Cell Adhesion Molecules ; metabolism ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; physiology ; Humans ; Integrins ; metabolism ; Membrane Proteins ; metabolism ; Neoplasms ; metabolism ; pathology ; Organ Specificity ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Saphenous Vein ; cytology ; Tumor Cells, Cultured ; Umbilical Veins ; cytology

Objective To investigate the biofilm (BF)formation rule of nontypeable Haemophilus influenzae (NTHi)in vitro, and to observe the internal structure of BF by scanning electron microscope (SEM). Methods NTHi ATCC49247 was investigated in the present study,Pseudomonas aeruginosa (PA)PAO1 was cultured as positive control,at the same time blank control group was set up.The BF of the bacteria were cultured and then collected on day 1,2,3,4,5,6,and 7.The BF formation was detected by crystal violet staining and plate counting and the structure of BF formed by ATCC49247 was observed under SEM on day 3.Results The plate colony counting of biofilm BF by ATCC49247 and PAO1 raised during first 3 d, and then declined to (0.823 6±0.007 5)×107 cfu·mL-1 and (0.942 6±0.019 9)×107cfu·mL-1 respectively on day 7. The differences between two groups were statistically significant on day 3,4,5,and 6 (P<0.05).The differences between different time points in the same bacteria group were statistically significant (P<0.05).The densities of BF formed by ATCC49247 and PAO1 raised during the first 3 d.The absorbances on 570 nm wavelength (A570 )in two groups were 2.717 4±0.017 2 and 2.885 3±0.039 0 ,respectively;and then the A570 values in two groups declined to 0.151 7±0.074 5 and 1.196 9±1.108 5,respectively on day 7;the differences between bacteria groups and blank control were statistically significant (P<0.05 );the differences between two bacteria groups were statistically significant on day 3,4,5,and 6 (P<0.05);the differences between different time points in the same bacteria group were statistically significant (P<0.05).On day 3,the obvious BF formed by ATCC49247 were observed under SEM.Conclusion BF could be formed by NTHi in vitro;crystal violet staining,plate colony counting and SEM could be taken as conventional methods to detect BF.

Objective To investigate the rat bone structural changes of lower limb following lumbar nerve dorsal roots section.Methods Forty-eight mature female Wistar rats were divided into posterior radi- cotomy(PR)and comtrol groups randomly.The bilateral femoral bone mineral density(BMD)and biome- chanics characteristics were analyzed 2,4 and 8 weeks after the radicotomy.The same operation except the radicotomy was done in the sham group.Results In PR group,2,4,and 8 weeks after the radicotomy,the BMD of femur was(0.221?0.008)g/cm~3,(0.213?0.015)g/cm~3 ,and(0.216?0.105)g/cm~3 ,respective- ly;while that was(0.223?0.005)g/cm~3,(0.218?0.014)g/cm~3 ,and(0.208?0.111)g/cm~3 in control group.No significant difference was observed between the two groups(P＞0.05).In PR group,2,4,and 8 weeks after the operation,the mean maximum load in three-point bending test of femun midshaft was(93.64?8.76)N,(89.77?11.18)N and(93.21?8.74)N,respectively,and was lower than the values of the con- trol group at the same time point(95.94?6.29)N,(91.63?9.43)N,(95.57?8.64)N,However,there was no significant difference between the two groups(P＞0.05).Accordingly,there was no significant difference in the energy absorption in femun midshaft between the two groups(P＞0.05).Conclusion The selective rhizotomies of part lumbar never dorsal roots might not cause the loss of the femur BMD and the change of bio- mechanics property significantly in short period.

OBJECTIVETo study the metastasis-associated molecules differentially expressed in highly and poorly metastatic sublines and the mechanism of metastasis in lung giant cell carcinoma.

METHODSHighly and poorly metastatic sublines (PLA801D and PLA801C)were used as metastasis model. Cell motility and invasion assay in vitro were first compared between the two sublines. Then, gelatin zymography analysis was used to determine the MMP-2 and MMP-9 activity. The protein expression level of secreted MMP-2, MMP-9, TIMP-1, TIMP-2 and intracellular expression level of p53, p16, PCNA, CD44(V6) isomeride, E-cadherin, CK18, nm23-H1 as well as the mRNA expression level of MMP-2, MMP-9, TIMP-1, TIMP-2, VEGF were compared through Western blot. Semi-quantitative RT-PCR analysis was used to determine the intracellular mRNA expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and VEGF.

