Female ; Hemophilia A ; complications ; Humans ; Middle Aged ; Myasthenia Gravis ; complications

No abstract available.
Drug Therapy* ; Lymphoma*

Objective To study the biological characteristics of CD+44 stem cells in human laryngeal carcinoma.Methods Tumor samples were obtained from 5 patients, and then the human laryngeal carcinoma cells were cultured in vitro by primary tissue culture technique.Taking CD44 molecule as a marker to isolate CD+44 subpopulation cells from laryngeal carcinoma cells for further study.CD+44 and CD-44 cells were cultured and observed fex their development.CD+44 and CD-44 cells were compared in their functional status ( mRNA), cell cycles (GO/G1), their degree of differentiation (CK14 and Involucrin expression) and their morphologic character of the clone.Results The percentages of CD+44 cells were about49.8% -53.5% and the median was 51.3%.After culturing CD+44 cells isolated from laryngeal carcinoma could proliferate and the percentage of CD+44 remained the same.CD+44 tumor cells contained much less RNA, more GO/GI cells, expressed more CK14 protein and less lnvolucrin protein (less differentiated state).CD+44 cells were muhangular in shape with protuberances; CD-44celis showed a sharp and spindle feature.In comparison with CD-44 cells, CD+44 cells could create heterogeneous offspring by single cell culture of limiting dilution.By observing clone forming rate after single cell planting, it was found that the CD+44 cells had stronger proliferation ability.Conclusions CD+44 cells possess some characteristics of stem cells, laryngeal carcinoma stem cells maybe exist in CD+44 cells.

Craniofacial Dysostosis ; Humans ; Receptors, Fibroblast Growth Factor

OBJECTIVETo study the effect of gene expression of mouse uroplakin II (UPII) promoter on human bladder cell cancer cell line.

METHODSThe mRNA expression of different cell lines was quantified by RT-PCR. Green fluorescent protein (GFP) and luciferase (Luc) were used as reporter genes. The plasmids carrying UPII or GFP were constructed and transfected into human cell lines of bladder transitional cell cancer (BIU-87), kindey cancer (GRC-1), vascular endothelium (EC), lung cancer cell line (A549) and skin fibroblast cell line (Hs27). GFP activity of cells was detected by confocual microscopy and flow cytometry (FCM). Luciferase value was measured by luminometer (microplate) and luciferase to beta-galactosidase ratios (L/G values) were used for evaluating transfection efficiency.

RESULTSRT-PCR showed high expression level of UPII mRNA in bladder cancer cell line BIU-87, whereas low level or no expression in nonbladder cancer cell lines. The activity of GFP in bladder cancer (BIU-87) cell was higher than that in the other cell lines (5 - 10/HP versus 0 - 2/HP), with 4.34% positive cells in BIU-87 detected by FCM, but no positive cell was found in the other cell lines. L/G values indicated that the luciferase expression in human bladder cancer cells transfected with mouse UPII promoter was 1.8 - 8.2-fold as high as that in the nonbladder cell lines.

CONCLUSIONMouse UPII promoter gene can be expressed in a tissue-specific fashion in human urinary bladder cancer. It is capable of initiating transcription of reporter genes in human bladder cancer cell line.

Animals ; Cell Line, Tumor ; Flow Cytometry ; Genetic Therapy ; Green Fluorescent Proteins ; Humans ; Luminescent Proteins ; genetics ; Membrane Proteins ; genetics ; Mice ; Organ Specificity ; Promoter Regions, Genetic ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; Urinary Bladder Neoplasms ; genetics ; therapy ; Uroplakin II

OBJECTIVETo separate the cell subpopulation with high tumorigenic ability and study the biological characteristics of this subpopulation in laryngeal carcinoma cells.

METHODSHuman laryngeal carcinoma cells were obtained by primary tissue culture technique. CD44 and CD133 molecules were used as markers to isolate the CD44(+), CD133(+), CD44(+)CD133(+) and CD44(+)CD133(-) cell subpopulations from the laryngeal carcinoma cells by flow cytometry. A nude mouse tumor xenograft model was developed for the study of the tumorigenic effects of the different cell populations. 1 x 10(6), 1 x 10(5), 1 x 10(4) and 1 x 10(3) cells were injected into the left flank of the mice, respectively. The mice were observed for palpable tumor formation and were sacrificed at 4 weeks later to assess the tumor formation rate, tumor volume and tumor weight. Boyden chamber migration assay was used to determine the migration ability and immunochemistry was used to detect the expression of stem cell antigen SCA-1 and beta1-integrin. Semi-quantities RT-PCR and Western blot analysis were performed to detect the expression level of Bmi-1 in the different cell subpopulations.

