Aged, 80 and over ; Heart Valve Prosthesis Implantation ; methods ; Humans ; Male ; Prosthesis Failure

OBJECTIVE: To establish the model of skin scald in mice for the study of skin thermal injuries. METHODS: After anaesthetization mice were scalded in a 1 cm-diameter circle area on the central dorsum by boiling water at contact times: 10s or 25s. The mice were sacrificed at 1, 3, 5, 7 and 10 days after scald. The skin samples were collected and analyzed by gross and histopathological examinations. RESULTS: Deep II degree thermal injury involving full-thickness skin was observed in the 10s scald group. III degree thermal injury involving full-thickness skin and the dorsal skeletal muscle was observed in the 25 s scald group. CONCLUSION A mouse skin scald model is established which is stable and can be used on the skin thermal injury in future research.
Animals ; Burns/pathology* ; Disease Models, Animal ; Epithelium/pathology* ; Female ; Hot Temperature/adverse effects* ; Male ; Mice ; Random Allocation ; Reproducibility of Results ; Severity of Illness Index ; Skin/pathology* ; Time Factors

Objective To evaluate the safety and efficacy and summarize the initial experience of transcatheter aortic valve implantation (TAVI) for treating patients with severe aortic stenosis.Methods From October 2010 to May 2011,TAVI using 18 F Corevalve system was applied in 3 patients with severe calcified aortic valve stenosis at high risk for surgery.The efficacy and complications of the procedure were analyzed and the procedure experiences were summarized.Results TAVI procedure was successful in all 3cases.The mean operation time was ( 109.0 ± 22.6) minutes and X-ray exposure time was (24.0 ± 9.5 )minutes.The peak pressure gradients after surgery were significantly reduced [ from ( 84 ± 15 ) mm Hg (1 mm Hg=0.133 kPa) to (6 ± 3) mm Hg].A trivial to mild paravalvular leak was observed in all patients post procedure.Case 1 was free from perioperative complications.Case 2 experienced a transient complete left bundle branch block.Case 3 developed 3 degree atrioventricular block and implanted with a permanent cardiac pacemaker,cardiac tamponade which was relieved through conservative treatment,including pericardial puncture and drainage and acute kidney injury.Conclusions Our initial experience showed that TAVI using the 18 F Corevalve system is safe and effective for patients with severe calcified aortic valve stenosis at high-risk for surgery,though the procedure may cause some complications.Strict patient selection and proficient surgical techniques may reduce the incidence of complications.

OBJECTIVETo develop a novel dual-modified vaccine, the superantigen-linked intestine-carcinoma cells expressing membrane-bound heat shock protein 70 (HSP70), and further examine its anticancer therapeutic effect.

METHODSThe pre-established intestine carcinoma CT26 line expressing membrane-bound heat shock protein 70 (HSP70) was amplified and incubated with superantigen fusion protein, staphylococcal enterotoxin A (SEA) fused with transmembrane sequence (SEA-TM), thereby the dual-modified vaccine was prepared after inactivation. The anticancer efficacy of the vaccine was examined.

RESULTSThe laser confocal microscopy and flow cytometry showed that there co-existed much HSP70 and SEA on the vaccine membrane surface. Both of the single-modified vaccines, the SEA-linked vaccine and membrane-bound-HSP70-expressing one, displayed marked tumor suppression, a prolonged survival period, augmented lymphocyte proliferation and higher NK and CTL activity in the vaccinated mice when compared with its counterpart. Furthermore, the dually modified vaccine induced lymphocyte proliferation most intensively, generated the highest NK and CTL activity as well as the strongest tumor rejection in the vaccinated mice. The survival period of the mice was further prolonged.

CONCLUSIONA new vaccine, SEA-linked and membrane-bound-HSP70-expressing intestine-carcinoma cells can induce more potent anticancer immunity and produce better therapeutic efficacy.

Animals ; Cancer Vaccines ; therapeutic use ; Cell Line, Tumor ; Cell Membrane ; metabolism ; Enterotoxins ; immunology ; Gene Expression ; Genetic Vectors ; HSP70 Heat-Shock Proteins ; genetics ; immunology ; Mice ; Mice, Inbred BALB C ; Superantigens ; immunology ; Transfection

OBJECTIVETo clarify the diagnosis of one suspected case of diphtheria in Guangdong province by epidemiological analysis and etiologic detection.

