Objective: To study the cytocompatibility of nanophase alumina ceramics with osteoblasts. Methods: Alumina ceramics were prepared via wet chemistry techniques. The grain size of alumina of interest in the present study was determined by scanning electron microscopy and atomic force microscopy with image analysis software. Primary osteoblast culture was established from rat calvaria. Protein content, synthesis of alkaline phosphatase(ALP) and deposition of calcium-containing mineral by osteoblasts cultured on nanophase alumina ceramics and conventional alumina ceramics for 7, 14, 21 and 28 d were respectively examined. Results: The average surface grain size of the nanophase and conventional alumina compact formulations was 60 nm and 1.80 ?m respectively.Synthesis of ALP and deposition of calcium-containing mineral were significantly greater by osteoblasts cultured on nanophase than those on conventional ceramics after 21- and 28- day culture. Conclusions: Nanophase alumina may stimulate ALP synthesis and calcium deposition of osteoblasts.

A quartz tube reactor was designed to combine with a commercial mass spectrometer for on-line detecting the gases evolved during pyrolysis of coal and other samples with high volatile. The reliability and repeatability of this atmosphere pressure-temperature programmed pyrolysis-mass spectrum (AP-TPP-MS) system were tested by model compound and real coal sample. The results show that pyrolysis of model compound can give good response, less overlap or tailing gases evolution curves;and multi-peaks can be observed from the same gas curve of real sample. The reliability and repeatability of this system are perfect and the system can be applied to study the coals decarboxylation by comparing the CO_2 evolution curves.

AIM: To elucidate the pattern of 5-fluorocytosine(5-FC) induced apoptosis and its role in gene therapy for human pancreatic cancer. METHODS: The human pancreatic cancer SW1990 cells(CEA-producing) were infected with recombinant adenoviruses(Adex1CEA-prCD or Adex1CEA-prZ).Cytosine deaminase(CD) expression was examind by western blot. Apoptosis induced by 5-FC in human pancreatic cancer SW1990 cells genetically modified to express cytosine deaminase was investigated by applying electron microscopy, DNA electrophoresis and flow cytometry analysis techniques. RESULTS: The SW1990 cells infected with Adex1 CEA-prCD were treated with 5-FC at 100 ?mol?L -1 for 48 h, cell apoptosis occurred. Typical apoptosis morphological feature appeared and DNA ladder could be demonstrated on DNA electrophoresis. Apoptosis peak was also showed by flow cytometry. Apoptotic cells accounted for 34.6% of the cell population. Cells in G 1, S and G 2/M phase of cell cycle were 64%, 11% and 7%, respectively. CONCLUSION: The apoptosis induced by 5-FC may be a primary mechanism in CD gene therapy for pancreatic cancer.

OBJECTIVETo evaluate the therapeutic effects of VSD combined with irrigation of oxygen loaded fluid on the growth of granulation tissue and macrophage polarization in chronic venous leg ulcers.

METHODSThiry-four patients with chronic venous leg ulcers hospitalized in our department from December 2010 to July 2014 were divided into VSD group ( A, n = 11) , VSD + irrigation group ( B, n = 11) , and VSD + oxygen loaded fluid irrigation group ( C, n = 12) according to the random number table. After admissian, debridement was performed, and granulation tissue in the center of the wound was harvested during the operation. After dehridement, the patients in group A were treated with VSD only (negative pressure from -30 to -25 kPa, the same below) ; the patients in group B were treated with VSD combining irrigation of normal saline; the patients in group C were treated with VSD combining normal saline loaded with oxygen irrigation (flow of 1 L/min) . On post treatment day (PTD) 7, the VSD devices were removed. Cross observation was conducted before debridement and on PTD 7. On PTD 7, the granulation tissue in the center of the wound was harvested for histopathological observation with HE staining and Masson staining, following calculation of granulation tissue coverage rate. After debridement but before the negative pressure therapy (hereinafter referred to as before treatment) and on PTD 7, partial pressure of oxygen of the skin around the wound was measured by transcutaneous tissue oxygen tension survey meter. On PTD 7, expression of vascular endothelial growth factor (VECF) was determined with immunohistochemistry. Before treatment and on PTD 7, cells with double positive expressions of induced nitric oxide synthase plus CD68 ( type I macro- phage) and arginase 1 plus CD68 ( type II macrophage) were observed with immunofluorescence staining and quantified. Data were processed with Fisher's exact test, one-way analysis of variance, covariance analysis, paired test, and LSD test.

