BACKGROUND:Preparation of titanium/hydroxyapatite composite by conventional methods has the deficiency of simple structure, low degree of automation and difficulty in porosity and pore size control, which limits the diverse process and manufacture. OBJECTIVE:To evaluate the feasibility of three-dimensional printing technology for the preparation of titanium/hydroxyapatite composite and titanium/hydroxyapatite functional y graded material molding. METHODS:A CAD model of titanium/hydroxyapatite composite was designed to be the cylinder (diameter 25 mm, height 20 mm), while the titanium/hydroxyapatite functional y graded implant designed as a CAD model of the cylinder with 25 mm in diameter asnd 10 mm in height with two layers, the upper layer with titanium powder and the lower layer with titanium/hydroxyapatite powder. The composite and functional y graded implant were processed by the three-dimensional printing and sintered. The sintered titanium/hydroxyapatite composite and titanium/hydroxyapatite functional y graded implant were observed for their microstructures, and the X-ray diffraction analysis and compressive strength testing were performed. RESULTS AND CONCLUSION:The sintered titanium/hydroxyapatite composite and titanium/hydroxyapatite functional y graded implant had uniform contraction and no obvious distortion. The sintered titanium/hydroxyapatite composite had the aperture size from 50 to 150μm. There occurred a chemical reaction between titanium and hydroxyapatite during the sintering process, obtaining the new creations of Ca3(PO4)2, CaTiO3, TiO2 and CaO. Its compressive strength was (184.3±27.1) MPa. The microstructure of titanium/hydroxyapatite functional y graded implant had graded structures with a visible line between the two layers. The results of the microstructure and mechanical properties of titanium/hydroxyapatite composite and titanium/hydroxyapatite functional y graded implant can meet the requirements of medical biological implant materials.

13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.

To study the value of lung ultrasound score (LUS) in assessing the clinical outcome of patients with ventilator-associated pneumonia (VAP).A total of 99 VAP patients were enrolled in a prospective study.All patients met the diagnostic criterion of VAP based on the 2013 guidelines and admitted into our ICU from Jun 2013 to Jun 2015.All parameters were recorded on the diagnostic day (day 1) and day 5,including LUS,clinical pulmonary infection score (CPIS),chest X ray (CXR),Acute Physiology and Chronic Health Evaluation Ⅱ (APACHE Ⅱ) score,Sequential Organ Failure Assessment (SOFA) score,etc.According to the CPIS,patients were divided into 2 groups(CPIS less than 6 and more or equal to 6).CPIS and LUS were similar on day 1 between two groups (P ＞ 0.05).However,on day 5,significant differences of CPIS and LUS were found between groups with CPIS ＜ 6 and CPIS≥6 (P =0.019 and P ＜ 0.001 respectively).LUS decreased on day 5 in CPIS ＜ 6 group and increased in CPIS ≥6 group.In CPIS ＜ 6 group,there was a positive correlation between LUS and CPIS on day 1 (r =0.375,P =0.003) and day 5 (r =0.590,P ＜ 0.001).CPIS ≥6 groupshowed the same trend on day 1 (r =0.484,P =0.002) and day 5 (r =0.407,P =0.011).LUS can be used to dynamically evaluate the clinical outcome of VAP.

The chemical constituents of Safflower injection were isolated and purified by polyamide, silica gel, Sephadex LH-20, ODS column chromatographies and preparative HPLC. As a result, sixteen compounds have been isolated. Based on the spectral data analysis, their structures were elucidated as scutellarin (1), kaempferol-3-O-β-rutinoside(2), hydroxysafflor yellow A(3), rutin (4), coumalic acid(5), adenosine(6), syringoside(7), (3E)-4-(4'-hydroxyphenyl)-3-buten-2-one(8), (8Z)-decaene-4, 6-diyne-1-Oβ-D-glucopyranoside(9), 4-hydroxybenzaldehyde (10), (2E, 8E) -tetradecadiene-4, 6-diyne-1, 12, 14-triol-1-O-β-D-glucopyranoside (11), kaem-pferol-3-O-β-sophorose (12), uridine (13), roseoside (14), cinnamic acid (15), and kaempferol (16). Compounds 1,2,7,9,11 and 12 were isolated from the Safflower injection for the first time. The anti-platelet aggregation activities of the isolated compounds were assayed. The results indicated all tested compounds exhibited potent activity except for 5, while 2, 3, 9 and 12 showed strong activity against platelet aggregation.
Animals ; Blood Platelets ; drug effects ; physiology ; Carthamus tinctorius ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Fibrinolytic Agents ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Platelet Aggregation ; drug effects ; Rabbits ; Spectrometry, Mass, Electrospray Ionization