METHODSOn July 6th 2010, the corynebacterium diphtheria was detected from the nasal secretions of one nasopharyngeal carcinoma patient in a college-town hospital in Guangzhou City, Guangdong Province. The patient and the close contacts were asked to participate in the epidemiological survey; and their nasopharyngeal swabs (3 samples) and the nasal secretions of the patient (1 sample) were collected. The bacteria of the samples were isolated and cultured by blood plate and agar loefflera. The smears of positive strains were dyed and identified by BioMerieux API Coryne biochemical card. Gene tox of β-Corynebacteriophage, Corynebacterium diphtheriae was tested by PCR method, the aliphatic acid was analyzed by gas chromatography method and the Corynebacterium diphtheriae (CMCC 38009) was selected as positive control.

RESULTSThe patient had not gone out, neither had been visited. The patient denied history of vaccines or the immunizations. From the survey on patient's family members and close contacts, no similar symptoms had been found. One strain of Corynebacterium diphtheriae was isolated from the patient's nasal secretions, Gram positive and shape diversified. After cultured by agar loefflera and Gram-dyed and Neisser-dyed, one end or both two ends of the strain showed typical metachromatic granule. API Coryne was identified to Corynebacterium diphtheriae mitis/belfanti (99.4%). The result of gas chromatography method also indicated Corynebacterium diphtheriae. No Corynebacterium diphtheriae was isolated from the nasopharyngeal swabs, neither of the patient nor of the close contacts. The gene tox of β-Corynebacteriophage, Corynebacterium diphtheriae was negative according to the PCR test.

CONCLUSIONThe isolated Corynebacterium diphtheriae did not produce toxin as there was no biological structure gene of toxin. The patient was a health carrier of nontoxic Corynebacterium diphtheriae.

China ; epidemiology ; Corynebacterium diphtheriae ; isolation & purification ; Diphtheria ; epidemiology ; microbiology ; Female ; Humans ; Middle Aged ; Nasopharynx ; microbiology ; Polymerase Chain Reaction ; methods

Objective To evaluate the long-term efficacy of percutaneous pulmonary valve implantation (PPVI) and the durability of the home-made self-expanding pulmonary valve (Venus-P).Methods From May,2013 to Nov.,2015,14 patients who underwent percutaneous pulmonary valve implantation at Zhongshan Hospital,Fudan University and received at least 1 year follow-up were enrolled,including 3 males and 11 females,with an average age of (35.8 ± 7.8) years.All patients with tetralogy of Fallot received radical resection and developed severe pulmonary regurgitation.The longterm mortality,the operation related complications,the short term and long-term effect of PPVI,as well as the durability and effect of the self-expanding pulmonary valve were evaluated in the 14 patients.Results Over an average follow-up period of (2.3 ± 0.8) years (1.0-3.5 years),only 1 patient died (6.7 ％).During the follow-up,no deterioration,infective endocarditis,malignant arrhythmia and other serious complications was observed,and nobody needed reoperation.There was no valve displacement,valve stent fracture,obvious circumferential leakage and pulmonary regurgitation.After PPVI,an acute improvement in pulmonary artery diastolic pressure was observed [(4.93 ± 3.37) mmHg vs.(11.47 ± 4.61) mmHg,P＜0.05].Six month postoperatively,right ventricular end diastolic volume measured by cardiac nuclear magnetic resonance was significantly reduced [(139.29± 18.21)mL/m2 vs.(83.03 ± 20.0) mL/m2,P＜0.05].At 1 year follow up,the across valve pressure difference were (20.85 ± 4.45) mmHg calculated by the echocardiography,and the NYHA cardiac function (Ⅰ-Ⅲ:4 cases;Ⅰ-Ⅱ:10 cases) was improved 1-2 degree and the distance of 6-minute walk test were significantly increased [(475.00 ± 55.06) m vs.(594.23 ± 194.51) m,P＜0.05].Meanwhile,the QRS duration decreased was also observed.The changes of the QRS duration have statistical significance after 1 and 3 months of the PPVI when compared with the baseline [(169.93 ± 21.34) ms vs.(159.87 ± 24.4) ms or (160.00 ± 27.0 ms,P＜0.05].Conclusions PPVI using home-made self-expanding pulmonary valve (Venus-P) for chronic pulmonary regurgitation has good long-term efficacy and low complication rate,and the valve is durable.

