Clinical significance of HBME-1,Galectin-3,and CK19 expression and the status of BRAF mutation in papillary thyroid carcinoma

Objective The aim of this study was to explore the clinical significance of the expression of proteins human bone marrow endothelial cell markers(HBME-1), Galectin-3, and cytokeratin19(CK19), as well as the status of v-raf murine sarcoma viral oncogene homolog B1(BRAF) mutation in papillary thyroid carcinoma(PTC). Methods Immunohistochemical staining was performed in 82 specimens each of PTC and papillary benign lesions to detect the expression of HBME-1, Galectin-3, and CK19. Polymerase chain reaction(PCR) and gene sequencing were performed on 60 specimens each of PTC and papillary benign lesions to detect the status of BRAF mutation. Results The positive expression ratios of HBME-1, Galectin-3, and CK19 in PTC were 98.8%, 97.6% and 100% respectively, which were significantly higher than the expressions in papillary benign lesions(P < 0.05). No significant relationship was observed between the expression of these makers and the clinicopathological features of PTC. The sensitivity of co-expression of HBME-1 and CK19 or HBME-1 and Galectin-3 as diagnostic criteria of PTC was 99.9%, with a specificity of 95.4%. BRAF mutation was detected in 40 of 60 PTC(66.7%) specimens. There was a statistical difference in BRAF mutations between PTC and papillary benign lesions(P < 0.05); there were no associations between BRAF mutation and the clinicopathological features of PTC. Conclusion Combined immunohistochemical staining of HBME-1, Galectin-3, and CK19 can further improve the sensitivity and specificity of differential diagnosis of PTC. BRAF mutation is a significant genetic event, which may have diagnostic value for PTC.

Endostatin promotes the anabolic program of rabbit chondrocyte

Endostatin is a natural occurred angiogenesis inhibitor derived from collagenXVIII. So far its function during the angiogenesis process of bone formation and arthropathy has not been well studied yet. The present study addresses the function of endostatin in rabbit articular chondrocytes (RAC). We found that endostatin can promote RAC adhesion and spreading as well as its proliferation. In monolayer cultured RAC, CollagenII, TIMP1 and collagenXVIII transcription were up regulated by endostatin while collagenI and MMP9 were down regulated. Moreover collagenXVIII and endostatin antigens are present at synovial fluid. These findings indicate new function of endostatin as a homeostatic factor in cartilage metabolism.

EFFECTS OF TGF β1 AUTOCRINE BLOCKAGE ONOSTEOSARCOMA CELLS

Objective To evaluate the effects of transforming growth factor β1 (TGF β1) autocrine blockage on proliferation activity and drug sensitivity of osteosarcoma. Methods Northern blot, MTT determination, and 3H thymidine incorporation were used to investigate the effects of antisense TGF β1 gene on osteosarcoma. Results The proliferation of osteosarcoma cells transfected by antisense TGF β1 gene was suppressed markedly, and adriamycin sensitivity was significantly increased. Conclusion Blockage of osteosarcoma cells TGF β1 autocrine loop inhibits cell proliferation and enhances chemother-apy sensitivity.

Intraoperative endoscopy in obstructive hypopharyngeal carcinoma

AIM： To demonstrate the necessity of intraoperative endoscopy in the diagnosis of secondary primary tumors of the upper digestive tract in patients with obstructive hypopharyngeal carcinoma. METHODS： Thirty-one patients with hypopharyngeal squamous cell carcinoma had been operated, with radical intent, at our Institution in the period between 1978 and 2004. Due to obstructive tumor mass, in 7 （22.6%） patients, preoperative endoscopic evaluation of the esophagus and stomach could not be performed. In those patients, intraoperative endoscopy, made through an incision in the cervical esophagus, was standard diagnostic method for examination of the esophagus and stomach. RESULTS： We found synchronous foregut carcinomas in 3 patients （9.7%）. In two patients, synchronous carcinomas had been detected during preoperative endoscopic evaluation, and in one （with obstructive carcinoma） using intraoperative endoscopy. In this case, preoperative barium swallow and CT scan did not reveal the existence of second primary tumor within esophagus, despite the fact that small, but T2 carcinoma, was present. CONCLUSION： It is reasonable to use intraoperative endoscopy as a selective screening test in patients with obstructive hypopharyngeal carcinoma.

Expression levels of CXCR4 and VEGF correlate with blood-borne metastatic progression and outcome in patients with osteosarcoma

Objective:We determine whether chemokine receptor CXCR4 and vascular endothelial growth factor(VEGF) expression related to the metastasis and survival outcome of patients with osteosarcoma.Methods:Tissue microarray(TMA) was used to detect the expression of CXCR4 and VEGF in 56 osteosarcoma patient samples.Two-year follow-up was performed to observe the metastatic behavior and overall survival of osteosarcoma patients.Results:There was a significant correlation between the expression levels of CXCR4 and VEGF in 56 osteosarcoma patient samples(P = 0.002).Univariate analysis revealed the expression of CXCR4 and VEGF was not associated with age, gender and the level of ALP but associated with clinical stage.Conclusion:These data raises the possibility that VEGF could regulate the levels of CXCR4 to promote the migration of tumor cells to target organs.CXCR4 and VEGF expression are highly correlated with metastatic progression in patients with osteosarcoma and their immunohistochemical expression have predictive value for the metastatic development.