ObjectiveTo evaluate the value of lung ultrasound score (LUS) on assessing the severity and prognosis in patients with acute respiratory distress syndrome (ARDS), and to investigate its correlation with oxygenation index, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score, sequential organ failure assessment (SOFA) score, and clinical pulmonary infection score (CPIS), and other traditional parameters.Methods A prospective double-blind cohort study was conducted. Sixty-two ARDS patients conformed to the Berlin diagnostic criteria admitted to intensive care unit (ICU) of Beijing Huaxin Hospital from October 2013 to December 2014 were enrolled, including 14 cases with mild, 18 moderate, and 30 severe ARDS; among them 37 cases were of ARDS with pulmonary origin, and 25 non-pulmonary ARDS; 35 patients survived, and 27 died. The clinical data and scores of all patients were recorded by one specialized observer, including baseline data, hemodynamic parameters, lactate, respiratory parameters, and APACHEⅡ, SOFA and CPIS scores. Another observer of recording was responsible for the results of lung ultrasound, LUS, and echocardiogram. The correlation between LUS and oxygenation index as well as APACHEⅡ, SOFA and CPIS scores was analyzed by bivariate correlation analysis. Receiver operator characteristic curve (ROC) was plotted, and the predictive value, sensitivity and specificity of mild ARDS, moderate ARDS, severe ARDS and mortality by LUS were calculated. Results LUS had a negative correlation with oxygenation index (r = -0.755,P< 0.001), a good positive correlation with APACHEⅡ (r = 0.504,P< 0.001), SOFA (r = 0.461,P< 0.001) and CPIS (r = 0.571,P< 0.001) was found. LUS in the pulmonary ARDS group had a positive correlation with CPIS (r = 0.399,P< 0.05), and a positive correlation was found in non-pulmonary ARDS group (r = 0.350,P< 0.05), which indicated that the correlation in pulmonary ARDS was more satisfactory than that in non-pulmonary ARDS. LUS in the pulmonary ARDS group was significantly higher than that in non-pulmonary ARDS group (22.1±4.9 vs. 11.3±2.1,t = 11.667,P< 0.001); LUS in mild, moderate, severe ARDS groups was 9.9±1.7, 14.0±1.4, 23.6±4.1. The predictive value for mild ARDS by LUS was 7.0, sensitivity of 87.0%, specificity of 89.0%; that for moderate ARDS was 11.0, sensitivity of 89.0%, specificity of 87.0%; that for severe ARDS was 8.0, sensitivity of 90.0%, specificity of 88.5%. LUS was 24.3±3.8 in the death group, and 12.7±2.9 in the survival group. Area under ROC curve (AUC) was calculated, and the patients with LUS> 19.0 had a high mortality, sensitivity for predicting death was 84.0%, and specificity of 89.0%.Conclusion Bedside LUS, which is simple and easily available, could evaluate the changes in pulmonary ventilation area of ARDS, and its degree of severity, and prognosis including prediction of mortality of the patients.

