BACKGROUND: Microarc oxidation (MAO) is a break-through anodyzing technology for forming oxide films on valve metal.Use of this technology allows thick, porous oxide layers to be formed on the surface of pure titanium. Few biocompatibility reports using this treatment have been found.OBJECTIVE: The blood compatibility of a novel surface modified titanium substrata biomaterial using MAO was investigated.DESIGN: Positive and negative control, contrast observation and gold standard control.SETTING: Wuhan Union Hospital.MATERIALS: A healthy male adult New-Zealand rabbit, weighing 2.5 kg and ordinary grade, was selected in this study.Pure titanium sticks TA1 (Baoji Yingnaite Non-ferrous Metal Co., Ltd.), MAO-Ti and 20 g/L potassium oxalate were also selected in this study.METHODS: The study was carried out in the Laboratory of General Surgery, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology in May 2006. ① Materials: Titanium substrate of 10 mm in diameter and 2 mm in depth was put in an electrolyte which was quipped with deionized water, dibasic sodium phosphate, and ethanoic acid calcium for MAO treatment for 10 minutes. ② Groups: Three groups were analysed: test group, negative control group and positive control group. Test group: MAO-Ti was dipped in 10 mL saline; Positive control group: 10 mL deionized water was added in each tube; Negative control group: 10 mL saline was added in each tube. ③ Operation: Fresh whole blood was collected from rabbit and then mixed with the liquids in the three groups respectively after anti-coagulation. In addition, UV-Visible Spectrophotometer was used to evaluate the hemolytic ratio. A hemolytic ratio below or equal to 5% indicated that this novel material fitted the requirements. On the contrary, a hemolytic ratio higher than 5% proofed the existence of a hemolyzation.MAIN OUTCOME MEASURES: The hemolytic ratio of materials in three groups.RESULTS: The hemolytic ratio of the test group was 0.90%. The result indicated that this new material had no haemolysis effect.CONCLUSION: The material does not resolve red blood cells and is coincident with the international and governmental standard.

Objective To investigate the dynamic changes of microglial polarization at the perihe-matoma area and provide timepoint evidence for interventing microglial polarization as well as studying the polarization mechanism after intracerebral hemorrhage ( ICH ) . Methods Healthy male Sprague Dawley (SD) rats were randomly divided into sham group,ICH-4 h,1 d,3 d,7 d and 14 d groups with 6 in each group. The rats in ICH groups were injected collagenase VII-s into the caudate nucleus to establish the in-tracerebral hematoma model and rats in sham operated group were treated with the same amount of saline. The brains were taken at 4 h,1 d,3 d,7 d,14 d in the ICH group,1 d in sham group. Microglia typeⅠ( M1, CD11b++CD86+) and microglia typeⅡ( M2,CD11b++Arg-1+) were examined by immunofluorescence and the number of M1 and M2 around hematoma were analyzed. Results ( 1) The M1 and M2 were both ob-served at 4 h after ICH and a small quantity of branches were still presented on M1. ( 2) M1 took the main position in acute stage (1~3 d),early subacute stage(3~7 d) and chronic stage (>14 d) after ICH.The number of M2 was elevated transiently in superacute (<24 h) and late subacute stage (7 d).The number of M2 (31.40±1.69) was more than M1 (21.43±1.81) at 4 h after ICH ( t=- 4.085, P=0.002),and the number of M2 (116.25±5.06) significantly exceeded M1 (85.75±7.32) again on day 7 ( t=-0.690, P=0.001). Conclusion M1 is in a dominant position in acute,early subacute and chronic stages after ICH;M2 is dominant in superacute and late subacute stages. Investigating the mechanism of M2 formation at acute period ( such as 4 h) or late subacute stage ( such as 7 d) ,and inhibiting M1 formation in the early subacute stage ( 1~3 d) have important significance for clinical treatment of ICH.

Objective To assess under different vessel diameter ,the effect of the aspiration thrombectomy catheter in improving the myocardial reperfusion and clinical prognosis in patients with acute myocardial infarction (AMI)who were undergone primary percutaneous coronary intervention(PCI) .Methods 205 patients with AMI immediate implant stents after thrombus suction ,the TIMI flow grade(myocardial infarction thrombolysis treatment test flow classification ) ,postoperative ecg evolution ,incidence of no-reflow MACE in 30 days and MACE in 6 months were compared between conventional thrombus suction group and suction again group(blood vessels of <3 .0 mm and ≥3 .0 mm) .Results The level 3 blood flow rate ,MACE in 6 months in suction again group with blood vessels of ≥3 .0 mm had improved significantly ,but had no beneficial effects in blood vessels of ≥3 .0 mm .Conclusion In AMI patients treated with primary PCI ,application of aspiration thrombectomy catheter with blood vessels of ≥3 .0 mm may im-prove the flow condition before infarction related blood vessels ,reduce MACE .