RESULTS(1) The gross observation showed that before debridement there was a certain amount of necrotic tissue and little granulation tissue in the wounds of patients in all the 3 groups. On PTD 7, new granulation tissue was found in the wounds of patients in all the 3 groups, and in group C its amount was the largest. (2) On PTD 7, the granulation tissue coverage rate of wounds in pa- tients of group C was higher than that of group A or B ( P <0.05 or P <0.01). (3) On PTD 7, HE staining showed that there appeared more abundant new born microvessels and fibroblasts in the wounds of patients in group C than those in groups A and B; Masson staining showed that there was more abundant fresh collagen distributed orderly in the wounds of patients in group C compared with group A or B. (4) On PTD 7, it was found that partial pressure of oxygen of the skin around the wounds in patients of group C [(40.7 +/- 4.1) mmHg, 1 mmHg = 0.133 kPa] was higher than that of group A [ (35.0 +/- 3.1) mmHg] or B [(35.4 +/- 2.7) mmHg, with P values below 0.01]; the partial pressure of oxygen of the skin around the wounds of patients in all the 3 groups was increased significantly compared with that before treatment (with values from 10.38 to 22.52, P values below 0.01). (5) On PTD 7, the expression of VECF in the wounds of patients in group C was higher than that in group A or B ( P <0.05 or P < 0.01). (6) On PTD 7, the number of type I macrophages in granulation tissue of patients was respectively 14.3 +/- 2.3, 11.5 +/- 3.0, and 10.7 +/- 2.3 per 400 times vision field in groups A , B, and C ( F = 25.14, P < 0.01), while the number in group C was less than that in group A or B ( P < 0.05 or P < 0.01). Compared with that before treatment, the number of type I macrophages was significantly decreased on PTD 7 in all the 3 groups (with values from 14.76 to 23. 73, P values below 0. 01). On PTD 7, the number of type II macrophages in granulation tissue of patients was respectively 32.7 +/- 3.2, 35.1 +/- 3.3 , and 41.3 +/- 3.2 per 400 times vision field in groups A, B, and C ( F = 81.10, P < 0.01), and the number in group C was lager than that in group A or B ( with P values below 0. 01). Compared with that before treatment, the number of type II macrophages in all the 3 groups was significantly increased (with t values from -69.34 to -47.95, P values below 0.01).

CONCLUSIONSVSD combined with irrigation of oxygen loaded fluid can raise the partial pressure of oxygen of the skin around the wounds effectively, promoting the transition of macrophages from type I to type II, thus it may promote the growth of granulation tissue, resulting in a better recipient for skin grafting or epithelization.

Debridement ; Drainage ; Granulation Tissue ; Humans ; Leg Ulcer ; etiology ; surgery ; Macrophages ; Microvessels ; Negative-Pressure Wound Therapy ; methods ; Nitric Oxide Synthase Type II ; Oxygen ; Skin ; Skin Transplantation ; Skin Ulcer ; Surgical Flaps ; Treatment Outcome ; Vacuum ; Vascular Endothelial Growth Factor A ; Veins ; Wound Healing

0.05).Myocardial perfusion defect scores were decreased significantly from 14.8?3.0 to 10.5?1.8(P