ObjectiveTo evaluate the application of a whole blood interferon-γ (IFN-γ) release assay QuantiFERON-TB Gold In Tube (QFT-GIT) in the diagnosis of tuberculous pleural effusion.Methods IFN-γ released by specific T cells stimulated by early secreted antigenic target 6 × 103protein (ESAT-6),culture filtrate protein 10 (CFP -10) and TB7.7 were measured by QFT-GIT test in 44 tuberculous pleural effusion patients and 16 non-tuberculous pleurisy controls.The IFN-γ release level between groups was compared by Mann-Whitmey test.ResultsThe positive rates of QFT-GIT in patients with tuberculous pleural effusion and non tuberculous pleurisy were 95.5％ and 12.5％,respectively.The sensitivity,specificity,positive predictive value and negative predictive value of QFT-GIT were 95.6％,87.5％,95.6％ and 87.5％,respectively.The antigen-specific IFN-γ release level in the patients with tuberculous pleural effusion was significantly higher than that in non-tuberculous pleurisy controls (P＜0.01).Conclusions The whole blood INF-γ release assay QFT-GIT is a sensitive and specific assay for detecting pleural tuberculosis infection.It could be a useful diagnostic tool for the diagnosis of tuberculous pleural effusion in China.

Transient forebrain ischemia-reperfusion(I/R);Brain-derived neurotrophic factor(BDNF);Histone deacetylase 3(HDAC3);Hippocampus Objective To evaluate the effect of transient forebrain ischemia-reperfusion(I/R)on the binding of brainderived neurotrophic factor(BDNF)promoters to histone deacetylase 3(HDAC3)in the hippocampus of rat and investigate its mechanism.Methods The I/R model of SD rats(I/R group)was established by Pulsinelli four-vessel clamping method,and sham operation group(Sham group)was set at the same time,which were observed for the survival of neurons in the hippocampus of rats by Nissl staining,detected for the binding of BDNF promoters(Bdnf-p1,Bdnf-p2,Bdnf-p4 and Bdnf-p6)to HDAC3 by chromatin immunoprecipitation(ChIP)and determined for the expression of brain derived neurotrophic factor antisense(BDNF-AS)by qPCR.Results Compared with Sham group,the quantity of neurons in hippocampal CA1 region of rats decreased significantly in I/R group,while those in CA3 region and DG region showed no significant changes.The binding levels of Bdnf-p1 and Bdnf-p2 to HDAC3 in hippocampal CA1 region decreased significantly in I/R Group(t = 2.575 and 2.241 respectively,each P<0.05),while there was no significant difference in the binding levels of Bdnf-p4 and Bdnf-p6 to HDAC3(t = 1.033 and 0.348 respectively,each P>0.05);The binding levels of Bdnf-p1 and Bdnf-p2 to HDAC3 in CA3 region increased significantly(t = 12.600 and 3.191,P<0.001 and<0.05,respectively),while the binding level of Bdnf-p6 to HDAC3 decreased significantly(t = 4.029,P<0.05)and no significant difference was observed in the binding level of Bdnf-p4 to HDAC3(t = 0.175,P>0.05);In DG region,the binding level of each BDNF promoter to HDAC3 showed no significantly difference(t = 0.630 ～ 1.687,each P>0.05).Meanwhile,the expression level of BDNF-AS in hippocampal CA1 region of rats decreased significantly(t = 2.560,P<0.05),but increased significantly in hippocampal CA3 and DG regions(t = 3.543 and 3.637 respectively,each P<0.01)in I/R group.Conclusion I/R showed a significant effect on the binding level of BDNF promoter to HDAC3 in rat hippocampus,which may play a role by changing the expression level of BDNF-AS.

OBJECTIVETo construct, express and identify the recombinant plasmid pcDNA3.1(+)/sflk1-IFN-gamma encoding bifunctional protein sflk1-IFN-gamma (soluble fetal liver kinase 1 and interferon-gamma).

METHODSsflk1 and IFN-gamma gene fragments were cloned by RT-PCR, and then inserted into pcDNA3.1(+) plasmid between BamHI-EcoRI and XhoI-XbaI restriction sites to form the recombinant plasmid pcDNA3.1(+)/sflk1-IFN-gamma. The recombinant sflk1-IFN-gamma transiently expressed in COS-7 cells was detected by ELISA and Western blotting. Bioactivities of sflk1-IFN-gamma fusion protein were identified by proliferation inhibition assay with H5V cells and NK activity assay.

