Administration, Oral ; Adolescent ; Female ; Histiocytosis, Langerhans-Cell ; complications ; drug therapy ; pathology ; Humans ; Lymph Nodes ; pathology ; Prednisone ; administration & dosage ; therapeutic use ; Retrospective Studies ; Skin Ulcer ; drug therapy ; etiology ; pathology ; Thalidomide ; administration & dosage ; therapeutic use ; Treatment Outcome

BACKGROUNDMitochondrial DNA 4977-bp deletion (DeltamtDNA(4977)) was reported in many human neoplasia. However, its biological significance remains to be evaluated and the molecular mechanism needs to be investigated. In this study, we analyzed the frequency of DeltamtDNA(4977) in gastric cancer (GC) cell lines and tissues, as well as reactive oxygen species (ROS) contents and manganese superoxide dismutase (MnSOD) expression levels in GC cell lines to explore its biological significance and molecular mechanism.

METHODSSemi-quantitative PCR and real-time PCR were used to detect the incidence of DeltamtDNA(4977) in 13 GC cell lines and 272 human gastric tissues (108 GC specimens and the respective adjacent normal tissues, and 56 normal gastric mucosa from non-cancer patients). We further identified intracellular ROS production by flow cytometry and MnSOD expression by semi-quantitative reverse transcription-PCR (RT-PCR) and Western blotting. Statistical analyses were carried out using the Logistic regression analysis and Kaplan-Meier method.

RESULTSBased on our earlier study, we optimized the PCR amplification condition by reducing the cycle number. In this study, we systematically documented the high incidence of DeltamtDNA(4977) in GC cell lines (10/13, 76.9%), GC tissues (86/108, 79.6%), matched normal tissues (73/108, 67.6%), and normal gastric mucosa of non-cancer patients (29/56, 51.8%). A significantly higher incidence of mutated DeltamtDNA(4977) was observed in GC tissues with respect to the adjacent normal tissues (79.6% vs 67.6%, P = 0.045), and they were both higher than that in normal controls (P < 0.05). Most importantly, we linked the DeltamtDNA(4977) mutations with the expression level of MnSOD and ROS contents. The cell lines containing lower expression level of MnSOD was found to have generally higher frequent DeltamtDNA(4977) and more ROS.

CONCLUSIONThe decreased anti-oxidative ability, which leads to increased ROS contents, is correlated with the mtDNA damage during gastric carcinogenesis.

Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Cell Line, Tumor ; DNA, Mitochondrial ; genetics ; Female ; Humans ; In Vitro Techniques ; Male ; Middle Aged ; Mutation ; Reactive Oxygen Species ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; genetics ; metabolism ; Superoxide Dismutase ; genetics ; metabolism

OBJECTIVETo evaluate the prognostic significance of various clinicopathologic parameters in gastrointestinal stromal tumor (GIST), and to study the frequency of c-kit exon 11 mutations in this tumor.

METHODSOne hundred and fifty-six cases of gastric or small intestinal GIST were retrieved from the archival files of the Department of Pathology, Chinese PLA General Hospital. The clinical features, site of occurrence, tumor diameter, mitotic index, coagulative tumor necrosis, and risk grade were studied and analyzed statistically. Tumor DNA was extracted and c-kit exon 11 was amplified. Upon detection by denaturing high-performance liquid chromatography, the amplified exon 11 was sequenced.