Objective:To explore the role and mechanism of Talin-1 in mouse aortic dissection.Methods:Sixty male FVB mice were evenly divided into groups of blank, model, Talin-1 up-regulation, Talin-1 up-regulation control, Talin-1 down-regulation, and Talin-1 down-regulation control. Except mice in the blank group, mice were treated with β-aminopropionitrile (BAPN) combined with angiotensin to construct a mouse aortic dissection model. Hematoxylin-eosin and vascular elastic fiber staining (EVG) were used to observe the aorta and elastic fiber morphology and structure. Western blot was used to detect the phosphorylation levels of FAK and ERK1 / 2 in mouse aortic tissue.Results:The success rate of aortic dissection in model mice was 70%, and there was no aortic dissection appeared in the blank group.No mice died during the experiment. The incidence of aortic dissection in the Talin-1 down-regulated group was 100%, which was significantly higher than that in the Talin-1 down-regulated control group( P<0.05). The incidence of aortic dissection in the Talin-1 up-regulated group was 20%, significantly lower than that in the Talin-1 up-regulated control group. The wall thickness of the aorta of mice in the Talin-1 down-regulated group was accompanied by hematoma or pseudocavity formation. The median elastic fiber content was higher than that in the Talin-1 downregulation control group( P<0.05). The content of elastic fibers in the blood vessel wall of mice in the Talin-1 up-regulation group was significantly higher than that in the Talin-1 up-regulation control group.The down-regulation of Talin-1 significantly inhibited FAK phosphorylation, and instead promoted ERK1/2 phosphorylation( P<0.05). Conclusions:Down-regulation of Talin-1 may reduce the elastic fiber content in the aorta of mice by activating the ERK1/2 signaling pathway, leading to vascular remodeling of the aortic wall and promoting the occurrence of aortic dissection.

Listeria monocytogenes is a pathogenic bacterium, therefore, it is essential for food safety monitoring to establish a rapid and specific detecting method. In this study, immunomagnetic beads and selective medium were combined to detect Listeria monocytogenes at different concentrations (10(1)-10(5) CFU/mL). Other three types of Listeria spp., Staphylococcus aureus and Vibrio parahaemolyticus were also detected to conduct the cross-reaction analysis. Meanwhile, contaminated milk samples were prepared to explore the limit of detection of immunomagnetic beads combining with selective medium. Results showed that Listeria monocytogenes with the concentration of 10(3) CFU/mL and above was successfully detected. Milk samples were detected within 6 hours, with a detection limit of 0.7 CFU/mL. The method developed is capable of detecting milk samples within 30 h, which is 38 h faster compared with national standard method with the same sensitivity.
Bacteriological Techniques ; methods ; Culture Media ; chemistry ; Immunomagnetic Separation ; methods ; Listeria monocytogenes ; growth & development ; isolation & purification ; Sensitivity and Specificity

In order to study the cytocompatibility of nanophase hydroxyapatite ceramic in vitro, we prepared hydroxyapatite by use of the wet chemistry techniques. The grain size of hydroxyapatite of interest to the present study was determined by scanning electron microscopy and atomic force microscopy with image analysis software. Primary culture of osteoblast from rat calvaria was established. Protein content, synthesis of alkaline phosphatase and deposition of calcium-containing mineral by osteoblasts cultured on nanophase hydroxyapatite ceramics and on conventional hydroxyapatite ceramics for 7, 14, 21 and 28 days were examined. The results showed that the average surface grain size of the nanophase and that of the conventional HA compact formulations was 55 (nanophase) and 780 (conventional) nm, respectively. More importantly, compared to the synthesis of alkaline phosphatase and deposition of calcium-containing mineral by osteoblasts cultured on nanophase was significantly greater than that on conventional ceramics after 21 and 28 days. The cytocompatibility was significantly greater on nanophase HA than on conventional formulations of the same ceramic.
Alkaline Phosphatase ; metabolism ; Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; Cells, Cultured ; Ceramics ; chemistry ; Durapatite ; chemistry ; Microscopy, Atomic Force ; Microscopy, Electron, Scanning ; Nanostructures ; chemistry ; Osteoblasts ; cytology ; Rats

