Aim To investigate the effect and mechanism of valdecoxib on the apoptosis of human esophageal cancer cells.Methods Flow cytometry was used to observe the effect of valdecoxib on apoptosis and the cell cycle distribution of Eca109 cells.Transmission electron microscope was further used to detect the cell apoptosis.The content of LDH was examined using LDH kit.The expressions of p-p38MAPK,Fas and FasL protein were detected using flow cytometry.Results Valdecoxib of 25～400 ?mol?L~(-1) significantly induced the apoptosis of Eca109 cell line,and the rate of apoptosis was increased from(2.95?0.83)% to(48.46?0.73)%,50～400 ?mol?L~(-1) valdecoxib also decreased the proliferation index and the proportion of cells in the S phase,increased the proportion of cells in the G_0/G_1 phase,but had no effect on the proportion of cells in the G_2/M phase.Compared with those in Eca109 cells cultured in the medium with solvent,the expression of p-p38MAPK,Fas and FasL was higher in the Eca109 cells exposed to valdecoxib in a dose-dependent manner in 72 h.Conclusion Valdecoxib can induce apoptosis of Eca109 cell line partly by up-regulating the expression of p-p38MAPK/Fas/FasL.

Objective To establish the carotid fusiform aneurysm model in rabbits carrying similar characteristics of human intracranial aneurysms by using induction method with porcine pancreatic elastase. Methods Twenty-five New Zealand white rabbits were randomly divided into normal control group (n=5), saline control group (n = 5) and study group (n = 15). The rabbits of the study group were randomly and equally subdivided into 7-day subgroup, 14-day subgroup and 21-day subgroup. By using induction method with porcine pancreatic elastase to digest right common carotid the fusiform aneurysm model was established in all the rabbits of the study group. DSA examination , HE staining and elastic fiber staining pathologic examination were carried out at 7, 14 and 21 days after the procedure to observe the imaging and pathologic changes of the fusiform aneurysm models. Results DSA angiography showed that the mean vascular diameters of the normal control group and the saline control group were (1.64 ± 0.17) mm and (1.66 ± 0.24) mm respectively. The mean length and width of the fusiform aneurysm of the 7-day subgroup, 14-day subgroup and 21-day subgroup were (19.33 ± 1.65) mm and (2.86 ± 0.21) mm, (19.66 ± 1.18) mm and (3.95 ± 0.54) mm, and (19.84 ± 0.82) mm and (4.03 ± 0.95) mm, respectively. Pathologically, rupture of internal elastic membrane, disordered structure of tunica media smooth muscle and distortion of cell shape were observed in the rabbits of 7-day subgroup. Gradually stabilized aneurysmal lumen intimal hyperplasia was seen in the rabbits of 14-day subgroup. Remarkable structure changes at the aneurysmal neck-cavity junction were found in the rabbits of 21-day subgroup. Elastic fiber staining demonstrated that strikingly thinned elastic layer was observed in the rabbits of 7-day subgroup, gradually thinning elastic layer at the aneurysmal neck-cavity junction was seen in the rabbits of 14-day subgroup, and the thinned elastic layer became stable in the rabbits of 21-day subgroup. Conclusion Using simple surgical method combined with porcine pancreatic elastase to digest vascular wall, carotid fusiform aneurysm models can be reliably established in New Zealand white rabbits which carry similar morphologic and pathologic characteristics of human intracranial aneurysms.