RESULTSFor the 83 cases of gastric GIST studied, the mean age of patients was 55.4 years. Follow-up information was available in 62 cases, with 17 cases having local recurrence or distant metastasis. The 5-year survival rate was 66.5% +/- 17.1%. For the 73 cases of small intestinal GIST studied, the mean age of patients was 50.6 years. Follow-up information was available in 43 cases, with 22 cases having local recurrence or distant metastasis. The 5-year survival rate was 61.8% +/- 18.3%. In general, for gastric GIST, age younger than 50 years (P = 0.046), advanced clinical stage (P = 0.0001), large tumor size (P = 0.0001), high mitotic index (P = 0.0001), presence of coagulative tumor necrosis (P = 0.0001), and high risk grade (P = 0.004) were associated with lower survival rate. COX hazard proportional model revealed that advanced clinical stage (P = 0.001), large tumor size (P = 0.001), high mitotic index (P = 0.002) and high risk grade (P = 0.018) indicated worse prognosi. For small intestinal GIST, advanced clinical stage (P = 0.010) and presence of coagulative tumor necrosis (P = 0.036) were associated with lower survival rate. Advanced clinical stage was an independent prognostic factor. A total of 25 cases harbored c-kit mutations. The frequency of c-kit mutations was 32% and 22.5% for gastric and small intestinal GIST respectively. For gastric GIST, c-kit mutations occurred mainly in patients older than 50 years. In contrast, c-kit mutations in small intestinal GIST occurred in the age group of 40 to 49 years.

CONCLUSIONSFor gastric GIST, advanced clinical stage, tumor diameter, mitotic index and risk grade are the main prognostic indicators. For small intestinal GIST, advanced clinical stage and presence of coagulative tumor necrosis indicate poor prognosis. In general, small intestinal GIST is more frequently associated with metastasis and tumor relapse than gastric GIST. The occurrence of c-kit mutations also correlates with age of patients.

Adolescent ; Adult ; Age Factors ; Aged ; Aged, 80 and over ; Bone Neoplasms ; secondary ; DNA, Neoplasm ; genetics ; Disease-Free Survival ; Exons ; Female ; Follow-Up Studies ; Gastrointestinal Stromal Tumors ; genetics ; pathology ; Humans ; Liver Neoplasms ; secondary ; Male ; Middle Aged ; Mutation ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Proportional Hazards Models ; Proto-Oncogene Proteins c-kit ; genetics ; Survival Rate ; Tumor Burden ; Young Adult

Apoptosis ; drug effects ; Arachidonate 12-Lipoxygenase ; analysis ; physiology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Enzyme Inhibitors ; pharmacology ; Flavanones ; pharmacology ; Humans ; Lipoxygenase Inhibitors ; Multiple Myeloma ; drug therapy ; pathology

OBJECTIVETo apply the functionally generated path (FGP) technique for modeling the occlusal surface of a computer-aided-design (CAD) posterior full crown to obtain anatomic morphology.

METHODSA patient with defected left mandibular first molar was employed. After tooth preparation and impression making, the gypsum working cast and die were scanned with a digitized mechanical scanner and the surface data was acquired. The interocclusal records at intercuspal position (ICP) and FGP were made in the patient's mouth. These records were placed on the working cast and their surfaces were scanned. In the process of the computer-aided designing full crown, the cusps and fossae of the occlusal surfaces were accurately modified according to the digitized information of ICP and FGP interocclusal records.

RESULTSA full crown was designed and the occlusal morphology of the restoration was adapted to dynamic occlusion as well as static occlusion.

CONCLUSIONSThe FGP technique was practical for the CAD of full crown and could avoid potential occlusal interferences with opposing teeth during function.

Adult ; Bite Force ; Computer-Aided Design ; Crowns ; Dental Occlusion ; Dental Prosthesis Design ; methods ; Humans ; Male

OBJECTIVETo study gene expression of collagen types IX and X in human lumbar intervertebral discs during aging and degeneration and to explore the role of collagen types IX and X in disc degeneration.

METHODSFetal, adult and pathologic specimens were subjected to in situ hybridization with cDNA probes to investigate mRNA-expressions of types IX and X collagen gene.

RESULTSIn fetal intervertebral discs, positive mRNA hybridization signals of type IX collagen were concentrated in the nucleus pulposus and the inner layer of anulus fibrosus. Interstitial matrix of the nucleus pulposus also showed positive type X collagen staining. Positive mRNA hybridization signals of types IX and X were not detected in the middle and outer layers of anulus fibrosus. In adult specimens, expression of type IX collagen mRNA was markedly decreased. No hybridization signals of type X collagen was observed. As for pathological specimens, there was no gene expression of type IX collagen. In severe degenerated discs from adults, there were focal positive expressions of type X collagen.

CONCLUSIONSObvious changes of collagen gene expression occur with aging. Expression of type IX collagen decreases in adult and pathological discs. Results of type X collagen expression suggest that type X collagen is expressed only in older adult and senile discs (i.e., when disc degeneration has already reached a terminal stage), indicating the terminal stage of degeneration.

Adolescent ; Adult ; Collagen Type IX ; metabolism ; Collagen Type X ; metabolism ; Female ; Gene Expression ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Intervertebral Disc ; embryology ; metabolism ; Lumbar Vertebrae ; Male

This study was aimed to observe the effects of siRNA on Livin expression and function in K562 cells. Livin siRNA were designed and synthesized, then were transfected into K562 cells by using AMAXA nucle transfactor. Expressions of Livin mRNA and protein in transfected K562 cells was detected by RT-PCR and Western blot respectively. Non-transfected cells were used as control. The enhanced green fluorescent protein plasmid was used as positive control and the transfection efficiency was detected by flow cytometry. Cell apoptosis was measured by flow cytometry with Annexin V-FITC/PI double staining. The results showed that the transfection efficiency of electroporation method was about 50. The synthesized siRNA inhibited livin expression at both mRNA and protein levels. The rate of K562 cell apoptosis increased from (9.63 ± 0.89) in control group to (12.07 ± 1.39) and (27.41 ± 2.30) at 24 h and 48 h after transfection, respectively (P < 0.05). It is concluded that the siRNA can inhibit anti-apoptosis of livin gene via down-regulating livin gene expression, which may provide the new method for anti-leukemia study.
Adaptor Proteins, Signal Transducing ; genetics ; Apoptosis ; genetics ; Gene Expression Regulation, Leukemic ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; K562 Cells ; Neoplasm Proteins ; genetics ; RNA, Small Interfering ; genetics

BACKGROUNDDysregulated metallothionein 2A (MT2A) has been implicated in carcinogenesis. The purpose of this study was to investigate the expression of MT2A in gastric cancer (GC) and its correlation with prognosis.

METHODSReverse transcription-polymerase chain reaction and real-time polymerase chain reaction were used to detect the mRNA expression of MT2A in 12 GC cell lines, normal gastric epithelial GES-1 cells, and 36 GC and adjacent normal tissues. MT2A protein expression was determined in 258 GC tissues and 171 adjacent normal tissues by immunohistochemistry.

RESULTSMT2A mRNA expression was lower in GC cells and primary tumors than in GES-1 cells and adjacent normal tissues, respectively. High protein expression of MT2A was present in 130 of 171 normal tissues (76.0%) and in 56 of 258 GC tissues (21.7%; P < 0.001). MT2A protein expression was higher in well/moderately differentiated GC (22/54; 40.7%) than in poorly differentiated GC (34/204; 16.7%; P < 0.001). Moreover, the protein expression of MT2A was lower in diffuse-type GC (6/82; 7.3%) than in intestinal-type GC (50/176; 28.4%; P = 0.0001). Importantly, MT2A expression was an independent prognostic factor for GC, and decreased MT2A expression was associated with poor clinical outcome (P < 0.001). The expression status of MT2A could predict prognosis in intestinal and diffuse-type GCs.

CONCLUSIONExpression status of MT2A might be a useful prognostic biomarker for GC, especially when used in combination with Lauren's classification.

Adult ; Aged ; Cell Line, Tumor ; Female ; Humans ; Logistic Models ; Male ; Metallothionein ; analysis ; genetics ; MicroRNAs ; analysis ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Stomach Neoplasms ; chemistry ; classification ; pathology

OBJECTIVETo study the possible clonal origin of neuroendocrine cells in colorectal adenocarcinoma.

METHODSTwenty-six microsatellite loci were screened using laser capture microdissection, DNA extraction and whole genome amplification. Microsatellite instability (MSI) and loss of heterozygosity (LOH) in adenocarcinoma cells and neuroendocrine cells amongst 30 cases of colorectal carcinoma with neuroendocrine differentiation were detected using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP)-silver staining. The mutation status of p53 was evaluated by PCR-sequencing. The clonal origin of neuroendocrine cells in colorectal adenocarcinoma was determined.

RESULTSAmongst the 30 cases studied, the prevalence of MSI was 16.9% while that of LOH was 8.5%. The rate showed no statistically significant difference between adenocarcinoma cells and neuroendocrine cells. In 6 cases, the microsatellite alteration was entirely consistent. In 23 cases, the rate of microsatellite alteration consistency was greater than that of inconsistency. In 1 case, the consistency and inconsistency rates were identical. There was statistically significant difference between consistency and inconsistency of microsatellite alteration. The prevalence of p53 mutation was 16.7% which was the same for both adenocarcinoma cells and neuroendocrine cells.

CONCLUSIONSAdenocarcinoma cells and neuroendocrine cells in colorectal adenocarcinoma with neuroendocrine differentiation have similar biologic changes. It is likely that they are of identical origin.

Adenocarcinoma ; genetics ; pathology ; Colorectal Neoplasms ; genetics ; pathology ; DNA Mutational Analysis ; Humans ; Laser Capture Microdissection ; Loss of Heterozygosity ; Microsatellite Instability ; Neuroendocrine Cells ; pathology ; Tumor Suppressor Protein p53 ; genetics

OBJECTIVETo evaluate the efficacy of hBMP-4 gene modified tissue engineered bone graft in the enhancement of rabbit spinal fusion and find an ideal kind of substitute for the autograft bone.

METHODSRabbit BMSCs were cultured and transfected with AAV-hBMP-4 using different MOI value. The optimal MOI value were determined by observing cell's morphology change. BMSCs were then transfected with AAV-hBMP4 and AAV-EGFP respectively, following which the transfected cells were evenly suspended in a collagen sponge I, and implanted to either side of the L5,6 intertransverse spaces posterolateral in the New Zealand rabbits to induce spinal fusion. Fourteen rabbits were randomly divided into 2 groups. Group 1: AAV-hBMP-4 transfected BMSCs in the right side (hBMP-4 side) and autograft bone in the left side. Group 2: AAV-hBMP-4 transfected BMSCs in the right side (hBMP-4 side) and AAV-EGFP transfected BMSCs in the left side (EGFP side). Radiographs and three-dimensional CT of the spine, manual palpation, gross and histological examination of the fusion masses for all the animals were performed subsequent to animals having been sacrificed at 12 weeks after surgery.

RESULTSEvaluation has been taken in 12 New Zealand rabbits delivered into 2 groups which meet the criterion after operation. Eleven in 12 implemented sides involved hBMP-4 achieved bony fusion, to which 5 in 6 autografted sides was similar. But only 2 in 6 sides in EGFP-group achieved bony fusion meanwhile. Three-dimensional CT scan and palpation also evidenced the results. Bone formation was observed obviously on specimen both in hBMP4 sides and autografted ones. EGFP-group also got bony integration, but the quantity was small.

CONCLUSIONTissue-engineered bone graft constructed from application of hBMP4 is a fine substitute for autograft. Effective enhancement of bony integration in spinal fusion surgery has been evidenced in vivo.

Animals ; Bone Morphogenetic Protein 4 ; genetics ; Bone Regeneration ; Bone Substitutes ; Bone Transplantation ; methods ; Genetic Vectors ; Lentivirus ; genetics ; Male ; Myeloid Progenitor Cells ; Rabbits ; Random Allocation ; Spinal Fusion ; methods ; Stromal Cells ; Tissue Engineering ; Transfection