Actins ; metabolism ; Bone Neoplasms ; blood ; complications ; diagnostic imaging ; pathology ; surgery ; Diagnosis, Differential ; Female ; Humans ; Hypophosphatemia ; blood ; etiology ; Ilium ; Mesenchymoma ; blood ; complications ; diagnostic imaging ; pathology ; surgery ; Middle Aged ; Osteomalacia ; blood ; etiology ; Phosphates ; blood ; Platelet Endothelial Cell Adhesion Molecule-1 ; metabolism ; Tomography, X-Ray Computed

ObjectiveTo investigate the diagnostic value of EUS-FNA for pancreatic masses and correlated influential factors. MethodsWe retrospectively analyzed the clinical data of 101 patients with pancreatic lesions who underwent EUS-FNA from January 2008 to January 2010. The clinical data enrolled 10 factors including patient gender, patient age, lesion location, lesion size, lesion characteristics, negative suction pressure, times of access, real-time cytological diagnosis, type of EUS and operators' experiences.ResultsThe overall diagnostic accuracy, sensitivity, specificity, positive predictive value and negative predictive value of EUS-FNA were 85. 1％, 81.1％, 96. 3％, 98. 4％ and 65.0％, respectively. Univariable logistic regression analysis indicated that lesion size, lesion characteristics, negative suction pressure, operators' experience were correlated factors of EUS-FNA positive rate, while lesion size was the only correlated factor of EUS-FNA diagnostic accuracy ( OR =1. 984,95 ％ CI: 1. 141 ～ 3. 451, P =0. 015 ). Every 1 cm the lesion increased, by 1.67 times of opportunity the positive rate became, by 1.83 times of opportunity the accuracy was. The lesion size and lesion characteristics were independent correlated factors of EUS-FNA positive rate (OR=2.012, P=0.000; OR =10.218, P=0. 002). The positive rate of EUS-FNA in solid lesions was 10. 2 times of that in cystic lesions. Lesion size was the independent correlated factors of EUS-FNA diagnostic accuracy (OR =1. 984, P =0. 015 ). ConclusionEUS-FNA can effectively make a pathological diagnosis of pancreatic masses with high diagnostic accuracy and specificity. EUS-FNA diagnostic positive rate and accuracy were both positively correlated with pancreatic lesion size. EUS-FNA positive rate of solid pancreatic lesions is significantly higher than that of cystic lesions.

OBJECTIVETo study the histochemical staining in the diagnosis of osteosarcoma.

METHODSTo compare the effectiveness of picrosirius red, improved Ponceau trichrome and Masson trichrome staining methods on bone formation tissues in conventional osteosarcoma, paraosteal osteosarcoma, periosteal osteosarcoma, extraskeletal osteosarcoma, inflammatory myofibroblastic tumour, malignant fibrohistiocytoma, chondrosarcoma, fibrosis with ossification and calcification.

RESULTSWith modified Ponceau trichrome staining, bone formation tissues showed a homogenous, orange-red interblended with blue in color. From osteoid to mature bone the color changed from orange-red, light blue to dark blue. Fibrotic tissue was stained blue in color with striated appearance. Cartilage was not stained. Picrosirius red method gave bone formation tissues homogenous staining. Along with bone maturation, from osteoid tissue to mineralized bones, the color showed changes from light red, yellow, orange-red, red to dark purple. The cartilage demonstrated homogenous light red in color. Fibrous tissue stained red interblended with yellow in color, striated in shape. With Masson trichrome staining osteoid displayed pale blue and mineralized bone showed dark blue in color. Fibrotic tissue showed a striated blue staining.

CONCLUSIONThe modified Ponceau trichrome and Picrosirius red staining methods are better than Masson trichrome to demonstrate bone formation tissue in osteosarcoma. The former two methods could be also used in study on bone formation.

Bone Neoplasms ; pathology ; Histocytochemistry ; Humans ; Osteosarcoma ; pathology ; Staining and Labeling ; methods

OBJECTIVETo study the clinicopathologic features, diagnosis and differential diagnosis of myositis ossificans (MO).

METHODSThe clinical features, radiologic results and pathologic findings of 15 cases of MO (including biopsy and surgical specimens) were analyzed. The hematoxylin and eosin sections were reviewed under light microscope. Immunohistochemical staining for S-100 protein, vimentin, desmin, actin and osteonectin was performed.

