OBJECTIVETo compare two different methods, namely orthotopic implantation and armpit implantation, for establishing nude mouse model bearing osteosarcoma in light of the tumor formation rate and time, tumor growth changes and lung metastasis.

METHODSOsteosarcoma cells were inoculated in the left armpit or the femoral medullary canal of nude mice, and the tumor growth was observed 6 weeks later. The differences in the pulmonary metastasis were examined macroscopically and microscopically.

RESULTSArmpit implantation allowed easy operation and fast tumor growth, but tumors often sustained damages by frictions on the surface (P<0.05), and avascular necrosis occurred earlier in the tumors (P<0.05). With this approach, the pulmonary metastasis rate was only 40% (P<0.05), and the metastases in the lungs were sparser (P<0.05) and small. Orthotopic transplantation could well simulate the original growth environment of osteosarcoma, and resulted in milder avascular necrosis and greater pulmonary metastasis rate (100%) with larger and denser metastatic foci. But orthotopic transplantation required more complicated operation for establishing osteosarcoma-bearing models.

CONCLUSIONThe two methods both result in high tumor growth rate, but orthotopic transplantation is superior in inducing pulmonary metastasis of osteosarcoma and reducing friction injury of the tumor and avascular necrosis in the tumor.

Animals ; Bone Neoplasms ; Disease Models, Animal ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; methods ; Osteosarcoma

Objective：This study was designed to establish and optimize the research methods for proteome,and to analyze the proteome components of human lung squamous carcinoma cell line NCI H520. Methods： A series of methods, including immobilized pH gradient two dimensional polyacrylamide gel electrophoresis(2DE), silver staining, PDQuest 2DE analysis software, peptide mass fingerprint based on matrix assisted laser desorption/ionization time of flying mass spectrometry (MALDI TOF MS) and SWISS PROT database searching, were used to separate and indentify the proteome of human lung squarmous carcinoma cell line NCI H520. Results： The good 2DE pattern including resolution and reproducibility was obtained. After silver staining, the 2DE image analysis by PDQuest 2DE software had detected average of 1146± 116 spots, and 851± 95 spots were matched. The average matching rate was 73.3％ . There had a good reproducibility of spot position in 2DE map, with average deviation in IEF direction of 1.52± 0.22 mm， while in SDS PAGE direction it was 1.97± 0.13 mm. Sixty spots were incised from silver staining gel randomly and digested in gel by TPCKtrypsin. Fifty four peptide mass fingerprints (PMF) maps were obtained by MALDI TOF MS. The typic peptide masses were searched in the SWISS PROT database by PeptIdent software. Forty four proteins were preliminarily identified. Some of them were cell cycle related proteins such as Cyclin H, some were signal transduction related proteins such as mitogen activated protein kinase, protein kinase C and receptor protein tyrosine kinase ERBB 2, some were oncogene related proteins such as Ras related protein RAB 36, etc. Conclusions： The main proteome research system including IPG 2DE, image analysis, MALDI TOF MS derived PMFs and database searching has been established. The data of NCI H520 obtained by above methods will be useful for the establishment of human lung squamous cell proteome database.

Objective To investigate the prognostic value of acute beart block (AHB) after percutaneous transluminal septal myocardial ablation (PTSMA) in patients with hypertrophic obstructive cardiomyopathy (HOCM). Methods Ninety-four HOCM patients underwent PTSMA were included in this study. Twelve-lead electrocardiograms were obtained during and post PTSMA. Association between clinic events and incidence of post-PTSMA AHB was analyzed. Results AHB was induced in 26 patients by PTSMA and disappeared in 11 patients shortly post PTSMA, subacute intraventricular conduction disturbances was seen in 11 (42.3%), subacute I°AVB in 2(7.7% ) and subacute Ⅲ°AVB in another 2 (7.7%) patients. Among 68 patients without AHB during PTSMA, intraventricular conduction disturbances was evidenced in 14 patients (20.6% ), I°AVB in 2 (2.9% ) and Ⅲ°AVB in 1 patient (1.5%) after PTSMA. AHB patients with subacute heart block were associated with poor prognosis (conduction block duration was 42.00 h) while patients without AHB was associated with benign prognosis even with new onset of subacute heart block (conduction block duration was 7.33 h, P <0.01). Conclusion Patients with AHB during PTSMA are at higher risk for subacute heart block, especially intraventricolar conduction disturbances. AHB patients with subacute heart block were associated with poor prognosis and longer recovery time conducting system.