RESULTSThe in vitro cell invasion potential of highly metastatic subline PLA801D was significantly higher than that of poorly metastatic subline PLA801C by about 4 folds, while the cell motility potential was similar. The secreted MMP-2 activity was notably higher in PLA801D, which was initiated by the higher expression of MMP-2 at protein and mRNA level. In addition, the expression level of p53, PCNA, CK18 protein and VEGF mRNA were significantly higher, while the expression level of p16, E-cadherin and nm23-H1 protein were significantly lower in PLA801D. Some molecules such as MMP-9, TIMP-1, TIMP-2, CD44(V6) isomeride, which had been reported to be associated with tumor metastasis, were not observed to change significantly between the two sublines.

CONCLUSIONThere are significant differences in metastatic potential and phenotypes between highly and poorly metastatic sublines of lung giant cell carcinoma. Some differentially expressed molecules might be playing roles in promoting or inhibiting metastasis of lung giant cell carcinoma, which may be useful to elucidate the mechanism of metastasis.

Carcinoma, Giant Cell ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Interleukin-8 ; genetics ; Lung Neoplasms ; metabolism ; pathology ; Matrix Metalloproteinase 2 ; genetics ; metabolism ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Neoplasm Metastasis ; RNA, Messenger ; analysis ; Tissue Inhibitor of Metalloproteinase-1 ; analysis ; Vascular Endothelial Growth Factor A ; genetics

OBJECTIVE: The aim of this study was to explore the immunity in rats transplanted with adipose-derived mesenchymal stem cells (ADSCs) and acellular nerve (ACN) for repairing sciatic nerve defects. METHODS: ADSCs were isolated from the adipose tissues of Wistar rats. Sprague-Dawley rats were used to establish a sciatic nerve defect model and then divided into four groups, according to the following methods : Group A, allogenic nerve graft; Group B, allograft with ACN; Group C, allograft ADSCs+ACN, and Group D, nerve autograft. RESULTS: At the day before transplantation and 3, 7, 14, and 28 days after transplantation, orbital venous blood of the Sprague-Dawley rats in each group was collected to detect the proportion of CD3+, CD4+, and CD8+ subsets using flow cytometry and to determine the serum concentration of interleukin-2 (IL-2), tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) using enzyme-linked immunosorbent assay (ELISA). At each postoperative time point, the proportion of CD3+, CD4+, and CD8+ subsets and the serum concentration of IL-2, TNF-alpha, and IFN-gamma in group C were all near to those in group B and group D, in which no statistically significant difference was observed. As compared with group A, the proportion of CD3+, CD4+, and CD8+ subsets and the serum concentration of IL-2, TNF-alpha, and IFN-gamma were significantly reduced in group C (p<0.05). CONCLUSION: The artificial nerve established with ADSCs and ACN has no obvious allograft rejection for repairing rat nerve defects.
Allografts ; Animals ; Autografts ; Cytokines* ; Enzyme-Linked Immunosorbent Assay ; Flow Cytometry ; Interferon-gamma ; Interleukin-2 ; Mesenchymal Stromal Cells* ; Orbit ; Rats* ; Rats, Sprague-Dawley ; Rats, Wistar ; Sciatic Nerve ; T-Lymphocyte Subsets* ; Transplants ; Tumor Necrosis Factor-alpha

Fatty Liver ; Humans ; Proteomics ; methods

Fatty Liver ; immunology ; Immune Tolerance ; Liver ; immunology

OBJECTIVETo analyze the radiological parameters of the conservatively in treating distal radius fracture and investigate whether the final re-displacement can be predicted after primary reduction.

METHODSFrom January 2013 to June 2014,212 patients with distal radial fracture conservatively were treated in our hospital, 107 patients of them were excluded because of their incomplete radiological parameters;the remaining 105 patients were available for radiological were assessed after injury, there were 56 male patients and 49 female patients in this study, the average age of the patients was 51 years old (ranged from 22 to 80 years). According to AO classification, there were 47 cases of type A2 and C1, and 58 cases of type A3, C2, C3. All patients were treated by closed reduction and below-elbow cast immobilization for 4 to 6 weeks. All patients were followed up for 3 to 6 months (means 4.5 months) by X-ray, all fractures were healed. Standard AP and lateral radiographic examination was conducted before reduction and after reduction and bony consolidation,the dorsal angulation and the radial angle were measured at each time point. The linear regression was used for the analysis to find out whether the final re-displacement can be predicted after primary reduction.