Objective To explore the effect of interleukin-6 (IL-6) and Interleukin-12(IL-12) on immune response to hepatitis B vaccination in infants of HBsAg-positive mothers.Methods A total of 91 neonates whose mothers were HBsAg-positive were included and followed up for 12 months.HBV DNA and HBV serological markers in the peripheral blood of the neonates and infants were detected with fluorescence quantitative polymerase chain reaction (FQ-PCR) and chemiluminescence immunoassay (CLIA),and the levels of IL-6 and IL-12 in the peripheral blood of the neonates and infants were detected with enzyme-linked immunosorbent assay (ELISA).Results The non-/hypo-response rate to hepatitis B vaccination was 35.16％ (32/91) in the 91 infants.In the neonatal period and infantile period,the level of IL-6 in non-/hypo-response group was lower than that in high-response group,while the level of IL-12 was higher than that in high-response group,and there was significant difference (P＜0.01).From the neonatal period to the infantile period,the level of IL-6 increased,while the level of IL-12 descended in both groups,and there was significant difference (P＜0.01).Furthermore,the level of anti-HBs of infants was positively correlated with the level of IL-6 (rs =0.70,0.79,P＜ 0.01),and was negatively correlated with the level of IL-12 (rs=-0.71,-0.72,P＜0.01) in the neonatal period and the infantile period.From the neonatal period to the infantile period,the increased level of IL-6 was positively associated with the level of anti-HBs (rs =-0.74,P＜0.01),while the decreased level of IL-12 was negatively associated with the level of anti-HBs (rs=-0.42,P＜0.01).The level of IL-6 was negatively correlated with the level of IL-12 in the neonatal period and the infantile period (rs=-0.68,-0.70,P＜0.01).Conclusions IL-6 might promote the immune response to hepatitis B vaccination in infants whose mothers were HBsAg-positive,while IL-12 might inhibit the immune response.IL-6 and IL-12 would affect the immune response to hepatitis B vaccination in infants of HBsAg-positive mothers at the same time.

Objective To explore the expression and significance of vitamin D (VitD) in patients with rheumatoid arthritis (RA),and analyze the relationship between its expression and clinical indicators.Methods Clin-ical parameters and laboratory examinations of RA cases (n=250) were collected.Clinical parameters included were gender,age,disease course,swollen joints number,tenderness joints number,visual analog pain score (VAS),disease activity score (DAS)28 score.Laboratory examinations included erythrocyte sedimentation rate (ESR),C-reactive protein (CRP),rheumatoid factor (RF) and anti-cyclic citrullinated peptide (CCP) antibody,antinuclear (ANA) antibody,antikeratin (AKA) antibody,anti-perinuclear factor (APF),anti-mutated citrullinated vimentin (MCV),antibody and anti-6-glucose phosphate isomerase (GPI) antibody,lymphocyte subsets in the peripheral blood and lymphocyte subsets of CD4+T cells.The level of 25-(OH)-Vit-D and clinical parameters,laboratory examinations were analyzed retrospectively.One-way ANOVA and KruskalWallis test were used for comparison among the groups;and the correlation analysis was performed by Pearson and Spearman rank correlation analysis.Results ① The level of 25-(OH) D in RA patients was significantly lower than that in healthy controls (t=11.676,P＜0.01).② According to 25-(OH)D level,RA patients were divided into the deficiency group,insufficient group and normal group,the tender joints count (x2=17.793,P＜0.001),the number of swollen joints (x2=12.635,P=0.002),ESR (F=6.330,P=0.002),VAS score (F=5.095,P=0.007,DAS28 (F=4.990,P=0.008) were different significantly amorg the three groups.③RF (x2=6.742,P=0.034) and anti-CCP antibody (x2=6.836,P=0.033) were different significantly among the three groups and the level of 25-(OH) D was negatively correlated with RF (r=-0.202,P=0.001),anti-CCP antibody (r=-0.220,P＜0.01),anti-MCV antibody (r=-0.109,P=0.002) and AKA (r=-0.215,P=0.001).④ The level of 25-(OH) D in the RF (t=-2.715,P=0.007),anti-CCP antibody (t=-2.03,P=0.044),AKA (t=-2.108,P=0.036) negative group was significantly higher than that in patients with antibody positive group.⑤ The level of Th1 (IFN-γ) cells (F=3.259,P=0.043) and Treg (CD4+CD25+Foxp3+) cells (F=4.342,P=0.031) were significantly different among the three groups and the level of 25-(OH) D was positively correlated with Treg (CD4+CD25+Foxp3+) cells (r=0.146,P=0.025).Conclusion Vitamin D is generally deficient in RA patients,which is significantly correlated with disease activity,RF,anti-CCP antibody,anti-MCV antibody,AKA and Th1,Treg cells.It is suggested that vitamin D may play an important role in the immunological pathogenesis and disease progression of RA.