The aim of this study was to explore application value of detecting platelet associated antibody and platelet membrane glycoprotein in the diagnosis and prognosis for immune thrombocytopenia. The platelet associated immunoglobulin (PAIg) and platelet membrane glycoprotein (CD41, CD61, GPIIb/IIIa) in 76 cases of immune thrombocytopenia and 30 healthy subjects were determined by FCM. The results showed that PAIg level in ITP patients included PAIgG (31.25 +/- 18.06)%, PAIgM (32.41 +/- 15.51)%, PAIgA (23.39 +/- 16.67)% which were remarkedly higher than in health control (10.48 +/- 5.05)%, (9.40 +/- 4.42)% and (7.23 +/- 3.61)% (P < 0.001). In patients with secondary immune thrombocytopenia (chronic aplastic anemia, SLE, Evans syndrome, liver cirrhosis hypersplenism, etc), PAIg level was higher than that in control group, while the platelet membrane glycoprotein in the blood of these patients was lower than that in control group. The level of PAIg decreased (P < 0.05) after treatment, but platelet membrane glycoprotein increased (P < 0.01). The result suggested that measurements for platelet membrane glycoprotein and platelet associated antibody by FCM were practical with high sensitivity, rapidity and simplicity used as a routine method in diagnosis and evaluation of the therapeutic effects in immune thrombocytopenia patients.
Adolescent ; Adult ; Aged ; Blood Platelets ; immunology ; Child ; Female ; Flow Cytometry ; Humans ; Immunoglobulins ; blood ; Integrin beta3 ; analysis ; Male ; Middle Aged ; Platelet Glycoprotein GPIIb-IIIa Complex ; analysis ; Platelet Membrane Glycoprotein IIb ; analysis ; Platelet Membrane Glycoproteins ; analysis ; Purpura, Thrombocytopenic, Idiopathic ; blood ; diagnosis ; Thrombocytopenia ; blood ; diagnosis

OBJECTIVETo elucidate the effects of interferon-gamma (IFN-gamma) on the transforming growth factor beta (TGF-beta)/Smad pathway in keloid-derived fibroblasts (KFb), and to investigate the underlying mechanism in the treatment of pathologic scar with IFN-gamma.

METHODSKeloid tissue of 3 patients were obtained, and then KFb were separated and cultured in vitro. KFb from passages 3 to 5 were used for the study. (1) KFb were divided into control group (incubated with serum-free DMEM), TGF-beta(1) group (treated with 10 ng/mL TGF-beta(1)), IFN-gamma group (treated with 100 ng/mL IFN-gamma), and TGF-beta(1)+IFN-gamma group (incubated with 10 ng/mL TGF-beta(1) combined with 100 ng/mL IFN-gamma). The expression level of mRNA and protein of connective tissue growth factor (CTGF), alpha smooth muscle actin (alpha-SMA) protein and expression of alpha-SMA positive KFb were detected by real-time fluorescent quantitation RT-PCR (FQ-RT-PCR), Western blot and immunofluorescence cytochemical staining. (2) Another sample of KFb was obtained and treated with 10 ng/mL IFN-gamma. The expression level of Smad 3 and Smad 7 protein was detected by Western blot before and 1, 2, 4, 6, 8 h post stimulation (PSH). The expression level of Smad 3 and Smad 7 mRNA was assessed by FQ-RT-PCR before stimulation and 30 mins post stimulation and at PSH, 1, 2, 4, 6, 8. (3) Another sample of KFb was obtained and divided into 1, 10 and 100 ng/mL IFN-gamma groups based on the concentration of IFN-gamma, treated for 4 hours; KFb without IFN-gamma treatment was set up as control group. The expression levels of the protein and mRNA of Smad 3 and Smad 7 were measured by FQ-RT-PCR and Western blot.