BACKGROUND:Vascular endothelial growth factor (VEGF) is a crucial factor for regulating bone marrow mesenchymal stem cells in microenvironment, which can promote endothelial differentiation of bone marrow mesenchymal stem cells. But there is no report about VEGF effect on regulating the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. OBJECTIVE:To investigate the regulatory role of VEGF in the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:The bone marrow mesenchymal stem cells were isolated and cultured from mouse long bones. cellCounting Kit-8 was used to test the proliferation of bone marrow mesenchymal stem cells cultured with different concentrations of recombinant VEGF proteins. The appropriate concentration of VEGF recombinant protein was selected to test their effects on the osteogenic differentiation of bone marrow mesenchymal stem cells. The mRNA and protein levels of Osterix, Runx2, alkaline phosphatase, osteocalcin and heme oxygenase-1 in bone marrow mesenchymal stem cells under induction of VEGF were detected by molecular biology methods. RESULTS AND CONCLUSION:(1) VEGF promoted the proliferation of bone marrow mesenchymal stem cells dose-dependently, and 100μg/L was the optimum concentration. (2) VEGF promoted the expression of Osterix, Runx2, alkaline phosphatase and osteocalcin in bone marrow mesenchymal stem cells under osteogenic induction. Similar results were obtained by alizarin red staining and quantification of numbers of calcium nodules. (3) VEGF induced the expression of heme oxygenase-1 in bone marrow mesenchymal stem cells at mRNA and protein levels. These findings indicate that VEGF can induce the expression of heme oxygenase-1 i in bone marrow mesenchymal stem cells, and promote the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells.

Objective To investigate the effect and possible mechanisms of high mobility group box (HMGB) 1 on the proliferation of RSC-364 synoviocytes. Methods ① RSC-364 cells stimulated by 10 μg/L TNF-α and cells of the normal control groups were collected at 6, 12, 24 h respectively in vitro. HMGB1mRNA and protein was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunocytochemistry (ICC); ②RSC-364 cells induced by 10 μg/L HMGB1 were collected in 6, 12, 24 h respectively, so did normal control group cells in vitro. The expression of signal transducer and activator of transcription (STAT) mRNA 1 was detected by RT-PCR. The expression of STATland SOCSI proteins were detected by ICC and flow cytometry analysis (FCM). The expression of PCNA was detected by ICC. Results ① Compared with the control group, TNF-α markedly up-regulated HMGBI mRNA at 6, 12, 24 h respectively [0.86, 0.92, 1.06 vs 0.70, P<0.01 ], as well as protein expression level. Positive signal of HMGB1 proteins was not only expressed in nuclear but also in cytoplasm after stimulation. ② Compared with normal group, HMGBI increased the expression of P-STAT1 mRNA and protein at 6, 12, 24 h respectively [0.30, 0.69, 1.05 vs 0.24, P<0.01 ] and [1.34±0.09，1.55±0.16，1.74±0.13 vs 1.00±0.15，P<0.01]. The expression of SOCSI protein increased significantly in HMGB1 group at 6 and 12 hours ( 1.43±0.10 vs 1.58±0.05), but it decreased at 24 hours (1.24±0.15). ③The expression of p-STATI protein was negatively correlated with that of SOCS1 protein. Conclusion HMGB1 appears to be an important mediator in the proliferation of RSC-364 cells, partly by up-regulating the expression and aetivity of p-STAT1.

Objective To investigate the effect and possible mechanism of arsenic trioxid(As2O3)on proliferation of RSC-364 synoviocyte lines stimulated with TNF-?.Methods RSC-364 synoviocytes were cultured with standard medium as control group or medium supplemented with 10 ?g/LTNF-? and different concentrations of As2O3 respectively. MTT assay were carried out to study cell proliferation. Proliferation index (PI) and cell cycle were detected by flow cytometry (FCM). RT-PCR was used to detect the mRNA expression of High mobility group box chromosomal protein (HMGB)1. HMGB-1 and proliferation cell nuclear antigen (PCNA) proteins were detected by immunocytochemistry and FCM. Results (1)As2O3 inhibited proliferation of cell lines stimulated by TNF-? time-dependently and dose-dependently. (2)Compared with normal group, TNF-? up-regulated HMGB-1 protein and mRNA as well as PCNA protein. HMGB-1 protein was not only in nuclear but also in cytoplasm by immunocy-tochemistry. As2O3 down-regulated mRNA and protein of HMGB-1 in a dose-dependent manner; so did PCNA proteins (P

