Objective To develop an infusion joint of the two-cavity catheter used for deep vein thrombolysis and explore its clinical application.Methods The joint was composed of a perfusion tube,a perfusion tube tee,two connecting hoses,two connecting hard tubes,two Luer tapers,a liquid inlet connected with the tee,No.1 and No.2 liquid outlets.The liquid medicine was driven to flow through the tube and inlet by gravity,and then came into No.1 and No.2 liquid outlets respectively.Results There were no significant differences between the joint and the traditional way when used for deep vein thrombolysis with a two-cavity catheter (P＞0.05),while the joint had patient satisfaction increased significantly and the time consumed decreased obviously when compared with the traditional way (P＜0.05).Conclusion The joint executes deep vein thrombolysis with a two-cavity catheter and only a set of infusion set,gains advantages in simple structure,low cost and easy operation,and thus is worthy promoting clinically.

Adult ; Aged ; External Fixators ; Female ; Fracture Fixation, Internal ; methods ; Fractures, Comminuted ; surgery ; Humans ; Male ; Middle Aged ; Radius Fractures ; surgery

AIMTo investigate the role of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) on pulmonary vascular structural remodeling in rats and pika.

METHODSThe Wistar rats which reside at 2 260 m were carried to 3 417 m. After they were fed 24 hours,2 weeks and 3 weeks respectively, the level of VEGF and ET-1 were measured using a kit by ELISA method. Pulmonary tissue was taken out to stain with elastica-Van Gieson. The amount of pulmonary arteries (< 100 microm) and the component ratio of MA, PMA,and NMA were calculated by using a light microscope. The ratio of right ventricle weight to left ventricle plus septum weight (RV/LV + S) were measured.

RESULTSThe ET-1 was significantly different in pika as compared with 24 h, 2 weeks, 3 weeks hypoxic rats (P < 0.01) respectively. The levels of VEGF in 2 weeks, 3 weeks rats were much higher than that of pika but no difference was found between pika and 24 h hypoxic rats. The ratio of MA, PMA obviously increased, and NMA decreased significantly, right ventricular hypertrophy was developed in differ groups of hypoxic rats.

CONCLUSIONThe VEGF and ET-1 participate the muscularization of pulmonary vessels during hypoxia and play an important role in the process of hypoxic pulmonary hypertension in rats, however the VEGF and ET-1 may be maintainable only normal organic function in pika.

Animals ; Endothelin-1 ; metabolism ; Female ; Hypertension, Pulmonary ; metabolism ; Hypoxia ; metabolism ; Lagomorpha ; Lung ; blood supply ; Male ; Pulmonary Artery ; metabolism ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the developmental profiles on surface expression and co-localization of NMDA receptor clusters and AMPA receptor clusters on dendrite in cultured hippocampal neurons of rats.

METHODSGreen fluorescent protein tagged GluR2 subunit (GFP-GluR2) and FLAG tagged NR2B subunit (FLAG-NR2B) were transfected into cultured hippocampal neurons at 5 days in vitro (DIV5). FLAG-NR2B containing NMDA receptor clusters and GFP-GluR2 containing AMPA receptor clusters expressed on membrane surface were then labeled in living neurons using anti FLAG mAb/Cy3-conjugated anti-mouse antibody and anti-GFP pAb/Alex488-conjugated anti-rabbit antibody.

RESULTThe numbers of receptor cluster per 100 microm dendrite in the neurons at DIV7 and DIV19 were 39.7+/-5.0 and 64.7+/-6.1 (P<0.01) for NR2B-NMDAR, 59.1+/-3.3 and 99.7+/-6.4 (P<0.01) for GluR2-AMPAR, and 29.9+/-4.5 and 37.5+/-2.5(P<0.05) for the co-localized, respectively. At DIV7 and DIV19, 75.4% and 57.9% NR2B-NMDAR clusters were co-localized with GluR2-AMPAR; and 50.6% and 37.6% GluR2-AMPAR clusters were co-localized with NR2B-NMDAR, respectively.

CONCLUSIONThe density of NR2B-NMDAR containing and GluR2-AMPAR containing receptor clusters increases during development of hippocampal neurons in culture. Although the co-localized clusters are increased as well in an unit length of dendrite, the extent to which the two receptor clusters are co-localized decreases. These data imply a possible change in the partnership of AMPA receptor subtype and NMDA receptor subtype at newly formed synapses during development.