RESULTS(1) The level of mRNA and protein of CTGF in IFN-gamma group (0.017 +/- 0.009 and 1.198 +/- 0.004) was respectively lower than that in control group (0.024 +/- 0.013 and 1.229 +/- 0.011, P < 0.05). The level of mRNA and protein of CTGF in TGF-beta(1)+IFN-gamma group (0.634 +/- 0.138 and 1.204 +/- 0.010) was respectively lower than that in TGF-beta(1) group (1.331 +/- 0.298 and 1.727 +/- 0.004, P < 0.01). The fluorescence intensity of alpha-SMA positive KFb (0.922 +/- 0.059) and the expression level of alpha-SMA protein (0.3051 +/- 0.0031) in IFN-gamma group decreased significantly than those in control group (1.055 +/- 0.005 and 0.4513 +/- 0.0094, P < 0.01). The fluorescence intensity of alpha-SMA positive KFb (1.129 +/- 0.004) and the expression level of alpha-SMA protein (0.6734 +/- 0.0098) in TGF-beta(1)+IFN-gamma group decreased significantly than those in TGF-beta(1) group (1.270 +/- 0.005 and 1.3842 +/- 0.0024, P < 0.01). (2) The expression level of Smad 3 mRNA and protein at the first time point after IFN-gamma treatment increased temporarily then decreased gradually, and mRNA expression level reached the nadir at PSH 4, it rose gradually later, though it was still lower at PSH 8 than that before treatment (P < 0.01); protein expression level at PSH 8 was significantly lower than that before treatment (P < 0.01). The expression level of Smad 7 mRNA and protein increased gradually to the maximum at PSH 2 and 4 respectively, then decreased but was still higher at PSH 8 than that before treatment (P < 0.05). (3) Compared with those in control group, the expression levels of Smad 3 mRNA and protein in 1, 10 and 100 ng/mL IFN-gamma group were significantly lower, the expression levels of Smad 7 mRNA and protein were significantly higher (P < 0.05 or P < 0.01). The higher concentration of IFN-gamma, the more significant differences were observed.

CONCLUSIONSIFN-gamma can down-regulate the expression of Smad 3 while up-regulate the expression of Smad 7 in a time- and dose-dependent manner, and reduce the expression level of CTGF and alpha-SMA in the basic state or induced by TGF-beta(1), which shows a significant inhibitory effect on the TGF-beta/Smad signal pathway. This may be an important mechanism in the treatment of pathologic scar by IFN-gamma.

Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Humans ; Interferon-gamma ; pharmacology ; Keloid ; metabolism ; RNA, Messenger ; genetics ; Signal Transduction ; drug effects ; Smad Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism

Objective To assess the therapeutic effect of gamma knife radiosurgery for residual tumors after surgical resection of hemangioblastoma in children. Methods The clinical data were retrospectively analyzed in 9 children who received gamma knife radiosurgery for the residual tumors after a previous surgery for hemangioblastoma resection. Results The 9 children were followed up for 12-48 months, and 6 children showed a tumor volume reduction by over 50% with obviously improved clinical symptoms. One child showed no response to the treatment and received a second gamma knife radiosurgery. In 2 children, the tumor volume increased progressively after the radiosurgery, and a second radiosurgery was performed in 1 case and open surgery in the other. Conclusion Gamma knife radiosurgery is safe and effective for treatment of the residual tumors in children with previous surgical resection of hemangioblastoma.

Objective To identify the serotype of 73 non-polio enterovirus (NPEV) strains from acute flaccid paralysis (AFP) cases in Ningxia province,during 1997-2011.Methods Partial sequencing of the VP1 region was amplified by RT-PCR with degenerate primers and sequenced while sequences were compared with the database of GenBank by the BLAST algorithm.Evolution was analyzed by constructing phylogenetic tree using Mega 5.1.Results In this study,a total of 73 NPEVs were analyzed,including 4 strains un-typed,69 strains typed by RT-PCR.A total of 27 serotypes were identified,including 8 serotypes of human enterovirus (HEV)-A,19 serotypes of HEV-B.The HEV-B group (46/69,66.7％) constituted the largest proportion of isolates,followed by HEV-A (23/69,33.3％),but no strains were found that belonged to HEV-C or HEV-D group.In the 69 strains,enterovirus 71 was the most frequently seen isolates,followed by coxsackie-virus A4,16,9 and echovirus 24,6.Conclusion HEV-B was the most predominant (46/69,66.7％) serotype of NPEV in Ningxia during the AFP surveillance,in 1997-2011.

Philadelphia chromosome-positive acute myeloid leukemia is controversial and difficult to distinguish from the blast phase of chronic myeloid leukemia.As a myeloid neoplasm,rare cases of this leukemia manifest multiple soft-tissue tumors or bone lytic lesions.In this paper,we describe a 49-year-old male patient who had an abrupt onset with sharp chest pain,fever,fatigue,emaciation,and splenomegaly.18F-fluoro-deoxy-glucose positron emission tomography/computed tomography (18F-FDG PET/CT) result showed diffuse and uneven hypermetabolic lesions in the bone marrow with peripheral bone marrow expansion,multiple soft tissue neoplasms with high 18F-FDG uptake,and lytic bone lesions.Bone marrow smear and biopsy detected aberrant blast cells expressing myeloid rather than lymphoid immunophenotype marker.For the existence of Philadelphia chromosome and BCR-ABL1 fusion gene together with complex chromosome abnormalities,a diagnosis of Philadelphia-positive acute myeloid leukemia was made,although the type (de novo or blast crisis) remained unclear.

