Cannula ; Catheters ; Drainage ; Duodenal Diseases ; Humans ; Intestinal Fistula/surgery*

OBJECTIVETo use a glycemic method to screen hepatocellular carcinoma (HCC) metastasis related aberrant 1-6 fucosylated glycoproteins, and to analyze the metastasis-related alterations of annexin II.

METHODS2-DE coupled with lectin affinity blot, lectin affinity precipitation followed by MALDI-TOF-MS/MS was established to screen glycoproteins related to HCC metastasis. Immunofluorescence analysis, Western blot and real-time PCR were performed on higher and lower metastatic HCC cell lines to detect the protein expression levels and mRNA levels of annexin II.

RESULTSThe Lens culinaris agglutinin (LCA) affinity glycoprotein profiles from MHCC97-L and MHCC97-H cells were differentially displayed when compared with Hep3B. Annexin II was identified by MALDI-TOF-MS/MS and its increased core-fucosylation was associated with HCC metastasis and it was confirmed. In addition, we found that annexin II was distributed in the cytoplasm and it had higher protein and gene expressions in MHCC97-L and MHCC97-H cells than in Hep3B cells.

CONCLUSIONOur results suggest that the increase of annexin II and its expression levels, and the increase of core-fucosylation might all be related to HCC metastatic ability.

Annexin A2 ; analysis ; genetics ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Electrophoresis, Gel, Two-Dimensional ; Gene Expression ; Humans ; Neoplasm Metastasis ; Neoplasm Proteins ; analysis ; genetics

OBJECTIVETo explore the expression of X-linked inhibitor of apoptosis protein (XIAP) in Tca8113 cell, and to investigate its relationship to the chemoresistance. METHODS; The Tca8113 cell line was cultured by IMDM and the concentration of Pingyangmycin (PYM) added to Tca8113 cell line was increased gradually and continually, which was to induce the PYM-resistance in Tca8113 cell line. The sensitivity of Tca8113 cell to PYM and expression of XIAP were measured with methyl thiazolyl tetrazolium (MTT) chromatometry and reverse transcription-polymerase chain raction (RT-PCR). The XIAP level in the cells and its chemoresistance to PYM were analyzed by linear regression.

RESULTSThe IC50 of Tca8113-1-10 group and Tca8113-10-10 group were(12.758 +/- 0.030), (18.986 +/- 0.150) microg x mL(-1) respectively. The IC50 of Tca8113-1-20 group and Tca8113-10-20 group increased to (26.302 +/- 0.072), (35.294 +/- 0.115) microg x mL(-1) respectively. There was a relation between XIAP and the drug-resistance in Tca8113 cell.

CONCLUSIONXIAP may play an important role in the chemoresistance which might serve as a new therapeutic target for oral squamous cell carcinoma.

Apoptosis ; Bleomycin ; analogs & derivatives ; Carcinoma, Squamous Cell ; Cell Line, Tumor ; Drug Resistance, Neoplasm ; Humans ; X-Linked Inhibitor of Apoptosis Protein

Objective: To establish a method for isolation and purification of fetal membrane derived adherent cells (FMDACs) , and investigate their biological characteristics. Method: FMDACs were isolated with trypsin inducing and cultured in vitro. FMDACs were induced to differentiate into osteoblasts and adipocytes. FACS and immunocytochemistry technique were used to examine the cell surface antigen. The genetic stability was verified by karyotype analysis. Results: FMDACs were successfully isolated and expanded in vitro. They had strong proliferative ability. FMDACs were positive for CD44 and CD29, but negative for CD34, CD14 and CD45. FMDACs were differentiated into osteoblasts and adipocytes after inducement. The karyotype was stable in the sixth-passaged FMDACs and the tumorigenicity was not found. Conclusion; FMDACs have the possibility of multipotent stem cells, which have strong capacities of self-renewal and multidirectional differentiation. The genetic background of FMDACs is stable. FMDACs may be used as a kind of novel seed cells for tissue engineering.

OBJECTIVETo screen low molecular weight protein biomarkers relevant to portal vein tumor thrombi (PVTT) in serum of hepatocellular carcinoma (HCC) patients.

METHODSSerum samples were obtained from 12 healthy volunteers, 12 HCC patients without PVTT and 12 HCC patients with PVTT. Using two-dimensional gel electrophoresis (2-DE) in which the second dimension was 16% SDS-PAGE, serum protein images of the 3 groups were analyzed by ImageMaster software. The differential protein spots were further identified by MALDI-TOF MS/MS.

RESULTSComparing the results using 12.5% SDS-PAGE gel, there were more protein bands (between 3 x 10(3) and 20 x 10(3)) and low molecular weight (MW) protein spots (less than 20 x 10(3)) were clearly shown in the 16% SDS-PAGE gel. Fifteen differential protein spots representing 5 proteins were found in the 3 groups by inter-class comparison and they were then identified. Compared with those in the healthy group, apolipoprotein A-I, lipoprotein CIII, transthyretin and DNA topoisomerase II were all down regulated in HCC groups and haptoglobin-2 was over expressed. All 5 proteins decreased more in the PVTT group than in the non-PVTT group.

CONCLUSIONThe expression of low MW serum protein obviously changes in the beginning and in the progressive stage of HCC, and differentially expressed low MW proteins might be potential biomarkers in an early prognostic prediction and surveillance in the treatment for HCC and PVTT.

