/body weight (LW/BW) was recorded to assess lung injury.Meanwhile,pathological changes were examined under optical microscope.Results (1) lipopolysac charide-induced ALI mice resulted in a significant increase in lung LW/BW ratio.(2) Histologically,widespread alveolar wall thickening caused by edema,marked and diffuse interstitial infiltration with inflammatory cells,and severe hemorrhage in the interstitium and alveolus were observed in the ALI groups.(3) The level of IL-6 in lung tissue was significantly enhanced in ALI mice.(4) FCM analysis showed that I-Ab/I-E expressions on lung DC [(73 ±9)％],and splenic DC [(81 ±8)％] were significantly higher than that on circulating DC [(24 ± 7) ％ ; P ＜ 0.05] in control mice.(5) In ALI mice,the expressions of I-Ab/I-E on peripheral blood DC were (34 ± 17)％ at 6 h,(51 ± 16)％ at 12 h,(50 ± 17)％ at24 h respectively; I-Ab/I-E expressions on lung DC were (82 ± 14)％ at 6 h,(88 ±6)％ at 12 h,(90 ±10)％ at 24 h respectively; the expressions of I-Ab/I-E on splenic DC were (88 ± 8)％ at 6 h,(89 ± 4)％ at 12 h,(93 ± 9)％ at 24 h respectively,which were also significantly higher than those on the peripheral blood DC (P ＜ 0.05).(6) The I-Ab/I-E expressions on circulating DC in ALl mice at 12 h and 24 h was significantly higher than that on circulating DC in control mice (P ＜ 0.05).(7) The I-Ab/I-Eexpressions on lung DC and splenic DC in ALI mice at 24 h were significantly higher than those on lung DC and splenic DC in control mice (P ＜ 0.05).(8) There was a significant correlation of I-Ab/I-E expression on respiratory DC with the IL-6 level and lung injury score in LPS-induced ALI group (P ＜ 0.05).Conclusions There is a dynamic characteristic in the expression I-Ab/I-E on circulating,lung and splenic DC populations in ALI mice.I-Ab/I-E on pulmonary DC seems to play an important role in the pathogenesis of ALI.