RESULTSThe growth of subcutaneous tumors in nude mice showed that a tumor can be generated with 1 x 10(3) CD44(+)CD133(+) cells. When the same dose of 1 x 10(6) CD44(+)CD133(+) cells was injected into the mice, both the average weight and volume of the tumors were significantly higher than those generated from other cell subpopulations. Boyden chamber migration assay showed that the invasion ability of CD44(+)CD133(+) cells was significantly higher than that of other cell subsets. The results of immunochemical analysis showed an abundant expression of stem cell antigen SCA-1 and beta1-integrin in the CD44(+)CD133(+) cells. Semi-quantitative RT-PCR and Western blot analysis provided strong evidence that the Bmi-1 expression in CD44(+)CD133(+) and CD133(+) cells was very significantly higher than that in CD44(+), CD44(+)CD133(-) and control cells (P < 0.01).

CONCLUSIONOur findings demonstrate that CD44(+)CD133(+) subset cells in laryngeal carcinoma posses some biological characteristics of tumor stem cells, which may be the original cells of laryngeal carcinoma and may become a new target of tumor therapy.

AC133 Antigen ; Animals ; Antigens, CD ; analysis ; Antigens, Ly ; metabolism ; Cell Adhesion ; Gene Expression Regulation, Neoplastic ; Glycoproteins ; analysis ; Humans ; Hyaluronan Receptors ; analysis ; Integrin beta1 ; metabolism ; Laryngeal Neoplasms ; immunology ; metabolism ; pathology ; Male ; Membrane Proteins ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Nuclear Proteins ; genetics ; metabolism ; Peptides ; analysis ; Polycomb Repressive Complex 1 ; Proto-Oncogene Proteins ; genetics ; metabolism ; RNA ; metabolism ; Repressor Proteins ; genetics ; metabolism ; Tumor Burden ; Tumor Cells, Cultured

BACKGROUNDMounting evidence suggests that tumors are histologically heterogeneous and are maintained by a small population of tumor cells termed cancer stem cells. CD133 has been identified as a candidate marker of cancer stem cells in laryngeal carcinoma. This study aimed to analyze the chemoresistance of CD133(+) cancer stem cells.

METHODSThe response of Hep-2 cells to different chemotherapeutic agents was investigated and the expression of CD133 was studied. Fluorescence-activated cell sorting analysis was used to identify CD133, and the CD133(+) subset of cells was separated and analyzed in colony formation assays, cell invasion assays, chemotherapy resistance studies, and analyzed for the expression of the drug resistance gene ABCG2.

RESULTSAbout 1% - 2% of Hep-2 cells were CD133(+) cells, and the CD133(+) proportion was enriched by chemotherapy. CD133(+) cancer stem cells exhibited higher potential for clonogenicity and invasion, and were more resistant to chemotherapy. This resistance was correlated with higher expression of ABCG2.

CONCLUSIONSThis study suggested that CD133(+) cancer stem cells are more resistant to chemotherapy. The expression of ABCG2 could be partially responsible for this. Targeting this small population of CD133(+) cancer stem cells could be a strategy to develop more effective treatments for laryngeal carcinoma.

AC133 Antigen ; ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; genetics ; metabolism ; Antigens, CD ; genetics ; metabolism ; Antineoplastic Agents ; pharmacology ; Blotting, Western ; Carcinoma ; genetics ; metabolism ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Flow Cytometry ; Fluorouracil ; pharmacology ; Glycoproteins ; genetics ; metabolism ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; Neoplasm Proteins ; genetics ; metabolism ; Neoplastic Stem Cells ; cytology ; drug effects ; metabolism ; Paclitaxel ; pharmacology ; Peptides ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVENucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes.

METHODSEukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results.

RESULTThe recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours.

CONCLUSIONThe nucleic vaccine constructed pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma.

Cancer Vaccines ; biosynthesis ; Gene Expression ; Genetic Vectors ; HN Protein ; genetics ; Hep G2 Cells ; Humans ; Interleukin-18 ; genetics ; Laryngeal Neoplasms ; immunology ; prevention & control ; Newcastle disease virus ; immunology ; Plasmids ; Transfection ; Vaccines, DNA ; biosynthesis

BACKGROUNDChronic active Epstein-Barr virus infection (CAEBV) has been previously reported to be sometimes associated with an aggressive clinical course. The characteristics of CAEBV in Mainland Chinese pediatric patients are largely unreported. The main aims of this survey were to recognize the clinical features of CAEBV in children and to explore its diagnostic criteria and risk factors.