RESULTSThe age of the patients ranged from 12 to 46 years. The male-to-female ratio was 11:4. Thirteen cases were located in the parosteum of long bone or subperiosteal soft tissue. The remaining two cases occurred in iliac region and palm, respectively. Five patients had history of injury, while 2 patients had operation before. Four patients had no history of trauma and the remaining one had unknown clinical history. Histologically, zonation pattern was not conspicuous in 10 biopsy cases and 8 corresponding surgical specimens. On the other hand, zonation pattern was observed in 5 biopsy cases and 7 corresponding surgical specimens. Follow up revealed relapses in two patients. Immunohistochemical study showed various degree of positivity for vimentin, desmin, actin and osteonectin. S-100 protein was focally positive in 2 of the cases. The Ki-67 index varied from 1% to 10%.

CONCLUSIONCorrect diagnosis of MO relies on correlation of clinical features, radiologic examination and pathologic findings.

Adolescent ; Adult ; Biopsy ; Child ; Female ; Humans ; Male ; Middle Aged ; Myositis Ossificans ; diagnosis ; genetics ; pathology ; S100 Proteins ; genetics ; Vimentin ; X-Rays ; Young Adult

OBJECTIVETo investigate the biological role of auto-induced expression of hepatitis C virus (HCV) core protein (protein C) using a recombinant protein in an in vitro cell-based system.

METHODSThe PCR-amplified full-length HCV protein C gene (573 bp) was inserted into the pET28a prokaryotic expression vector. The recombinant plasmid was transformed into BL21(DE3)pLysS E. coli to achieve high-concentration expression of the recombinant C protein by auto-induction. The recombinant protein C was purified by Ni-NTA affinity chromatography, and tested in a protein binding assay for its ability to bind the HCV NS3 protein.

RESULTSThe transformed E. coli produced a large amount of recombinant protein C, as detected in the sonicated supernatant of the bacteria culture. The antigenic reactivity of the recombinant protein C was confirmed by western blotting. However, the recombinant protein C could not be purified by Ni-NTA affinity chromatography, but co-precipitated with the HCV NS3 protein.

CONCLUSIONSoluble recombinant protein C was successfully expressed by auto-induction, and shown to interact with the HCV NS3 protein, which provides a novel insight into the putative biological activity of this factor in HCV-related molecular processes. Future studies of this recombinant HCV protein C's crystal structure and antigenicity may provide further clues to its biological function(s) and potential for clinical applications.

Escherichia coli ; metabolism ; Genetic Vectors ; Hepacivirus ; Recombinant Proteins ; genetics ; metabolism ; Viral Core Proteins ; biosynthesis ; genetics ; metabolism ; Viral Nonstructural Proteins ; metabolism

OBJECTIVESmith-Fineman-Myers syndrome (SFMS) is an X-linked mental retardation syndrome. The authors had ascertained a large Chinese family with SFMS from Shandong and had mapped the disease locus to an interval of 19.8 Mb on Xq25 flanked by markers DXS8064 and DXS8050. Further investigation suggested that SFMS exhibited locus heterogeneity. In this study for facilitating the identification of the gene responsible for SFMS, the additional markers were analyzed to narrow down the candidate region, and four candidate genes (GPC3, MST4,GPCR2 and GLUD2) were chosen and screened for disease-causing mutation.

METHODSPCR and denaturing polyacrylamide gel electrophoresis were used to genotype 13 new polymorphic markers distributed within the candidate region. Mutation detection was accomplished by sequencing the exons and intron-exon junctions of the candidate genes.

RESULTSBy analyzing 13 additional polymorphic markers, SFMS candidate region can be reduced to an interval of 10.18 Mb bounded by XSTR3 and XSTR4, and no disease-causing mutation was identified in the coding regions of four candidate genes.

CONCLUSIONGPCR2 GPC3, MST4 and GLUD2 were excluded as pathogenic genes for SFMS. The refined SFMS locus will assist in the identification and characterization of other candidate genes for SFMS.