Objective:To analyze the factors that affect patient prognosis after radical nephrectomy.Meth-ods:A total of 389 cases of renal cell carcinoma treated with radical nephrectomy between January 1 993 and December 2006 were reviewed.All the data were encoded.inserted into an Excel database and then ana-lyzed by SPSS 1 3.0 software.The cumulative survival rates were calculated by life-table method.We as-sessed the impact of multiple covariates on survival time with the Cox Regression model.Results:The patho-logical results showed that 307 cases were clear call carcinoma,51 cases were papillary renal cell carcinoma,21 cases were chromophobic renal cell carcinoma,2 cases were collecting duct carcinoma.and 8 cases were unclassified.One hundred and ninety-eight cases were of T1N0M0, 113 cases were of T2N0M0, 3 cases were of T1N1M0,10 cases were of T2N1M0, 51 cases were of T3N0M0, and 14 cases were of T3N1M0, Two hundred and sixty-eight cases were followed up.The 1-year survival rate was 96.5%,the 3-year survival rate was 90.7%.the 5-year survival rate was 75.7%.and the 10-year survival rate was 65.8%.Multivariable analysis revealed that significant prognostic factors included TNM stage,Robson stage.vena cava and supplementary treat-ment(X2=22.50.P=0.001).The most important prognostic factor was pathological stage(TNM and Robson).The regression coefficients were 0.533 and 0.674,and the relative risk was 1.941 and 2.01 1(P=0.004 and p=0.002).Conclusion:Radical nephrectomy is safe and effective.TNM stage.Robson stage and vena cava are prognostic factors.Supplementary treatment is a protective factor.

Objective To analyze histomorphometrical characteristics of bone and bone marrow tissue in the vertebral lamina of patients with osteofluorosis, and to explore the influencing factors on signal intensity in MRI. Methods Spinal MRI of 109 patients (57 men, 52 women;age range 32-80 years;mean age 52 years) with osteofluorosis from December 2001 to May 2012 was analyzed retrospectively, including 48 patients in cervical segment, 31 in thoracic segment and 30 in lumbar segment. 36 pa?tients (16 men, 20 women;mean age 51 years;age range 34-68 years) had undergone laminectomy and the vertebral lamina speci?mens were collected. The cervical MRI of 48 patients with matching gender and age (26 men, 22 women;mean age 51 years, age range 34-71 years) was selected as control group, who were from areas where fluorosis is not endemic. All patients were divided in?to vertebra low, medium and high signal groups according to T1WI of MRI. The vertebra signal to noise ratio measure and stan?dardization of signal intensity were performed. Osteosclerosis, osteoporosis and normal bone were differentiated under spinal X?ray plain film. Combined with histomorphometric analysis of vertebra lamina in 36 patients, correlation between MRI signal intensity, histomorphometric parameters of the vertebra lamina and influencing factors on signal intensity were studied. Results 77 pa?tients (70.6%, 77/109) had osteosclerosis indicated by appearance of spine under X?ray, 29 (26.6%, 29/109) osteoporosis and 3 (2.8%, 3/109) normal bone. T1WI of MRI showed 25 cases had low signal vertebra, 52 medium signal and 32 high signal. The ver? tebra SNR in patients with osteofluorosis was lower on T1WI, T2WI and short time inversion recovery (STIR) sequences, compared with control group. Those with a low versus high signal on T1WI had 6.04 times the odds of osteosclerosis (OR=6.04, 95%CI 2.44-14.91, P<0.001). Histomorphometry of vertebral lamina in 36 patients with osteofluorosis was performed, revealing that not only the trabecular bone volume had changed, but also did the adipocyte volume and hemopoietic cell volume in the bone marrow tis?sues. Compared with normal reference values, trabecular bone volume was significantly increased (47.7%± 13.3% vs. 14.7%± 4.3%) (P<0.001);adipocyte volume was significantly decreased (12.3%±9.1%vs. 50.5%±8.7%);hematopoietic cell volume was decreased (40.0%±7.0%vs. 42.5%±8.5%) (P=0.038). There were inverse associations between trabecular bone volume and adipo?cyte volume (r=-0.869, P<0.001), and between trabecular bone volume and T1WI (r=-0.851, P<0.001) found by Pearson correla?tion test. In contrast, there were positive associations between T1WI and adipocyte volume (r=0.927, P<0.001). Conclusion The vertebra T1WI signal intensity is decreased in patients with osteofluorosis, resulting from increase of trabecular bone volume and re?duction of adipocyte volume. The vertebra STIR signal intensity is decreased, mainly caused by increase of trabecular bone volume.