Animals ; Cells, Cultured ; Dendrites ; chemistry ; Hippocampus ; chemistry ; Rats ; Rats, Sprague-Dawley ; Receptors, AMPA ; analysis ; Receptors, N-Methyl-D-Aspartate ; analysis

Objective To explore the effect of multi-disciplinary team (MDT) combined with bundle management on prevention and control of multidrug-resistant organism (MDRO) infection in the intensive care unit(ICU).Methods Patients who were admitted to the ICU in a tertiary first-class hospital from January 2013 to December 2015 were studied,MDT combined with bundle management has been applied in the prevention and control of MDRO infection in ICU since January 2014,continuous quality improvement program was performed one year later,isolation of MDROs from specimens of ICU patients before implementation(in the year of 2013),after implementation(in the year of 2014),and after continuous quality improvement(in the year of 2015) was compared.Results The infection rates of MDROs in ICU patients before implementation,after implementation,and after continuous quality improvement were 26.55％ (154/580),17.13％ (117/683),and 12.01％ (77/641) respectively,showing a downward trend,with a significant difference (x2 =44.030,P＜0.001);the total isolation rates of MDROs in ICU patients were 64.44％(154/239),63.59％(117/184),and 43.26％ (77/178) respectively,showing a downward trend,with a significant difference (x2 =22.284,P＜0.001).The main MDROs in ICU were multidrug-resistant (MDR) and pandrug resistant(PDR) Acinetobacterbaumannii (44.54％).Conclusion MDT combined with bundle management can decrease MDRO infection rate and isolation rate in ICU.

OBJECTIVETo evaluate the trabecular bone micro-structure from different sites of spine in adolescent idiopathic scoliosis patients. The target site consisted of the bilateral facet joints from apical vertebrae and from end vertebrae.

METHODSNine AIS patients with mean age 14.9 years (range 12-17 years) and mean Cobb angle 56 degrees (ranged 48 degrees-84 degrees) were recruited into this study. Corrective surgery was indicated to these patients, and facet joint biopsies were collected during decortications for spinal fusion. Biopsy consents were obtained from patients. Bone specimens were fixed with routine histology procedures and scanned by micro computer tomography (muCT40, Scanco Medical, Switzerland). Ten pairs of facet joint were harvested from apical vertebrae and 12 pairs from end vertebrae. Three-dimensional reconstructed images with the resolution of 20 microm were achieved for histomorphometric analysis.

RESULTSThe values of BV/TV (0.268 vs. 0.354, P < 0.05), TbTh (0.20 vs. 0.24, P < 0.05), TbSP (0.66 vs. 0.56, P < 0.05) and BS/BV (12.7 vs. 10.4, P < 0.05) between convex and concave side at the apex area were significantly different. No difference was found in any structural parameters between left and right side at end area, and upper thoracic (T5, 6) and thoracolumbar (T12, L1).

CONCLUSIONDue to asymmetric compression and tension shared between convex and concave side, more bone and thicker and more profound trabecular bones are observed in the concave side than in the convex side, which seems to resist the progression of spinal curvature. This finding suggests that the provocative factors which cause the progression of the curve in certain patients may not lie in the bone component of spine.

Adolescent ; Child ; Female ; Humans ; Scoliosis ; diagnostic imaging ; pathology ; physiopathology ; Tomography, X-Ray Computed ; Zygapophyseal Joint ; pathology ; physiopathology

BACKGROUNDImmunotherapy is emerging as a promising cure for cancer. However, a severe problem in this area is the immune tolerance to tumor cells and tumor-associated antigens, as evidenced by the ability of cancer to escape immune surveillance. To overcome this problem this work examined the potential of improving the antigenicity of myeloma by metabolic engineering of its cell surface carbohydrate antigens (i.e., glycoengineering) and presentation of the modified tumor antigens by dendritic cells (DCs) to generate cytotoxic T-lymphocytes (CTLs).

METHODSCD138+ myeloma cells were isolated from 11 multipe myeloma (MM) patients by the immunomagnetic bead method. The MM cells were treated with N-propionyl-D-mannosamine (ManNPr), a synthetic analog of N-acetyl-D-mannosamine (ManNAc), the natural biosynthetic precursor of N-acetyl sialic acid (NeuNAc), to express unnatural N-propionylated sialoglycans. The glycoengineered cells were then induced to apoptosis, and the apoptotic products were added to cultured functional DCs that could present the unnatural carbohydrate antigens to autologous T-lymphocytes.

RESULTSIt was found that the resultant DCs could activate CD4+ and CD8+ T-lymphocytes, resulting in increased expression of T cell surface markers, including CD8CD28 and CD4CD29. Moreover, upon stimulation by glycoengineered MM cells, these DC-activated T-lymphocytes could release significantly higher levels of IFN-gamma (P < 0.05). Lactate dehydrogenase (LDH) assays further showed that the stimulated T-lymphocytes were cytotoxic to glycoengineered MM cells.

CONCLUSIONSThis work demonstrated that glycoengineered myeloma cells were highly antigenic and the CTLs induced by the DCs loaded with the unnatural myeloma antigens were specifically cytotoxic to the glycoengineered myeloma. This may provide a new strategy for overcoming the problem of immune tolerance for the development of effective immunotherapies for MM.