OBJECTIVEThe aim of this study is to analyse the efficacy and toxicity of CEOP regimen in the treatment of non-Hodgkin's lymphoma (NHL).

METHODSFrom January 1995 to December 2000, 121 patients with NHL were treated by CEOP regimen with or without radiotherapy for the involved field. The clinical characteristics, response, toxicity and long-term survival results were analysed retrospectively.

RESULTSOf these 121 patients, 83 (68.6%) had B-cell NHL and 38(31.4%) peripheral T or NK-cell NHL; 55. 4% (67/121) had early disease (stage I or II), and 89.3% (108/121) had IPI score 0-2. The median age was 53 years (range: 7-79 yr). All patients were treated by CEOP regimen (totally, 471 cycles) with or without radiotherapy. The overall response (OR) rate in this series was 90.9% (110/121) with a complete remission (CR) rate of 71.9% (87/121); whereas the response rate of chemotherapy alone was 88.4% (107/121) with a CR rate of 67.8% (82/121). Major toxicity consisted of grade III-IV myelosuppression (11.9%), neutropenia (1.9%) and thrombocytopenia and anemia (1.1%). Alopecia was observed in 46.3%. However, cardiotoxicity was mild and reversible. Median follow-up duration in this series was 63 months (range: 2-116 months). The overall 1-, 3- and 5-year survival rate was 84.8%, 62.7% and 55.9%, respectively, with a median survival time of 85 months (2-118 months).

CONCLUSIONOur data show that CEOP regimen combined with or without radiotherapy for the involved field is effective and well tolerated by the patients with non-Hodgkin's lymphoma.

Adolescent ; Adult ; Aged ; Alopecia ; chemically induced ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Child ; Combined Modality Therapy ; Cyclophosphamide ; adverse effects ; therapeutic use ; Epirubicin ; adverse effects ; therapeutic use ; Female ; Follow-Up Studies ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; radiotherapy ; Lymphoma, Non-Hodgkin ; drug therapy ; pathology ; radiotherapy ; Lymphoma, T-Cell ; drug therapy ; pathology ; radiotherapy ; Male ; Middle Aged ; Neoplasm Staging ; Neutropenia ; chemically induced ; Prednisone ; adverse effects ; therapeutic use ; Remission Induction ; Retrospective Studies ; Survival Analysis ; Thrombocytopenia ; chemically induced ; Vincristine ; adverse effects ; therapeutic use

Objective To identify the epidemic characteristics and risk factors of an emerging infectious disease-severe fever with thrombocytopenia syndrome (SFTS) in Hubei province.Methods Active surveillance program on SFTS was set up in monitoring sites-hospitals,at the township level or above,in Suizhou,Huanggang and Wuhan from January to December,2010.Specific surveillance program on SFTS was launched across the province in hospitals above the county level.Cases that matched the definition of surveillance case were identified and reported to Centers for Disease Control and Prevention (CDCs).Cases were interviewed and their blood samples collected and detected using PCR and virus isolation.We also conducted serum antibody surveys among healthy population and livestock and surveillance on vector ticks in those high-epidemic areas.Results 188 cases that matched the definition of surveillance case and 21 deaths were reported in 11 cities,32 countries and 100 towns in 2010,with an incidence rate of 0.33/106.The fatality rate was 11.2％.Data showed that the patients were from hilly areas at the altitude elevated between 28-940 meters.The epidemic period was between April and December with the peak from May to September.The youngest case was an 11-year old,while the eldest was 81 with median age as 56-year old.95.3 ％ of the patients were farmers.All Patients did not have the history of traveling,two weeks before the onset of SFTS.93.6％ of the patients engaged in different kind of work which was associated with agriculture.52.8％ of the patients had been exposed to ticks.22.0％ of the patients had been bitten by ticks.Skin injury was found in 64.2％ of the patients.Samples from 129 cases (68.6％) were collected and detected,with 67.4％ of them (87 cases) showed positive by Real time-PCR for SFTS virus.An elevation in antibody titer by a factor of four or evidence of sero-conversion was observed in 11 patients; SFTS virus was isolated from 2 patients.The total antibody positive rates were 3.8％,55.0％ (6/11 ),36.7％ (2/3) and 80.0％ (4/5) respectively in healthy population,dogs,sheep and cows.Ticks from grass,cattle and sheep were detected positive by Real time-PCR.Conclusion Most cases of SFTS in Hubei were infected by SFTS virus,and cases of livestock were infected by SFTS virus.Ticks might serve as an important vector.Skin injury,exposure to tick bites seemed to be the risk factors.