OBJECTIVETo investigate the probabilities of core-biding factor a1 (Cbfa1) expression by human skin fibroblasts induced in vitro.

METHODSThe fibroblasts were isolated, purified from human skin, and were grown in incubation in the media of TNF-alpha, BMP-2, and combined TNF-alpha and BMP-2 at certain concentrations, respectively. The changes in biological features of these fibroblasts correlated with osteogenesis were detected by immunohistochemistry and RT-PCR assay.

RESULTSTNF-alpha could switch phenotype of collagen in fibroblasts from Type I and III to Type I and induce fibroblasts to express Ras and BMP type I receptor (BMPR-IA). TNF-alpha in combination with BMP-2 could induce fibroblasts to express Cbfa1 and osteocalcin mRNA.

CONCLUSIONHuman skin fibroblast could be induced into pro-osteoblast expressing Cbfa1, an osteoblast-specific transcription factor and a regulation of osteoblast differentiation, and combined use of TNF-alpha and BMP-2 was one of the regulating factors.

Bone Morphogenetic Protein 2 ; Bone Morphogenetic Protein Receptors, Type I ; Bone Morphogenetic Proteins ; pharmacology ; Cells, Cultured ; Collagen ; biosynthesis ; Core Binding Factor Alpha 1 Subunit ; Core Binding Factors ; Fibroblasts ; metabolism ; Humans ; Neoplasm Proteins ; Osteocalcin ; biosynthesis ; Protein-Serine-Threonine Kinases ; biosynthesis ; RNA, Messenger ; analysis ; Receptors, Growth Factor ; biosynthesis ; Skin ; cytology ; Transcription Factors ; biosynthesis ; genetics ; Transforming Growth Factor beta ; Tumor Necrosis Factor-alpha ; pharmacology

Aim To investigate the regulatory effect of p38MAPK signal pathway on the apoptosis of human esophageal cancer cells induced by valdecoxib.Methods The tumor cells were inoculated in the dose of 1?107?L-1.After 6 h,the cells were divided into control group,solution group,400,200,100,50,25 ?mol?L-1 valdecoxib group and various concentration valdecoxib+SB203580 group,cultured for 72 h.FCM and DNA Ladder were used to detect the apoptosis of Eca109 cells.p38 mRNA expression was detected by RT-PCR.The expression of p-p38MAPK protein was detected by immunohistochemical staining and FCM.Results ① Valdecoxib could increased the apoptosis rate of Eca109 cell in concentration-dependent fashion.Apoptosis rate was increased to 48.46% in 400 ?mol?L-1 valdecoxib group at the incubation time of 72 h.DNA ladder was the most recognized marker of apoptosis,and there was obvious DNA ladder in valdecoxib treated group,especially in 400 ?mol?L-1 group.② Valdecoxib could increase the expression of p38MAPK,while SB203580 could inhibit the over-expression induced by valdecoxib,at the same time,the apoptosis rate was been decreased.③ The expression of p38MAPK protein was positively correlated with the apoptotic rate(r=0.822,P=0.000).Conclusions Valdecoxib could activate p38MAPK pathway,thus induce the apoptosis of Eca109 cells,which may be one of the mechanisms for the inhibition of cell growth by valdecoxib.