OBJECTIVETo investigate effects of low-dose cyclophosphamide and prednisone (CP) metronomic chemotherapy on microvessel density of bone marrow, serum vascular endothelial growth factor (VEGF) and platelet derived growth factor BB (PDGF-BB)in multiple myeloma (MM) patients.

METHODS54 refractory or relapsed MM patients were treated with CP metronomic chemotherapy consisted of oral cyclophosphamide (CTX, 50 mg/d) and prednisone (Pred, 15 mg/d). Bone marrow and peripheral blood of each patient were collected before and 2, 4, 6 months after treatment. Among the 37 assessable patients, 30 cases were responsive with the response rate of 81.08%. Another 17 cases were follow-uped less than 6 months or failure to obtain serum samples or lost to follow-up. Microvessel density of bone marrow was measured by immunohistochemistry and serum VEGF/PDGF-BB expression was analyzed by ELISA in the 37 assessable patients.

RESULTS2, 4, 6 months following CP metronomic chemotherapy, microvessel densities of bone marrow in the responders were 33.1 ± 4.8/HP, 24.8 ± 3.7/HP, 19.7 ± 2.1/HP respectively; the expressions of VEGF were (394 ± 57) ng/L, (268 ± 32) ng/L and (217 ± 20) ng/L respectively; the expressions of PDGF-BB were (304 ± 31) ng/L, (274 ± 31) ng/L and (196 ± 22) ng/L respectively. After CP metronomic chemotherapy, there were significantly lower of microvessel density, VEGF and PDGF-BB levels than pretreatment \[MVD 48.5 ± 5.9/HP, VEGF (517 ± 60) ng/L, PDGF-BB (484 ± 60) ng/L\]in the responders (P < 0.01). While in the non-responders, after treated by CP metronomic chemotherapy for 2 months, microvessel density, the expression of VEGF and the expression of PDGF-BB were 32.5 ± 4.7/HP, 512 ± 39 ng/L and (452 ± 39) ng/L respectively. There were no significant changes of MVD, VEGF and PDGF-BB levels compared with pretreatment \[MVD 33.2 ± 5.6/HP,VEGF (498 ± 55) ng/L, PDGF-BB (488 ± 44) ng/L\] (P > 0.05).

CONCLUSIONSOur findings suggested that continuous low-dose CP metronomic chemotherapy could decrease microvessel density of bone marrow in MM patients. Furthermore, it down-regulated expression of serum VEGF and PDGF-BB to exert its anti-angiogenesis in MM.

Aged ; Aged, 80 and over ; Angiogenesis Inhibitors ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclophosphamide ; administration & dosage ; Female ; Humans ; Male ; Microvessels ; drug effects ; Middle Aged ; Multiple Myeloma ; blood ; blood supply ; drug therapy ; Prednisone ; administration & dosage ; Proto-Oncogene Proteins c-sis ; blood ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVETo find a new method for edge detection and modeling in frontal facial contour image.

METHODSSearching circle-based edge detection algorithm was developed on the basis of Sobel edge detector. Apriori knowledge of the facial contour and searching limitations as the minimum curvature radius, concave-convex property, and maximum edge disconnected distance were used to detect the edge of frontal facial contour. The frontal facial contour model was established with least squares curve fitting methods, and the relationship between the model rank and model precision was analyzed.

RESULTSThe edge detected by the new method was consistent with the actual edge of the facial contour and the irrelevant edge was well eliminated. Variation of the 2,4,10 time models from the actual image were compared, which identified the 10 time model as the best one.

CONCLUSIONThe effectiveness and practicability of this new method for edge detection and modeling has been tested, which provides a theoretic basis for designing facial contouring image system.

Algorithms ; Computer Simulation ; Face ; anatomy & histology ; Humans ; Image Interpretation, Computer-Assisted ; methods ; Models, Anatomic ; Pattern Recognition, Automated ; methods ; Reproducibility of Results ; Surgery, Plastic ; methods

OBJECTIVETo investigate the association of single nucleotide polymorphisms (SNPs) of the peptidylarginine deiminase IV (PADI4) and HLA-DRB1 shared epitope (SE) alleles with rheumatoid arthritis(RA) in a Chinese population.

