OBJECTIVETo evaluate the biological safety of manufactured heterologous deproteinized bone and to provide an experimental basis for clinical applications.

METHODSDeproteinized bone (10 mm) and leaching liquor were made from pig ribs with a series of physical and chemical methods, then were evaluated through acute and subacute toxicity test, hemolysis test, pyrogen test, intracutaneous test, intramuscular implantation test and cytotoxity test.

RESULTSNo obvious toxicity, hemolysis, pyrogenic characteristics, skin irritation, inflammatory reaction after intramuscular implantation and cytotoxity were observed.

CONCLUSIONSThe heterologous deproteinized bone has good biological safety and meets all the demands of scaffold material for tissue engineering.

Animals ; Bone Transplantation ; Cytotoxicity Tests, Immunologic ; Hemolysis ; Mice ; Rabbits ; Tissue Engineering ; Transplantation, Heterologous

OBJECTIVETo analyze the clinical characteristics of plastic bronchitis associated with 2009 influenza A virus (H1N1) infection.

METHODA retrospective investigation of the clinical manifestation, bronchoscopy, and the histology of the cast, clinical course and outcome of 8 children with plastic bronchitis associated with influenza A virus (H1N1) infection during winter of 2009 and 2010 was performed.

RESULTAll 8 cases were boys, the range of age was 3 to 6 years. Five cases occurred in 2009 winter, accounting for 3.3% (5/150) of hospitalized children with influenza A (H1N1) infection; 3 cases occurred in 2010 winter, accounting for 15.8% (3/19) of hospitalized children with influenza A (H1N1) infection. Two patients had an underlying chronic disease, 1 had asthma, and the other had allergic rhinitis and atopic dermatitis. All the 8 cases had fever, cough and sputum; 2 had wheezing; 5 had respiratory distress. All 8 cases were diagnosed as influenza A virus (H1N1) infection complicated with pneumonia, of whom 5 patients had atelectasis, 2 had pneumothorax, 1 had pneumomediastinum, 1 had parapneumonic effusion, 2 patients were suspected of foreign body aspiration. Seven cases were admitted to an ICU, 5 patients developed respiratory failure, and 3 patients required mechanical ventilation. Flexible bronchoscopy and bronchial lavage was performed in all cases and showed bronchial cast. Histological examination of the bronchial cast revealed a fibrinous material containing large quantity of eosinophils, neutrophils, and lymphocytes in 7 patients, fibrinous material and necrotic material without inflammatory cells in 1 patient. After the bronchial cast was removed, all patients were improved greatly, no patients died.

CONCLUSIONPlastic bronchitis is a life-threatening complication associated with 2009 influenza A (H1N1) virus infection in children. In children with rapid and progressive respiratory distress with lung atelectasis or consolidation on chest radiograph, plastic bronchitis should be considered. Bronchoscopic extraction of casts should be carried out early.

Antiviral Agents ; administration & dosage ; therapeutic use ; Bronchitis ; complications ; diagnosis ; therapy ; virology ; Bronchoscopy ; Child ; Child, Preschool ; Foreign Bodies ; complications ; Glucocorticoids ; administration & dosage ; therapeutic use ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza, Human ; complications ; virology ; Intensive Care Units ; Male ; Pulmonary Atelectasis ; diagnosis ; therapy ; virology ; Rare Diseases ; Respiratory Insufficiency ; diagnosis ; therapy ; virology ; Retrospective Studies ; Tomography, X-Ray Computed ; Treatment Outcome

Objective To investigate the clinical characteristics of plastic bronchitis (PB) so as to improve the awareness of the disease.Methods Twenty-four children with PB were collected from Jul.2009 to Mar.2012 in Shenzhen Children's Hospital.The clinical manifestation,bronchoscopy,histology of the cast,clinical course and outcome were reviewed retrospectively.Results Of the 24 children with PB,18 cases were male,6 cases were female,and the range of age was 1 year and 2 months to 10 years and 3 months,with the median age of 3 years and 4 months.Three patients had an underlying chronic disease,1 case had asthma,1 case had hydronephrosis,and 1 case had ventricular septal defect repair before 1 year and 8 months.All the cases had fever,cough and sputum,while 10 cases had wheeze,and 5 cases had respiratory distress.All cases were diagnosed as pneumonia or severe pneumonia,of which 14 case had atelectasis,10 cases had parapneumonic effusion,5 cases suspected of foreign body inhalation,3 cases had pneumothorax,and 3 cases had mediastinal hernia.Fourteen cases were admitted to PICU,6 patients developed respiratory failure,and 9 patients required mechanical ventilation.Flexible bronchoscopy and bronchial lavage were performed in all cases and showed bronchial cast.Histological examination of the bronchial cast revealed that fibrinous material containing large quantity of eosinophils,neutrophils,and lymphocytes in 23 patients,and no inflammatory cells in 1 patient.After a bronchial cast was removed,all patients were improved greatly,and no patient dead.Conclusions Plastic bronchitis is a rare pediatric critical disease,which has high mortality.In children with rapid and progressive respiratory distress with lung atelectasis,pleural effusion or consolidation on chest radiograph,PB should be considered.Bronchial endoscopy is the most effective method for treatment of PB.