Abnormalities, Multiple ; genetics ; Chromosome Mapping ; Chromosomes, Human, X ; Genetic Linkage ; Glutamate Dehydrogenase ; genetics ; Glypicans ; Humans ; Intellectual Disability ; genetics ; Male ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; Protein-Serine-Threonine Kinases ; genetics ; Receptors, G-Protein-Coupled ; genetics ; Syndrome

BACKGROUNDAlthough antiangiogenesis therapy plays an important role in anti-neoplastic treatment with its recognized efficacy and slight adverse effect, there is no prospective clinical trial to define ideal markers for predicting efficacy of antiangiogenic therapy. This study was undertaken to investigate the changes of activated circulating endothelial cells (aCECs) and survivin after anti-angiogenesis therapy and their significance in predicting the efficacy of the therapy.

METHODSPatients of non-small cell lung cancer (NSCLC) treated with chemotherapy with or without Endostar were observed. The amount of activated CECs was detected by flow cytometry, and the expression of survivin mRNA was determined by real-time polymerase chain reaction (PCR).

RESULTSAfter treatment, the amount of activated CECs decreased significantly in clinical benefit cases (P = 0.021 in chemotherapy alone, P = 0.001 in chemotherapy plus Endostar), increased in disease progressive cases (P = 0.015 in chemotherapy alone, but P = 0.293 in chemotherapy with Endotatar). After therapy, the expression of survivin mRNA decreased in clinical benefit cases (P = 0.001) and increased in disease progressive cases (P = 0.018). A positive correlation was found between activated CECs and survivin in the chemotherapy group pre- and post-therapy (P = 0.001 and 0.021, respectively), but only in the chemotherapy with Endostar group pre-therapy (P = 0.030) rather than post-therapy. A positive correlation was found between the decreased activated CECs after therapy and time to progression (TTP) (r = 0.322, P = 0.012); a negative correlation was found between the amount of survivin mRNA in serum post-therapy and TTP (r = -0.291, P = 0.048).

CONCLUSIONSActivated CECs and survivin may be ideal markers forecasting efficacy and prognosis of NSCLC. The former can reflect more sensitively antiangiogenic efficacy and the latter is more sensitive to shrinkage or swelling of tumors. Their combination can evaluate more accurately the efficacy of antiangiogenic therapy of NSCLC.

Angiogenesis Inhibitors ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; metabolism ; Endostatins ; therapeutic use ; Endothelial Cells ; drug effects ; metabolism ; Female ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Immunohistochemistry ; Inhibitor of Apoptosis Proteins ; Lung Neoplasms ; drug therapy ; genetics ; metabolism ; Male ; Microtubule-Associated Proteins ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo select short tandem repeats(STR) from X chromosome.

METHODSSTR is a universal genetic marker that has changeable polymorphism and stable heredity in human genome. It is a specific DNA segment composed of 2-6 base pairs as its core sequence. It is an ideal DNA marker used in linkage analysis and gene mapping. In this study, 8 short tandem repeats were selected from two genomic clones on X chromosome by using BCM Search Launcher. Primers amplifying the STR loci were designed by using Primer 3.0 according to the unique sequence flanking the STRs. Polymorphisms of the short tandem repeats in Chinese population were evaluated by PCR amplification and PAGE.

RESULTSFive of these STRs were polymorphic. Chi-square test indicated that the distribution of genotypes agreed with Hardy-Weinberg equilibrium (P>0.05).

CONCLUSIONFive polymorphic short tandem repeats have been identified on chromosome X and will be useful for linkage analysis and gene mapping.

Chromosomes, Human, X ; genetics ; Female ; Genotype ; Humans ; Microsatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics

OBJECTIVETo evaluate the feasibility of identifying oral pathogenic bacteria by comparing the metabolic profiling of putative periodontal pathogens and try to find a convenient and rapid way to discriminate oral microorganisms.

METHODSSuspensions of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum with same density were prepared and cultured respectively at liquid BHI medium. Then the growth quantity was measured periodically through turbidimetry and the growth curves of the inoculated bacteria were completed. The culture solutions of stable growth phase were sampled and characterized by 1H-nuclear magnetic resonance 1H-NMR). The data of 1H-NMR spectroscope results were analyzed by principal components analysis (PCA).

RESULTSThe PCA showed the obvious clustering phenomena and the points of three groups differentially centralized to three clusters. Therefore, the NMR-based metabonomics profiles could discriminate the three different kinds of bacteria.

CONCLUSIONThe metabonomics is a potential classable method to identify the oral pathogenic bacteria.

Aggregatibacter actinomycetemcomitans ; Bacteria ; Fusobacterium nucleatum ; Metabolomics ; Mouth ; microbiology ; Porphyromonas gingivalis ; Prevotella intermedia