RESULTSpcDNA3.1(+)/sflk1-IFN-gamma can be effectively expressed in COS-7 cells. Concentrations of sflk1 and IFN-gamma in culture supernatants of pcDNA3.1(+)/sflk1-IFN-gamma transfected COS-7 cells were (20.85+/-2.48) ng/ml and (1.08+/-0.09) ng/ml, respectively. Western blotting showed that the molecular weight of sflk1-IFN-gamma fusion protein was about 130 kDa, while that of sflk1 was 115 kDa. The supernatants of transfected cells significantly inhibited the proliferation of H5V cells stimulated by mouse VEGF 164 and enhanced the NK activity of splenocytes, demonstrating that sflk1-IFN-gamma fusion protein possessed the bioactivities of both sflk1 and IFN-gamma.

CONCLUSIONThe constructed plasmid pcDNA3.1(+)/sflk1-IFN-gamma can be effectively expressed in eukaryotes. The expressed sflk1-IFN-gamma fusion protein has the biological activities of both sflk1 and IFN-gamma.

Animals ; COS Cells ; Cercopithecus aethiops ; Female ; Interferon-gamma ; biosynthesis ; genetics ; Mice ; Mice, Inbred C57BL ; Plasmids ; Recombinant Proteins ; analysis ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Vascular Endothelial Growth Factor Receptor-2 ; biosynthesis ; genetics

Aim To investigate the effect of TaorenHonghua drug pair on intervertebral disc degeneration (IVDD) in rats.Methods Fifty healthy Wistar rats were randomly divided into control group,model group,sham group,meloxicam group and Taoren-Honghua drug pair group,with 10 rats in each group.We established dynamic and static forces imbalance of cervical disc degeneration model or sham surgery in rats.12 weeks later,rats were intragastrically administered with meloxicam,Taoren-Honghua drug pair or saline for 30 days.C4/5 and C6/7 discs were harvested from rats.ABOG staining was used for observation of intervertebral disc morphology,real time PCR for mRNA expressions of type Ⅱ collagen (Col Ⅱ) and type Ⅹ collagen (Col Ⅹ),and immunohistochemical staining for Col Ⅱ and Col Ⅹ.Results Compared with model group,Col Ⅱ expression increased,while Col X expression decreased in chondrocyte of intervertebral disc in Taoren-Honghua-treated group(P ＜ 0.01).Conclusion Taoren-Honghua drug pair could delay the degeneration of cartilage endplate in rat intervertebral disc.

OBJECTIVETo investigate the long-term integrity and the biological function of interface between the bioadhesive peptide modified implant surface and peri-implant tissue.

METHODSA short bioadhesive peptide containing Glycine-Tyrosine-Arginine-Glycine-Asparticacid-Serine (GYRGDS) sequence was immobilized onto the titanium implant surface by means of sol-gel coating technique and self-assembled monolayers (SAM). The chemical composition and organic functional groups on the titanium surfaces were characterized using XPS (X-ray photoelectron spectroscopy) and FTIR (Fourier transform infrared spectrometer). The adhesive strength and stability of osteoblasts on various implant surfaces were compared under flow condition.

RESULTSThe results showed that alkali/hot water aging treatment could apparently improve the content of -OH functional groups of titanium surface. The chemical reactive Ti-O-Ti bonding at the surface of titanium played a vital role in inducing the formation of organosilane SAM. GYRGDS peptide can be covalently grafted onto the surface of titanium by SAM technique. The resistance of freshly adherent osteoblasts to detachment by flow was shear time dependent. When the four groups were compared under the same flow stress condition (2.05 Pa) at three specific time spans (30 min, 1 h, 2 h), the cells retention rates in GYRGDS-grafted groups were 93.0%, 54.4%, 34.4% respectively and were much higher than those in non-coated groups.

CONCLUSIONSIt was suggested that GYRGDS might have positive effects on maintaining stability and adherence of cells onto the substrates under flow condition.

Dental Cements ; Osteoblasts ; physiology ; Peptides ; pharmacology ; Prostheses and Implants ; Surface Properties ; Titanium