RESULTSAmong 105 patients,the significant correlations were found for the dorsal angulation between the reduction time and the end time (r = 0.82) and for the radial angulation between the reduction time and end time (r = 0.85).

CONCLUSIONThe dorsal angulation and the radial angulation after complete healing can be predicted from linear the regression functions. Due to the possibility of predicting the end result, whether the fracture should receive further conservative treatment or surgical treatment can be decided immediately.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Male ; Middle Aged ; Predictive Value of Tests ; Radiography ; Radius Fractures ; diagnostic imaging ; therapy

OBJECTIVETo study the expression and localization of early growth response gene-1 (Egr-1) in macrophages after stimulation by silicon dioxide in vivo and in vitro and to discuss the role of Egr-1 in the development of silicosis.

METHODSThe expression of Egr-1 in animal model of silicosis was analyzed by using immunohistochemistry. Western-blot, immunofluorescence and RT-PCR analysis were used to detect the expression and localization of Egr-1 protein and the dynamic changes of Egr-1 mRNA in cultured macrophages RAW264.7, after stimulation by silicon dioxide.

RESULTSIn animal model with induced silicosis, there was an increased expression of Egr-1 in pulmonary macrophages. The expression levels peaked at the 14th day. In vitro, the transcription of Egr-1 increased in RAW264.7 macrophages during 15 to 240 minutes after the administration of silicon dioxide. The response peaked at 15 minutes and diminished to a minimal level at 480 minutes. Nuclear translocation was most apparent at 60 minutes, lasted till 120 minutes and diminished gradually. During the period from 60 to 120 minutes, the expression of Egr-1 protein also reached a peak.

CONCLUSIONSSilicon dioxide can activate the nuclear transcription factor Egr-1 in vivo and in vitro in macrophages. Egr-1 may thus play an important pathogenetic role in the development of silicosis.

Animals ; Blotting, Western ; Cell Line ; DNA-Binding Proteins ; genetics ; metabolism ; Early Growth Response Protein 1 ; Gene Expression Regulation ; drug effects ; Immediate-Early Proteins ; Immunohistochemistry ; Lung ; drug effects ; metabolism ; pathology ; Macrophages ; drug effects ; metabolism ; Macrophages, Alveolar ; drug effects ; metabolism ; pathology ; Male ; RNA, Messenger ; drug effects ; genetics ; metabolism ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Silicon Dioxide ; pharmacology ; Transcription Factors ; genetics ; metabolism

OBJECTIVETo discuss the role of early growth response factor (Egr)-1 and it's upstream signaling pathway in the development of silicosis.

METHODSThe expression and localization of Egr-1 were analyzed by immunofluorescence and in-situ hybridization. The activity of Egr-1 was observed in treated cells by using a reporter plasmid and EMSA, the activity of ERK1/2 in RAW264.7 incubated with SiO(2) by using a kinase assay, and further by using a kinase inhibitor assay to investigate the role of upstream kinase in the signal pathway of the activation of Egr-1.

RESULTSThe obvious increase of expression and transcription of Egr-1 was observed shortly after being treated by silica and its activity increased abruptly. There was an increase of the activity of ERK1/2 in RAW264.7 cells treated, which reached a peak at 30 minutes. The expression and transcription of Egr-1 decreased maniferstly after using kinase inhibitors.

CONCLUSIONEgr-1 expression can be induced by silica dioxide in RAW264.7 cells, and the ERK1/2, p38 kinases may take part in this process which suggest the pathway of SiO(2), ERK1/2, p38 and Egr-1 may play an important role in the development of silicosis.

Animals ; Butadienes ; pharmacology ; Cells, Cultured ; Early Growth Response Protein 1 ; biosynthesis ; genetics ; physiology ; Enzyme Inhibitors ; pharmacology ; Gene Expression Regulation ; Macrophages ; metabolism ; Mice ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Nitriles ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Signal Transduction ; Silicon Dioxide ; pharmacology ; p38 Mitogen-Activated Protein Kinases ; metabolism