Adult ; Blood Proteins ; analysis ; Carcinoma, Hepatocellular ; blood ; pathology ; Electrophoresis, Gel, Two-Dimensional ; methods ; Female ; Humans ; Liver Neoplasms ; pathology ; Male ; Middle Aged ; Neoplastic Cells, Circulating ; pathology ; Portal Vein ; pathology ; Proteome ; analysis

OBJECTIVETo evaluate the mRNA and protein expressions of peroxiredoxin II (PrxII) in hepatocellular carcinoma (HCC) and their significance.

METHODSHCC was induced by aflatoxin B1 (AFB1) in 6 tree shrews (Tupaia belangeri chinensis). The expression levels of PrxII mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot on HCC tissues and on their surrounding liver tissues (para-HCC). Biopsied liver tissues were taken before the HCC induction (pre-HCC) from the same animals and from a group of blank controlled animals that served as controls. Liver biopsy specimens from 18 cases of human HCC and from 17 healthy human volunteers were studied using the same methods.

RESULTSThe mRNA and protein expressions of PrxII in tree shrew HCC tissues were significantly higher than those in para-HCC and pre-HCC tissues, and also higher than those in the liver tissues from the control animals (all P < 0.05). The expression levels of PrxII mRNA and protein in human HCC tissues were also significantly higher than those in their para-HCC tissues and in the human normal liver tissues (P < 0.05).

CONCLUSIONPrxII might play an important role in hepatocarcinogenesis and might be used as a molecular target for HCC prevention and treatment.

Adult ; Aged ; Animals ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Female ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Liver Neoplasms, Experimental ; metabolism ; pathology ; Male ; Middle Aged ; Peroxiredoxins ; genetics ; Tupaiidae

OBJECTIVE: To investigate the source of the extra small chromosome in a patient with karyotype 45,X[115]/46,X + mar[45]/46,XY[29]. METHODS: The SRYgene was detected by PCR, and the chromosome Y probe that labeled with biotin was detected by fluorescence in situ hybridization. RESULTS: SRY gene is detected positive and the mar chromosome showed positive signal with FISH in human chromosome Y probe pool. CONCLUSION The extra small chromosome is part of the chromosome Y.
Adolescent ; Chromosome Aberrations ; Female ; Genes, sry ; genetics ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping ; Polymerase Chain Reaction ; Sex Chromosomes ; Sex Differentiation ; genetics ; Turner Syndrome ; genetics

OBJECTIVE: To identify the gene causing diffuse palmoplantar keratoderma in a Chinese pedigree. METHODS: Four normal individuals and 3 patients in a diffuse palmoplantar keratoderma family and 10 unrelated control samples were recruited. The hotspot of the mutations of keratin 9 gene was analyzed by polymerase chain reaction and direct sequencing. RESULTS: We found a G485A transition in ke ratin 9 gene, resulting in the substitution of glutamine for arginine at codon 162 in this diffuse palmoplantar keratoderma family. The mutation was not found in the 10 unrelated control samples and 4 normal individuals. CONCLUSION The mutation G485A found in keratin 9 gene is the disease-causing mutation in the diffuse palmoplantar keratoderma family.
Base Sequence ; DNA Mutational Analysis ; Female ; Heterozygote ; Humans ; Keratins ; genetics ; Keratoderma, Palmoplantar, Diffuse ; genetics ; Male ; Molecular Sequence Data ; Mutation ; Pedigree

OBJECTIVE: To investigate the biological characteristics of endothelial progenitor cells (EPCs) from the umbilical cord blood (UCB), and to evaluate their oncogenicity after long-term culture in vitro. METHODS: The mononuclear cells (MNCs) were isolated from the UCB and cultured in MCDB131 medium supplemented with 20% FBS, VEGF and other growth factors. Morphology of the EPCs was observed, and the growth curve of the EPCs was investigated. Surface antigens of the EPCs were analyzed by the flow-cytometer. The capability of intaking the acetylated low-density lipoprotein (acLDL) of the EPCs was detected using fluoresencent chemical method. The vasoformative capability and genetic stability of EPCs were cultured in matrigel, and examined by karyotype analysis. The oncogenicity of EPCs was verified by the tumorigenesis test in athymic mouse and soft agar. RESULTS: EPCs were successfully derived from the UCB, and could be passaged to at least 42(nd) generation and had strong abilities of proliferation, acLDL intake and vasoformation, but there was not oncogenicity. They expressed endothelial cell-surface antigens and maintained normal karyotype. CONCLUSION The EPCs with proliferative potential can be isolated from the UCB. They can be passaged in long-term cultures without oncogenicity, and can maintain normal karyotype. The EPCs can be served as a new type of cells in cell and gene therapy.
Animals ; Antigens, Surface ; analysis ; Cell Line ; Cell Proliferation ; drug effects ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Fetal Blood ; cytology ; Flow Cytometry ; HeLa Cells ; Humans ; Infant, Newborn ; Intercellular Signaling Peptides and Proteins ; pharmacology ; Karyotyping ; Mice ; Mice, Nude ; Neoplasms, Experimental ; pathology ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVE: To identify the origin of the marker chromosome in a patient with chromosome aberration, and to provide the precise genetic diagnosis. METHODS: Comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH) were performed to detect the known small marker chromosome in this patient. RESULTS: The small marker chromosome originated from chromosome 13 pter->q12. CONCLUSION CGH and FISH can be used to detect the small marker chromosome, which is convenient and quick in detecting the origin of small marker chromosome.
Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 13 ; genetics ; Female ; Genome, Human ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Karyotyping ; Nucleic Acid Hybridization ; methods