METHODSFour exonic SNPs of the PADI4 gene (PADI 4_89*A/G, PADI 4_90*C/T, PADI 4_92*C/G and PADI 4_104*C/T) were genotyped in 67 unrelated patients with RA and 81 healthy controls, using cDNA sequencing and T vector cloning. HLA-DRB 1*01, *04 and *10 subtypes were determined by polymerase chain reaction with sequence specific primers (PCR-SSP).

RESULTSThe distributions of the 4 SNPs were different in the two groups, and increased RA susceptibility was significantly associated with the minor alleles of PADI 4_89*G (P was 0.023), PADI 4_90*T (P was 0.004), PADI 4_104*T (P was 0.003), and the haplotypes carrying the 4 minor alleles (P was 0.008). HLA-DRB1 SE alleles are composed of HLA-DRB 1*0101, *0102, *0401, *0404, *0405, *0408, *0409, *0410 and *1001. Individuals carrying the SE alleles were associated with increased RA susceptibility (P was 0.002). Individuals carrying both the SE alleles and minor alleles of the 4 SNPs were more susceptible to RA than individuals carrying neither the minor SNP alleles nor the SE alleles.

CONCLUSIONThe PADI4 SNPs and haplotypes are associated with RA susceptibility in Chinese. HLA-DRB1 shared epitope is also an important risky factor for RA. There may exist certain synergistic effect between the PADI4 minor alleles and the HLA-DRB1 shared epitope.

Adult ; Aged ; Aged, 80 and over ; Alleles ; Arthritis, Rheumatoid ; genetics ; Asian Continental Ancestry Group ; genetics ; Case-Control Studies ; Epitopes ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; HLA-DR Antigens ; genetics ; HLA-DRB1 Chains ; Humans ; Hydrolases ; genetics ; Male ; Middle Aged ; Phenotype ; Polymorphism, Single Nucleotide ; Protein-Arginine Deiminases

The inherent evolvability of promiscuous enzymes endows them with great potential to be artificially evolved for novel functions. Previously, we succeeded in transforming a promiscuous acylaminoacyl peptidase (apAAP) from the hyperthermophilic archaeon Aeropyrum pernix K1 into a specific carboxylesterase by making a single mutation. In order to fulfill the urgent requirement of thermostable lipolytic enzymes, in this paper we describe how the substrate preference of apAAP can be further changed from p-nitrophenyl caprylate (pNP-C8) to p-nitrophenyl laurate (pNP-C12) by protein and solvent engineering. After one round of directed evolution and subsequent saturation mutagenesis at selected residues in the active site, three variants with enhanced activity towards pNP-C12 were identified. Additionally, a combined mutant W474V/F488G/R526V/T560W was generated, which had the highest catalytic efficiency (k (cat)/K (m)) for pNP-C12, about 71-fold higher than the wild type. Its activity was further increased by solvent engineering, resulting in an activity enhancement of 280-fold compared with the wild type in the presence of 30% DMSO. The structural basis for the improved activity was studied by substrate docking and molecular dynamics simulation. It was revealed that W474V and F488G mutations caused a significant change in the geometry of the active center, which may facilitate binding and subsequent hydrolysis of bulky substrates. In conclusion, the combination of protein and solvent engineering may be an effective approach to improve the activities of promiscuous enzymes and could be used to create naturally rare hyperthermophilic enzymes.
Aeropyrum ; chemistry ; enzymology ; Archaeal Proteins ; genetics ; metabolism ; Binding Sites ; Biocatalysis ; Caprylates ; metabolism ; Cloning, Molecular ; Dimethyl Sulfoxide ; chemistry ; Escherichia coli ; Hot Temperature ; Industrial Microbiology ; methods ; Kinetics ; Laurates ; metabolism ; Molecular Dynamics Simulation ; Mutagenesis, Site-Directed ; methods ; Peptide Hydrolases ; genetics ; metabolism ; Protein Binding ; Protein Conformation ; Recombinant Proteins ; genetics ; metabolism ; Solvents ; chemistry ; Substrate Specificity

OBJECTIVETo investigate the effects of T-2 toxin on expressions of Fas, p53, Bcl-xL, Bcl-2, Bax and caspase-3 on human chondrocytes.