Bone marrow-originated endothelial progenitor cells contribute to neovasculature of tumor

Objective: Neovascularization of tumor is a complex process. In this study, we aimed to reveal whether the bone marrow-originated endothelial progenitor cells (EPCs) contributed to neovasculature in tumor and the angiogenesis-associ-ated factors, VEGF and B-FGF, enhanced this process. Methods: We had established a mouse model, which were deprived of bone marrow by radiation and transplanted with bone marrow of syngenetic GFP (Green Fluorescence Protein)-transgened mice, then implanted Lewis cells. Immunohistochemical and immunoflourensence proved the EPCs location in tumors by indentifying colocalization of GFP expression in cells staining with endothelial progenitor cell markers, CD 133, ICAM-1, CD31. The growth statue and MVD of tumor was observed after injection of VEGF or B-FGF. ICAM-1 and VE-cadherin in tumor were detected by Western blot. Results: By immunohistochemical and immunoflourensence, we proved part of bone marrow precursors located in area of tumor angiogenesis and VEGF or B-FGF increased the MVD of tumor. In Western blot, it was found and VEGF or B-FGF up-regulate the expression of ICAM-1, VE-Cadherin. Conclusion: Bone marrow-derived endothelial progenitor cell seem to be recruited in neovasculature induced by tumor. VEGF and B-FGF are key regulators of this process.

Application of modified polyethylene glycol hydrogels in the construction of tissue engineered heart valve

To enhance the adhesion of seeding-cells to the biomaterial scaffolds, the PEG-hydrogels were modified. Porcine aortic valves were decellularized with Triton X-100 and trypsin. The cells were encapsulated into the PEG-hydrogels to complete the process of the cells attaching to the acellular porcine aortic valves. Herein, the autologous mesenchymal stem cells （MSCs） of goats were selected as the seeding-cells and the tendency of MSCs toward differentiation was observed when the single semilunar TEHV had been implanted into their abdominal aortas. Furthermore, VEGF, TGF-β1, and the cell adhesive peptide motif RGD were incorporated. Light and electron microscopy observations were performed. Analysis of modified PEG-hydrogels TEHV＇s （PEG-TEHV） tensile strength, and the ratio of reendothelial and mural thrombosis revealed much better improvement than the naked acellular porcine aortic valve （NAPAV）. The data illustrated the critical importance of MSC differentiation into endothelial and myofibroblast for remodeling into native tissue. Our results indicate that it is feasible to reconstruct TEHV efficiently by combining modified PEG-hydrogels with acellular biomaterial scaffold andautologous MSCs cells.

Effects of Buyang Huanwu Tang Combined with Bone Marrow Mesenchymal Stem Cell Transplantation on the Expression of VEGF and Ki-67 in the Brain Tissue of the Cerebral Ischemia-Reperfusion Model Rat

Objective:To explore the mechanism of Buyang Huanwu Tang (补阳还五汤 Decoction Invigorating Yang for Recuperation) combined with bone marrow mesenchymal stem cells (MSCs) transplantation in protecting nerves of cerebral ischemic injury. Methods: Local cerebral ischemia-reperfusion rat model was established with modified Zea-Longa thread-occlusion method, and MSCs were injected into the caudal vein, and Buyang Huanwu Tang(补阳还五汤)was administrated. Vascular endothelial growth factor (VEGF) and Ki-67 expression in the ischemic side of the brain in the cerebral ischemic-reperfusion rat were detected with immuno-histochemical staining method. Results: VEGF and Ki-67 expressions were significantly up-regulated in the MSCs group and the combination group, with significant differences as compared with the model group and the sham operation group (P<0.05), and with the most strongest effect in the combination group. Conclusion: Buyang Huanwu Tang(补阳还五汤)combined with MSCs transplantation repairs the injured blood vessels and lesion tissues possibly by up-regulation of VEGF and Ki-67 expression.

Combinatorial effects of NaomaiYihao Capsules(脑脉一号胶囊) and vascular endothelial growth factor gene-transfected bone marrow mesenchymal stem cells on angiogenesis in cerebral ischemic tis sues in rats

OBJECTIVE:To investigate the combinatorial effects of Naomai Yihao(NMYH) Capsules(脑脉一号胶囊) and vascular endothelial growth factor(VEGF) gene-transfected bone marrow mesenchymal stem cells(BMSCs) on angiogenesis in cerebral ischemic tissues in rats and the mechanism.METHOD:BMSCs were isolated and cultured from bone marrow by an adherence method.Then,BMSCs were transfected with the eukaryotic expression plasmid pEGFP-VEGF 165 by positive ionic liposome transfection.A rat model of middle cerebral artery occlusion(MCAO) was established.Rats were allocated to six groups:model,BMSC,VEGF gene-transfected BMSC transplantation(BMSC/VEGF),NMYH,combined NMYH and BMSC/VEGF(combined treatment group) and sham operation groups.The behavioral rating score(BRS) of rat and the expression of CD34 and VEGF in brain tis sue were measured by immunohistochemistry on days 7,14 and 21 after reperfusion.Angiogenesi was observed and evaluated with laser scanning confocal microscopy.RESULTS:The BRS of rats in NMYH,BMSC transplan tation and combined treatment groups was significantly lower than that of the model group(P< 0.001),with no significant difference between NMYH and transplantation groups(P=0.619).The expression of CD34 andVEGF in NMYH,transplanta tion and combined treatment groups increased(P< 0.001),with a significant difference between NMYH and transplantation groups(P<0.001).The blood vessel area in NMYH,transplantation and com bined treatment groups was significantly increased(P<0.05),without a significant difference between NMYH and transplantation groups(P=0.873).CONCLUSIONS:VEGF gene-transfected BMSCs im prove angiogenesis in the cerebral ischemic area NMYH Capsules promote angiogenesis in MCAO rats treated with BMSC transplantation,which show an improved BRS.The mechanism of angio genesis may be related to up-regulation ofVEGF ex pression.