OBJECTIVE: To investigate the time-dependent recruitment and differentiation of fibrocytes in skin wound healing. METHODS: Fibrocytes (expressing CD45 and procollagen I ) and myofibroblasts (expressing CD45 and alpha-SMA) were co-localized by immunofluorescent staining. The number of fibrocytes and myofibroblasts was counted at different post-wounding interval. RESULTS: At 3 d after injury, fibrocytes started to recruit at the margin of the wounds. At 5 d after injury, myofibroblasts started to appear in new formed granulation tissue. The number of fibrocytes and myofibroblasts peaked at 7 d post-wounding. CONCLUSION During skin wound healing, myofibroblasts in granulation tissue originated at least partly from fibrocytic differentiation. The time-dependent recruitment and differentiation of fibrocytes may provide new information for wound age determination.
Animals ; Cell Count ; Cell Differentiation ; Disease Models, Animal ; Fibroblasts/metabolism* ; Leukocyte Common Antigens/metabolism* ; Male ; Mice ; Mice, Inbred BALB C ; Myofibroblasts/metabolism* ; Skin/pathology* ; Staining and Labeling ; Time Factors ; Wound Healing

OBJECTION: To investigate the time-dependent appearance of circulating fibrocytes of skeletal muscle in rats after contusion. METHODS: The model of skeletal muscle wound was established in rat. The circulating fibrocytes in contused skeletal muscle were detected by CD45 and procollagen I double immunofluorescence staining method. RESULTS: In the control group, CD45- and procollagen I-positive cells were not detected in skeletal muscle. A few CD45 cells were observed aged from 6 h to 1 d after contusion. A few CD45- and procollagen I-positive cells (fibrocytes) initially gathered in injury area 3d after injury. The ratio of positive fibrocytes significantly increased 5 d after injury. The ratio of fibrocytes was highest at 7 d after contusion and then decreased. The volume of fibrocytes showed bigger with injury time increase compared with 3 d group. The expression of procollagen I and CD45 were weakened at 14d after injury. CONCLUSION The circulating fibrocytes are detected in contused skeletal muscle in time-dependent pattern. Circulating fibrocytes may be a marker in the wound age determination for contused skeletal muscle.
Animals ; Biomarkers/metabolism* ; Collagen Type I/metabolism* ; Contusions/pathology* ; Disease Models, Animal ; Forensic Pathology ; Immunohistochemistry ; Leukocyte Common Antigens/metabolism* ; Male ; Mesenchymal Stem Cells/metabolism* ; Microscopy, Fluorescence ; Muscle, Skeletal/pathology* ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors ; Wound Healing

To develop a stable cell line that could express the RSV NS1, the full-length RSV NS1 gene was generated by RT-PCR amplification from respiratory syncytial virus. NS1 gene was ligated with pBABE-puro to construct the recombinant retroviral expression plasmid pBABE-NS1, which was cotransfected into 293FT packaging cells with PIK packaging plasmid by calcium phosphate co-precipitation. The supernatant of 293FT was collected to infect HEp-2 cells, the resulting cell clones stably expressing NS1 were screened by puromycin. Using QPCR, CPE staining method and indirect immunofluorescence assay, the expression of NS1 at both gene and protein levels was identified. The recombinant plasmid pBABE-NS1 was identified by EcoRI and BamHI endonuclease digestion and the sequence analysis. QPCR results showed that the NS1 gene amplification in HEp-2-NS1 cells was 8483 fold higher than that in HEp-2 cells. Although the exogenous interferon was added, all cells were destroyed after 48 hours post infection using CPE staining method, showing that HEp-2-NS1 cells remained sensitive to the VSV virus. The results of RT-PCR and indirect immunofluorescence assay showed that the NS1 gene in HEp-2 cells could not only transcribe mRNA, but also express NS1 protein steadily. We had successfully established HEp-2-NS1 cell lines with stable expression of respiratory syncytial virus non-structural protein NS1.
Cell Line, Transformed ; HEK293 Cells ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Respiratory Syncytial Viruses ; genetics ; Viral Nonstructural Proteins ; biosynthesis ; genetics