METHODSHuman chondrocytes were treated with T-2 toxin (1-20 ng/ml) for 5 d. Fas, p53 and other apoptosis-related proteins such as Bax, Bcl-2, Bcl-xL, caspase-3 were determined by Western blot analysis and their mRNA expressions were determined by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIncreases in Fas, p53 and the pro-apoptotic factor Bax protein and mRNA expressions and a decrease of the anti-apoptotic factor Bcl-xL were observed in a dose-dependent manner after exposures to 1-20 ng/ml T-2 toxin, while the expression of the anti-apoptotic factor Bcl-2 was unchanged. Meanwhile, T-2 toxin could also up-regulate the expressions of both pro-caspase-3 and caspase-3 in a dose-dependent manner.

CONCLUSIONThese data suggest a possible underlying molecular mechanism for T-2 toxin to induce the apoptosis signaling pathway in human chondrocytes by regulation of apoptosis-related proteins.

Apoptosis ; drug effects ; Base Sequence ; Blotting, Western ; Caspase 3 ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Cells, Cultured ; Chondrocytes ; cytology ; drug effects ; metabolism ; DNA Primers ; genetics ; Gene Expression ; drug effects ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; T-2 Toxin ; toxicity ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; bcl-2-Associated X Protein ; genetics ; metabolism ; bcl-X Protein ; genetics ; metabolism ; fas Receptor ; genetics ; metabolism

OBJECTIVETo investigate the binding activity of activator protein-1 (AP-1) with DNA probe in the colorectal carcinoma (CRC) tissues and surrounding tissues and explore the correlation between the activation of AP-1 signal transduction pathway and metastasis of CRC.

METHODSThe AP-1 DNA binding activities were investigated by electrophoretic mobility shift assay (EMSA) in CRC specimens (T), surrounding tissues including 2 cm (P(2)), 5 cm(P(5)) far away from primary tumor margin and distal resection margin of the specimens (N). The mRNA expression level of vascular endothelial growth factor (VEGF), matrix metalloproteinase-9 (MMP-9) were measured by quantitive reverse transcription polymerase chain reaction (Q- RT-PCR).

RESULTSThe AP-1 DNA binding activity in T was significantly higher than those in P(2), P(5) and N (P< 0.05) tissues. There were significantly positive correlations between AP-1 DNA binding activity in tumor and invasive degree, lymphatic metastasis respectively (P< 0.01), but no correlation with histological classification and differentiation (P> 0.05). The transcription levels of VEGF and MMP-9 in CRC were significantly higher than those in P(5) and N (P< 0.01, P< 0.05) tissues. The transcription levels of VEGF and MMP-9 were significantly correlated with increasing AP-1 DNA binding activity (P< 0.01).

CONCLUSIONSAP-1 is significantly correlated to the invasion and metastasis in CRC. The activation of AP-1 signal transduction pathway might be involved in the angiogenesis and of degradation extracellular matrix during tumor metastasis.

Colorectal Neoplasms ; metabolism ; pathology ; Female ; Humans ; Lymphatic Metastasis ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Neoplasm Staging ; Neovascularization, Pathologic ; Signal Transduction ; Transcription Factor AP-1 ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo evaluate the clinical value of retrospective electrocardiogram (ECG) -editing technique in dual-source computed tomography (CT) coronary angiography in patients with arrhythmia.

METHODSTotally 73 patients with arrhythmia during dual-source CT coronary angiography were included into this study. A retrospective gating technique and ECG-editing technique (Insert Sync; Disable Sync; Delete Sync; Shift R-peak) were used in patients who needed ECG-editing. Two experienced radiologists evaluated in consensus all pre-editing and post-editing reconstructed images and recorded scores according to the American Heart Association guidelines on coronary segmentation on a per segment basis. Image quality of all coronary segments was assessed using a four-point grading scale from excellent (4 scores) to non-assessable (1 score) .

RESULTSThe overall mean image quality of 34 patients who did not need ECG-editing was 3.42 ± 0.20. In 39 patients who needed ECG-editing, the overall mean image quality before and after ECG-editing was 2.39?0.37 and 3.22?0.24. The mean image quality in every segment between pre-editing and post-editing was also significantly different (P<0.01) .

CONCLUSIONECG-editing technique can remarkably improve image quality of coronary artery segments by reducing or even eliminating the artifact produced by arrhythmia during dual-source CT coronary angiography.

Adult ; Aged ; Aged, 80 and over ; Arrhythmias, Cardiac ; diagnostic imaging ; Coronary Angiography ; methods ; Electrocardiography ; Female ; Humans ; Image Processing, Computer-Assisted ; methods ; Male ; Middle Aged ; Tomography, X-Ray Computed ; methods