OBJECTIVETo investigate immunity of a recombinant adenovirus vaccine (rAdV) containing codon-modified neuraminidase (Mod. NA) gene of H5N1 influenza virus in BALB/c mice and to screen for appropriate dose.

METHODSBALB/c mice were immunized with the rAdV-Mod.NA vaccine intramuscularly twice (double injection at 0 and 4th week) in three groups, low dosage (10(5) TCID50 per dose), medium dosage (10(7) TCID50 per dose) and high dosage (10(9) TCID50 per dose). The effect of humoral and cell-mediated immunity were analysed at 5th week.

RESULTS(1) The rAdV-Mod.NA vaccine could elicit both humoral and cell-mediated robust NA specific immunity in mice by neuraminidase inhibitor assay and IFN-gamma ELISpot assay; (2) 10(7) TCID50 per dose was the appropriate dose; (3) Peptide NA(109-124): CRTFFLTQGALLNDKH and peptide NA(182-199): AVAVLKYNGIITDTIKSW were the dominant epitopes for neuraminidase-immunized BALB/c mice, which was screened out from the whole length of neuraminidase of an H5N1 virus, A/Anhui/1l/2005.

CONCLUSIONThe recombinant adenovirus NA could induce specific humoral and cellular immune responses in BALB/c after immunization, which suggest rAdV-Mod.NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.

Adenoviridae ; genetics ; metabolism ; Animals ; Dose-Response Relationship, Immunologic ; Female ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; administration & dosage ; genetics ; immunology ; Influenza, Human ; immunology ; prevention & control ; virology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; administration & dosage ; genetics ; immunology ; Random Allocation ; Vaccines, Synthetic ; administration & dosage ; genetics ; immunology ; Viral Proteins ; administration & dosage ; genetics ; immunology

OBJECTIVETo investigate the influence of avian influenza virus (AIV) NS1 protein on the expression of interferon-inducible protein 10 (IP-10).

METHODSNSI gene from virus A/Anhui/1/2005 (H5N1), NS1 gene inserted with 80-84 amino acids from virus A/Anhui/1/2005 (H5N1) and NS1 gene from virus A/Puerto Rico/8/1934 (H1N1) were cloned into the eukaryotic expression vector pEGFP-N1, and transfected into BEAS-2B cells, IP-10 expression level in transfected cells was detected by flow cytometry.

RESULTSCompared with the control group pEGFP-N1, expression of these three different NS1 genes can down-regulate the expression of IP-10 in BEAS-2B cells, but there is no significant difference as to the lower level among them.

CONCLUSIONNS1 protein of A/Anhui/1/2005 (H5N1) can down-regulate the expression level of IP-10, but this may not clarify its relationship with the virulence of AIV.

Cell Line ; Chemokine CXCL10 ; genetics ; metabolism ; Down-Regulation ; Gene Expression ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; metabolism ; Influenza, Human ; genetics ; metabolism ; virology ; Viral Nonstructural Proteins ; genetics ; metabolism

OBJECTIVETo study causes of ineffectiveness of microvascular decompression (MVD) in treatment of hemifacial spasm (HFS).

METHODSReoperative MVD was performed in 23 HFS patients with previous ineffective MVD. In the patients, the main causes of ineffectiveness included misjudgment of compressing vessels (7 patients), improper insertion of decompressing grafts (9), improper selection of grafts (5) and small grafts (2).

RESULTSSymptoms of HFS disappeared immediately after the second MVD in 21 patients and delayed in 2 patients (after 2 weeks, 6 weeks). No recurrence of HFS was noted during the follow-up period of 1.0 - 6.0 years (mean 3.4 years).

CONCLUSIONSMVD is an effective microsurgical method for treating HFS. Accurate judgement of compressing vessels and proper decompression are the key to surgical effectiveness.

Adult ; Decompression, Surgical ; methods ; Female ; Follow-Up Studies ; Hemifacial Spasm ; surgery ; Humans ; Male ; Medical Futility ; Microsurgery ; methods ; Middle Aged ; Reoperation ; Treatment Outcome ; Vascular Surgical Procedures ; methods ; Young Adult

OBJECTIVETo observe the effect of tetrandrine on activity of collagenase derived from human hypertrophic scar for the sake of clarifying the mechanism as tetrandrine acting on scar.

METHODSThe experimental concentration was controlled below that of cell proliferation inhibited, SDS-PAGE electrophoresis was adopted to separate collagenase from extracellular matrix, and then activated by trypsin analyzed the activity of collagenase with density scanning apparatus. At the same time quantity of extracellular collagen was measured using improved chloraseptine T oxidizing assay, moreover analyzed correlation between activity of collagenase and quantity of extracellular collagen.