METHODSA retrospective study was performed on 53 pediatric patients (36 boys and 17 girls) with CAEBV who were admitted to Beijing Children's Hospital between 2003 and 2007. All their medical records were reviewed and analyzed. For each patient, demographic, clinical, laboratory data and outcome were collected. Independent-samples t test was used for statistical analysis.

RESULTSThe age at onset of CAEBV was from 2 months to 14.6 years (mean (5.3+/-3.3) years). At the time of onset, 43.4% patients had an infectious mononucleosis-like symptom. Most patients exhibited intermittent fever (92.5%, 49/53), hepatomegaly (81.1%, 43/53) and splenomegaly (77.4%, 41/53). Life-threatening complications including hemophagocytic syndrome (24.5%, 13/53), interstitial pneumonia (24.5%, 13/53), hepatic failure (15.1%, 8/53) and malignant lymphoma (11.3%, 6/53) were also observed. The serum EBV DNA level in 23 patients with CAEBV was in the range of 5.05 x 10(2)-4.60 x 10(6) copies/ml with a mean value of 10(3.7) copies/ml. Many patients with CAEBV generally had continuous symptoms during the observational period. Eleven out of 42 patients (26.2%) died 7 months to 3 years after onset. Deceased patients were more likely to have had lower platelet counts and albumin levels than the living patients (P<0.05 for all comparisons).

CONCLUSIONSThe study reveals that CAEBV in Chinese pediatric patients has a severe clinical course and prognosis is poor. Thrombocytopenia and decreases in albumin might potentially be risk factors for a poor prognosis. EBV loads should be measured and tissue should be stained on hybridization probes for EBV-encoded small RNA (EBER) if a patient presents with the known symptoms of CAEBV.

Adolescent ; Age Distribution ; Child ; Child, Preschool ; China ; epidemiology ; Chronic Disease ; Epstein-Barr Virus Infections ; diagnosis ; epidemiology ; etiology ; pathology ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Risk Factors ; Serum Albumin ; analysis ; Thrombocytopenia ; complications

OBJECTIVETo establish decision tree and logistic regression classification models for diagnosing pancreatic adenocarcinoma (PaCa) and for screening serum biomarkers related to evaluation of different stages and curative effects.

METHODSSerum samples obtained from subjects with pancreatic adenocarcinoma (n = 58) and normal pancreas (n = 51) were applied to strong anion exchange chromatography (SAX2) chips for protein profiling by SELDI-TOF-MS to screen multiple serum biomarkers. Biomarker Wizard software and several statistical methods including algorithm of decision tree, logistic regression and ROC curves were used to construct the decision tree or logistic regression classification models.

RESULTSAverage of 61 mass peaks were detected at the molecular range of 2000-30,000, ten decision trees with the highest cross validation rate were chosen to construct the classification models, which can differentiate PaCa from normal pancreas with a sensitivity of 83.3% and a specificity of 100%. Logistic regression was used to achieve the AUC (0.976 +/- 0.011, P < 0.001) with a sensitivity of 77.6% - 91.4% and a specificity of 92.2% - 100%. Six mass peaks were combined by logistic regression to achieve the AUC 0.897 +/- 0.054, 0.978 +/- 0.021 and 0.792 +/- 0.107 (P < 0.05) in the three groups (patients at stage I and II, stage II and III, stage III and IV). One mass peak (M/Z 4,016) was screened (P < 0.05) significantly between the preoperative and postoperative PaCa samples and the intensity decreased weeks after operation.

CONCLUSIONDecision tree and logistic regression classification models of the mass peaks screened by SELDI-TOF-MS serum profiling can be used to differentiate pancreatic adenocarcinoma from normal pancreas, and is superior to CA 199. The detected mass peaks are helpful for the evaluation of curative effect and prognosis of pancreatic adenocarcinoma.

Adenocarcinoma ; blood ; diagnosis ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Area Under Curve ; Biomarkers, Tumor ; blood ; Blood Proteins ; analysis ; Chromatography, Ion Exchange ; methods ; Decision Trees ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; blood ; diagnosis ; pathology ; surgery ; Prognosis ; Protein Array Analysis ; Proteomics ; ROC Curve ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization