Objective To investigate the effect of recombinant adenovirus mediatied human endostatin (rAD-GFP-ES)on rats with collagen typeⅡinduced arthritis(CIA),and explore the mechanism of inflamma- tion and cytokines inhibition on rats CIA.Methods The rAD-GFP-ES was amplified and purified.The model of rat CIA was induced by intradermal injection of typeⅡcollagen combined with complete Freund's adjuvant(CFA). On the second day after the injection,the therapeutic administration of rAD-GFP-ES(1?10~(11)pfu?kg~(-1)?week~(-1)?4 weeks)were performed to the rats.The mean arthritis index(AI)was scored every week since then.The relative concentrations of ES,IL-I?,TNF-?in sera collected at the fourth week were evaluated by western blotting. Results①The titer of the purified rAD-GFP-ES and rAD-GFP was 6.6?10~(12)pfu/ml and 4.8?10~(12)pfu/ml,re- spectively(A_(260nm)/A_(280nm)＞1.3).②The concentration of ES in sera of the group treated with rAD-GFP-ES was 2.4-lold higher compared to the normal group.③The mean arthritis index of the group treated with rAD-GFP- ES was much lower than that of the model group.The administration of rAD-GFP-ES could significantly de- creas the production of IL-1?and TNF-?in sera.Conclusions①The rAD-GFP-ES is efficiently expressed in vivo.②The rAD-GFP-ES has an inhibitory effect on the arthritis index of rat CIA.③IL-1?and TNF-?are involved in the pathogenesis of RA.The rAD-GFP-ES has an inhibitory effect on the expression of IL-1?and TNF-?in rat CIA.

Objective To evaluate the clinical application and safety of CT-guided pereutaneous transthoracic biopsy in the diagnosis of mediastinal masses.Methods Thirty three cases were undertaken CT- guided percutaneous transthoraeie biopsy with automatic biopsy gun and then the sampling specimens were undergone histological examination.The accuracy of puncture,diagnostic correctness and complications were analyzed.Results The operations were performed successfully in all 33 cases(100%),the definite pathologic diagnosis were made in 28 out of 33 cases(85%)and no complications occurred.Conclusion As for midiastinal masses,CT-guided percutaneous transthoracic biopsy is a feasible,successful,efficient interventional diagnostic method with high accuracy in localization,puncture,diagnosis and few complications, which should he recommended in clinical use more widely.(J Intervent Radiol,2007,16:852-854)

OBJECTIVETo investigate the prognostic values of HIF-1α, APE1, VEGF, and COX-2 protein expressions and their predictive value of tumor necrosis rate and prognosis in osteosarcoma, as well as their interrelationships.

METHODSFormalin-fixed paraffin-embedded tissue samples were obtained from patients with osteosarcoma. Immunohistochemical assay was performed in pre-chemotherapy samples to determine the HIF-1α, VEGF, APE1, and COX-2 protein expression levels. Hematoxylin-eosin staining was used in post-operative samples to determine the tumor necrosis rate. Univariate and multivariate analyses were used to assess the impact of protein expression on prognosis.

RESULTSTumor tissues were obtained from 49 patients. Their median follow up was 29 months. HIF-1α was significantly correlated to every protein we tested: VEGF (P = 0.032), APE1 (P < 0.001), and COX-2 (P < 0.001). HIF-1α protein expression had a significant impact on disease free survival (P = 0.006). Expression of HIF-1α had a sensitivity of 64.7% and a specificity of 71.9% for poor pathological response (< 90% of tumor necrosis) versus good pathological response to chemotherapy (≥ 90% necrosis).

CONCLUSIONExpression of HIF-1α is a predictor of tumor response to neoadjuvant chemotherapy and outcome in osteosarcoma and is correlated with VEGF, APE1, and COX-2 expression.

Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bone Neoplasms ; drug therapy ; metabolism ; pathology ; Chemotherapy, Adjuvant ; Child ; Cyclooxygenase 2 ; metabolism ; DNA-(Apurinic or Apyrimidinic Site) Lyase ; metabolism ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Osteosarcoma ; drug therapy ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; metabolism ; Young Adult

OBJECTIVETo explore the biological mechanisms underlying Angelica sindsis polysaccharide (ASP) -induced aging of human-derived leukemia stem cells (LSCs) in vitro.

METHODAcute myelogenous leukemia stem cells were isolated by magnetic activated cell sorting (MACS). The ability of LSC proliferation treated by various concentration of ASP(20-80 mg · L(-1)) in vitro for 48 hours were tested using cell counting Kit-8 ( CCK8) , colony forming were evaluated by methylcellulose CFU assay. The ultra structure changes of AML CD34+ CD38- cells were analyzed by transmission electron microscopy. The aging cells were detected with senescence-β-galactosidase Kit staining. Expression of aging-related p53, p21, p16, Rb mRNA and P16, Rb, CDK4 and Cyclin E protein were detected by quantitative reverse transcription polymerase chain reaction( qRT-PCR) and Western blotting, respectively.

RESULTThe purity of the CD34 + CD38 - cells is (91.15 ± 2.41)% after sorted and showed good morphology. The proliferation of LSC was exhibited significantly concentration-dependent inhibited after exposure to various concentration of ASP. Treated by 40 mg · L(-1) ASP for 48 hours, the percentage of positive cells stained by SA-β-Gal was dramatically increased (P < 0.01) and the colony-formed ability has been weakened (P < 0.01). The observation of ultrastructure showed that cell heterochromatin condensation and fragmentation, mitochondrial swelling, lysosomes increased in number. Aging-related p53, p21, p16, Rb and P16, Rb were up-regulated, protein regulatory cell-cycle CDK4 and Cyclin E were down-regulated. ASP may induce the senescence of LSCs effectively in vitro, P16-Rb cell signaling pathway play a significant role in this process.

CONCLUSIONASP can induce human leukemia stem cell senescence in vitro, the mechanism involved may be related to ASP regulation P16-Rb signaling pathways.

Angelica sinensis ; chemistry ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Cellular Senescence ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Gene Expression Regulation, Leukemic ; drug effects ; Humans ; Leukemia ; drug therapy ; genetics ; metabolism ; physiopathology ; Neoplastic Stem Cells ; cytology ; drug effects ; Polysaccharides ; pharmacology ; Signal Transduction ; drug effects

Leukemia is a type of malignant tumors of hematopoietic system with the abnormal increased immature leukemia cells showing metastasis and invasion ability. Liver is one of the main targets of the leukemia cells spread to, where they may continue to proliferate and differentiate and cause liver function damage, even liver failure. Our previous studies showed that Angelica polysscharides (APS), the main effective components in Angelica sinensis of Chinese traditional medicine, was able to inhibit the proliferation and induced differentiation of the leukemia cells, however, its effect on the liver during the treatment remains elucidated. In the present study, the human leukemia NOD/SCID mouse model were established by implantation human leukemia K562 cells line, then the leukemia mouse were treated with APS, Ara-c or APS + Ara-c respectively by peritoneal injection for 14 days, to explore the effect and mechanism of the chemicals on the mouse liver. Compared to the human leukemia NOD/SCID mouse model group with the treatments of APS, Ara-c and APS + Ara-c, We found that severe liver damage and pathological changes of the liver were able to alleviate: First, the number of white blood cells in the peripheral blood was significantly lower and with less transplanted K562 leukemia cells; Second, liver function damage was alleviated as liver function tests showed that alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBiL) were significantly reduced, while the albumin (Alb) was notably increased; Third, liver antioxidant ability was improved as the activities of the antioxidant enzymes glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) were significantly increased, and the contents of GSH and malonaldehyde (MDA) were decreased significantly in the liver; Fourth, the inflammation of the liver was relieved as the level of IL-1beta and IL-6, the inflammatory cytokines, were decreased significantly in the liver. Fifth, liver index was increased as the pathological observation showed that leukemia cells with diffused infiltration into the liver lobules were significantly reduced and with a remarkable increase of apoptotic positive cell rate by TUNEL test. Furthermore, the APS + Ara-c combined administration showed an even more significant positive effect. In conclusion, the APS, Ara-c therapy reduced the accumulation of leukemia cells within the liver, reduced the liver function damage and levels of inflammatory factors, improved antioxidant capacity of the liver tissue and thus alleviate the pathological changes of the liver. Moreover, the APS + Ara-c combination therapy may have an additive effect.
Angelica ; chemistry ; Animals ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cytarabine ; administration & dosage ; Humans ; K562 Cells ; Leukemia ; drug therapy ; Liver ; drug effects ; Male ; Mice ; Mice, SCID ; Polysaccharides ; administration & dosage

BACKGROUNDControl of hypersecretion of certain hormones is one of the key targets in the treatment of pituitary adenomas. RNA interference has been shown to inhibit protein expression, and thus it may represent a promising method for the treatment of pituitary adenomas. In the present study, transfection efficiency of small interfering RNA (siRNA) was optimized in human prolactinoma cells.

METHODSFirst, a method was optimized to extract highly purified human prolactinoma cells in vitro. The extracted cells were verified to retain the physiological features of prolactin (PRL) secretion. Second, three conditions for siRNA transfection were tested by the evaluation of transfection efficiency and cell viability. The proper transfection condition was verified for human prolactinoma cells. Third, the siRNA for prolactin was transfected into the human prolactinoma cells, and the suppression of PRL mRNA was evaluated by quantitative real-time reverse transcription-PCR.

RESULTSThe siRNA of 100 pmol with Lipofectamine 2000 of 5 µl for 1 × 10(6) cells was proved preferable, with transfection efficiency being 53.3% and cell viability being 69.7%. In the preliminary experiment the siRNA against PRL decreased the mRNA of PRL by 34.0%.

CONCLUSIONIt is possible to inhibit hormone hypersecretion by RNA interference, that may eventually enable therapeutic siRNA drugs developed.

Adolescent ; Adult ; Cell Line, Tumor ; Cell Separation ; Female ; Humans ; Male ; Middle Aged ; Pituitary Neoplasms ; pathology ; therapy ; Prolactinoma ; pathology ; therapy ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo compare the cell viability of chondrocytes between the anterior and the posterior spinal growth plates in adolescent idiopathic scoliosis (AIS) by proliferation and apoptosis labelling.