RESULTSIn the concentration below the lever of inhibiting fibroblast proliferation, the total activity of collagenase could be significantly increased by tetrandrine with dosage-dependence associated with quantity of extracellular collagen reduced, which was much greater than that of triamcinolone.

CONCLUSIONIncreasing activity of collagenase on degradation of collagen even in a lower concentration was one of the mechanisms of tetrandrine treating hypertrophic scar.

Benzylisoquinolines ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Collagenases ; metabolism ; Fibroblasts ; cytology ; Humans

The aim of this study is to develop the recombinant adenovirus vaccine (rAdV) candidates containing neuraminidase (NA) gene of H5N1 influenza virus and test in BALB/c mice the effect of cell-mediated immunity. In this study, two kind of NA gene (WtNA gene, the wild type; Mod. NA gene, the codon-modified type) derived from H5N1 influenza virus (A/Anhui/1/2005) were cloned and inserted respectively into plasmid of adenovirus vector, then the rAdV vaccines candidates (rAdV-WtNA and rAdV-Mod. NA) were developed and purified, followed by immunization intramuscularly (10(9) TCID50 per dose, double injection at 0 and 4th week) in BALB/c mice, the effect of cell-mediated immunity were analysed at 5th week. Results indicated that: (i) NA protein expression was detected in two rAdV vaccines candidates by Western blotting; (ii) the rAdV-Mod. NA vaccine could elicit more robust NA specific cell-mediated immunity in mice than that of rAdV-WtNA vaccine (P = 0. 016) by IFN-gamma ELIspot assay. These findings suggested rAdV-Mod. NA vaccine was a potential vaccine candidate against H5N1 influenza and worthy of further investigation.
Animals ; Blotting, Western ; Female ; Humans ; Immunity, Cellular ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Influenza Vaccines ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Neuraminidase ; genetics ; immunology ; Orthomyxoviridae Infections ; immunology ; virology ; Polymerase Chain Reaction ; Random Allocation ; Viral Proteins ; genetics ; immunology

In order to improve the expression of human avian influenza virus hemagglutinin (HA) and meet the pandemic influenza vaccine needs, we optimized and synthesized the whole length of HA gene of H5N1 (A/Anhui/1/2005) influenza virus in accordance with the human's codon preference, and inserted it to the eukaryotic expression vector pDC315 to construct a eukaryotic expression vector pDC315-Mod. HA. This plasmid and the eukaryotic expression vector pDC315-Wt. HA containing wild HA gene were transfected into 293T cells respectively to compare the expression of HA protein. The results showed that according to the comparison and identification by indirect immunofluorescence assay and Western blot test, the expression of HA protein in 293T cells was significantly improved after codon optimization. This laid a foundation for pandemic influenza vaccine research.
Blotting, Western ; Cell Line ; Codon ; Fluorescent Antibody Technique, Indirect ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H5N1 Subtype ; genetics ; Plasmids

OBJECTIVETo investigate the inductive therapeutic effects of imatinib combined with VP low dose regiment on adult patients with Ph-positive acute lymphoblastic leukemia (Ph(+) ALL).

METHODSFourteen newly diagnosed adult patients with Ph(+) ALL were treated with VP regimen, and imatinib (400 mg/d) was added at the 8(th) day. VP regimen would be stopped when neutropenia lasted for 1 week or complicated with infection, fever, etc. Therapeutic effects were assessed by bone marrow morphology and quantitative analysis of BCR/ABL:ABL at the 28(th) - 33(rd) day. Patients could be treated with imatinib combined with chemotherapy for consolidation and maintenance therapy or were treated with allogeneic hematopoietic stem cell transplantation after complete remission.

RESULTSFourteen cases obtained CR1 after first course of treatment, the median decline of BCR/ABA:ABL was 55.89 (10.25 -180.97) %; during the induction chemotherapy, pulmonary infection occurred in 3 patients, diarrhea in 1 patients, facial edema in 3 patients, however, all these patients were cured after symptomatic treatment, only 1 patient died of relapse after transplantation.

CONCLUSIONIn the period of tyrosine kinase inhibitor (TKI), inductive chemotherapy combined with imatinib and low dose VP can obtaine satisfactory CR rate and decrease the toxicity of the traditional drugs. It is suggested that TKI combined with VP regimen chemotherapy can achieve CR1 and make possible for allo-HSCT, from which patients can achieve the long-term survival.

Adult ; Antineoplastic Combined Chemotherapy Protocols ; Bone Marrow ; Cisplatin ; Fusion Proteins, bcr-abl ; Hematopoietic Stem Cell Transplantation ; Humans ; Imatinib Mesylate ; Induction Chemotherapy ; Neutropenia ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Protein Kinase Inhibitors ; Recurrence ; Remission Induction ; Transplantation, Homologous ; Vindesine