Antigens, Tumor-Associated, Carbohydrate ; immunology ; Cytotoxicity, Immunologic ; Dendritic Cells ; immunology ; Humans ; Immunophenotyping ; Immunotherapy ; Interferon-gamma ; biosynthesis ; Multiple Myeloma ; immunology ; pathology ; therapy ; T-Lymphocytes ; immunology

The aim is to observe the expression of human factor VIII gene in mice tranduced in vivo and ex vivo. The vector pLNC-FVIII BD was generated by cloning a B-domain-deleted (760aa-1639aa) FVIII cDNA (FVIIIBD cDNA) into retroviral vector pLNCX. 2 x 10(6) of mouse bone marrow stroma cells transduced by LNC-FVIII BD were infused into 4-week-old BALB/c mice by tail-vein injection. pLNC-FVIII BD was conjugated with PAMAM dendrimer to form complex PAMAM-pLNC-FVIII BD, with which C57BL/6J were injected by tail vein (200 micro l contained 15 micro g/mouse) and sacrificed at days 1, 2, 7, 14, 21 and 28, respectively after injection. Tissue such as liver, spleen, lung and kindney were harvested, with which the transcription were detected by means of RT-PCR. In addition, blood was collected to be measured human FVIII Ag, human FVIIIc and anti-FVIII of human inhibitors. The results showed that the highest level of human FVIII in the recipient BALB/c mice was 8.6 +/- 1.44 ng/ml detected on the first day post-injection; anti-FVIII antibodies were detected from the first week post-injection, and then the level of FVIII Ag decreased and cannot be measured on the fourth week. In the C57BL/6J mice physiological level of human FVIII was expressed in plasma at 48 hours after injection and the average human FVIIIc was 0.62 U/ml and the average human FVIII Ag was 115.5 ng/ml, and gradually reduced later. Anti-FVIII of human inhibitors was not revealed all the time. Syngene image scanning demonstrated that the transcription of the human FVIII BD cDNA occurred mainly in spleen and lung, and secondarily in liver and kidney. No side effects of PAMAM-pLNC-FVIII BD were observed in mice tissue by pathological examination at 4 weeks. In conclusion, retrovirus-transduced bone marrow stroma cells effectively produced human FVIII after ex vivo transduction, but the development of anti-FVIII antibodies in recipient mice influenced the expression level. The human FVIII gene can successfully be transduced in vivo through injecting PAMAM-pLNC-FVIII BD cDNA into mice intravenously. There was physiological level expression of human FVIII in plasma at 48 hours after injection and the average human FVIIIc is 0.62 U/ml and the peak in the six mice was 0.89 U/ml, and gradually reduced later.
Animals ; DNA, Complementary ; analysis ; Factor VIII ; genetics ; Genetic Therapy ; Hemophilia A ; therapy ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Transfection

OBJECTIVETo study the molecular mechanism of antithrombin (AT) gene C2759T (Leu99Phe) mutation causing AT deficiency.

METHODSA mutated AT cDNA expression plasmid ATM2759 was constructed by mega-primer method. ATM2759 and wild type AT cDNA expression plasmid ATN were transfected into COS7 cells or CHO cells by using Superfect reagent respectively for in vitro expression study and immunofluorescence assay.

RESULTSThe antigen levels of AT (AT:Ag) in the cell lysate of ATM2759 transfected COS7 cells and the cell culture supernatant were 174.97% and 35.63% of that of ATN transfected COS7 cells respectively, whereas the AT activity in the cell culture supernatant was 47.73% of the control's. Immunofluorescence analysis showed that the fluorescence intensity was significantly higher in ATM2759 transfected CHO cells than in those transfected with ATN.

CONCLUSIONSLeu99Phe substitution may not affect the binding capacity of AT with heparin. Secretion defect and intracellular accumulation of the mutated AT protein might be the mechanisms of this mutation causing AT deficiency.

Animals ; Antithrombin III ; genetics ; metabolism ; Antithrombin III Deficiency ; genetics ; CHO Cells ; COS Cells ; Cercopithecus aethiops ; Cricetinae ; Cricetulus ; Fluorescent Antibody Technique ; Mutation ; Plasmids ; genetics ; Transfection

OBJECTIVETo investigate the pathogenesis of inherited coagulation factor VII (FVII) deficiency.

METHODSThe diagnosis was validated by coagulant parameter assay. FVII gene mutations were analysed in the proband by DNA direct sequencing of PCR products of all exons, exon-intron boundaries and the 3', 5'untranslated sequences. The mutations were confirmed by reverse sequencing. The ectopic transcripts of RT-PCR were used to confirm the characteristics of the mutation in non-canonical splice site (IVS1a + 5g > a).

RESULTSDouble heterozygous mutations in the propositus were identified: a T to G mutation at position 10961, resulting in His348Gln substitution, a non-canonical splice site (IVS1a + 5g > a) mutation, causing the new model of splice and frameshift mutation.

CONCLUSIONDouble heterozygous mutations of His348Gln and IVS1a + 5g > a were identified in a propositus, the splicing pattern of the IVS1a + 5g > a mutation was reported for the first time.

Amino Acid Sequence ; Base Sequence ; Factor VII ; genetics ; Factor VII Deficiency ; genetics ; Heterozygote ; Humans ; Molecular Sequence Data ; Mutation, Missense ; RNA Splicing