METHODSSeventeen AIS patients (4 male and 13 female, mean age 13.6 years old, ranged from 10 to 17 years old) were recruited in this study. Growth plates were harvested during anterior and posterior surgery. PCNA (proliferating cell nuclear antigen) and TUNEL (terminal deoxynucleotide transferase-mediated nick end labeling) were used for proliferation and apoptosis labeling on chondrocytes respectively.

RESULTSIn AIS, the distribution of the proliferating nests were denser and more parallel in anterior column than those in posterior under microscope observation. In the proliferative and hypertrophic zone the PCNA index and PCNA/TUNEL ratio were higher in the anterior column than those in the posterior column (P < 0.05 and P < 0.01 respectively). While in resting zone the differences were not so significant.

CONCLUSIONIn adolescent idiopathic scoliosis the growth viability of chondrocytes is more vigorous in anterior spinal column than in the posterior column.

Adolescent ; Apoptosis ; Cell Proliferation ; Cell Survival ; Child ; Chondrocytes ; cytology ; Female ; Growth Plate ; pathology ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Scoliosis ; pathology ; physiopathology ; Spine ; growth & development ; pathology

OBJECTIVETo compare the wear between the enamel and two types of dental decoration porcelains for all-ceramic restorations (Vita-alpha, Vintage AL).

METHODSFriction coefficients, wear scar width, element concentrations and wear surface evolution were considered relatively to the tribology of that in vivo situation. The wear scars of the samples were characterized by means of dynamic atomic force microscopy (DFM). The different element concentrations of the surface before/after the wear test were determined with energy dispersion spectrometry (EDS).

RESULTSThe friction coefficient varied from time in each kind of material. The statistical differences between materials were observed in wear scar width and properties of materials (P<0.05). DFM results showed wear surface of natural tooth full of abrasive particles and denaturation of dental texture. Wear surface of veneering ceramics consisted mainly of abrasive particles, plough and microcracking. EDS results showed that the element concentration of Fe was obviously found on the samples after wear.

CONCLUSIONThe main underlying mechanisms of natural teeth wear are abrasive, and denaturation of dental texture. Abrasive wear, adhesion and fatigue of veneering ceramics characterize the wear patterns which plays different role in Vita-alpha and Vintage AL. The wear patterns of veneering ceramics can be described as mild wear.

Ceramics ; Dental Enamel ; Dental Porcelain ; Dental Restoration Wear ; Materials Testing ; Surface Properties ; Tooth Attrition

Objective Type 1 diabetes mice model was established to investigate the changes of key enzymes involved in testosterone synthesis in testes of early diabetic mice.Methods Tatolly 20 male C57 mice were randomly divided into two groups:control and diabetic groups,and the diabetes mice were ip administered with a single dose of 150 mg/kg Streptozotocin.Four weeks after confirmation of diabetic model,the serum and testis were collected for further study.The qRT-PCR method was used to measure the expression of LHR and steroidogenesis synthetase StAR,P450scc,3β-HSD6,P450c17a1,and 17β-HSD3 mRNA.ELISA assay was performed to measure the levels of testosterone and luteinizing hormone (LH) in serum,and the enzymatic activities of 3β-HSD1,1P450c17 and 17β-HSD3 in testis tissue.Results Compared to control group,the levels of testosterone and LH of diabetic group declined significantly (P ＜ 0.05) after four weeks.The mRNA levels of LHR,StAR,CYP11a1,Hsd3b6,CYP17a1 and Hsd17b3,and enzymatic activities of 3β-HSD6,P450c17 and 17β-HSD3 were also decreased significantly compared with control group (P ＜ 0.05,0.01 and 0.001).Conclusion Expression of key enzymes of testosterone synthesis in testis of early diabetic mice decreases significantly.

Objective To investigate the mechanical prosperity and degradation rate of the scaffolds with compounding collagen and the nano sol-gel derived bioactive glass were studied,and provide the theoretical basis for the further application of collagen based scaffolds.Method The scaffold with compounding collagen and the nano sol-gel derived bioactive glass(58S)were prepared using the freeze-drying techniques with the bioactive glass as phase addition.By affecting the aggregation state of the collagen fibers with adjusting the supplementation of bioactive glass to change the microstructures of the compound scaffolds and finatly the compound scaffolds with different mechanical properties were prepared.Results (1)As the aggregation state of the collagen fibers changes,the scaffolds with the coarser collagen fibers is prepared with the diameters 400-600 nm approximately.The coarser collagen fibers will play an important role in improving the mechanical property and slowing down the degradation rate of the collagen based scaffolds.(2)The interactions between bioactive glass and collagen are studied by FTIR and Raman technologies.When the quality of content of collagen in the compound scaffolds is lower than 20%,the secondary structure of collagen is damaged severely.Conclusion The composite scaffolds with the mass ratio of collagen to bioactive glass 40:60 has the best performance in mechanical property and degradation,which will be helpful for further applications.