AIM:To detect tissue factor(TF)level both in plasma and in tissue of hepatocellular carcinoma(HCC)patients and to elucidate their association with clinical features.METHODS:Plasma TF levels of 50 cases of HCC patients and 30 cases of control were detected by ELISA.27 HCC tissue samples with their adjacent tissue samples and 27 normal liver tissues were detected by RT-PCR.RESULTS:① Plasma TF levels were increased significantly in HCC group when compared with control(P

AIM: To study the expression of laminin and survivin in primary gallbladder carcinoma(PGC),and their correlation with clinicopathologic characteristics of PGC.METHODS: Forty-nine specimens with PGC,21 with cholecystoadenoma and 13 with chronic cholecystitis were included in this study.Laminin and survivin expression in gallbladder tissues were detected by immunohistochemistry.The relationship between laminin or survivin expression and clinico-pathologic data were also analyzed.RESULTS: An intact basement membrane(BM) with well-delineated,smooth and continuous line-like laminin staining was identified in the tissues from chronic cholecystitis,adenoma of gallbladder and gallbladder carcinoma in situ.Submucosal BM,which was interrupted in PGC specimens in NevinⅡstage,was almost lost in tumor tissues in Nevin Ⅲ,Ⅳ and Ⅴ stage.Meanwhile,a distinguishing feature of the tumor tissue was broken or discontinuous line-like laminin staining in the matrix to tumor cells.These lines were detected to be weak,or even entirely absent in some neoplastic tissue sections.Sometimes,a faint cytoplasmic staining of laminin was also found in tumor tissues as classified to Nevin Ⅲ,Ⅳ and Ⅴ stage.Expression of survivin in PGC tissues was significantly higher than that in adenoma of gallbladder and chronic cholecystitis.However,survivin expressions had no specificity and positive predictive value for cell differentiation and grade as well as clinic stage of PGC by using statistical analyses.Furthermore,no correlation was confirmed between the staining features of laminin and survivin expression.CONCLUSION: Laminin expression is strongly associated with the malignancy and invasiveness of PGC,and survivin might play an important synergistic role in the development of PGC.

Objective To primarily study the possible biological significance of yon Willebrand factor (vWF) expression at the fracture site in the mechanism of fracture healing. Methods A total of 28 male SD rats were selected to set up femoral fracture models. Then, tissue samples from fracture site were randomly taken at days 2, 3, 5, 7, 10, 14 and 21 after establishment of the models, with 4 rats at each time point, vWF expression at fracture site and the relationship of vWF expression with osteoblasts and chondroblasts were observed by using computer image analysis system. The biological significance of vWF expression at fracture site in mechanism of fracture healing were analysed based on the results of vWF expression of in vitro cultured rat osteoblasts. Results The expression of vWF at every time point was positive and distributed disproportionally irregular at bone trabecula and soft tissue of the fracture site. vWF expressed positively in the extracellular matrix around the chondroblasts. Conclusions During the course of fracture healing, there may be vWF-platelet activation pathway that can regulate differentiation and proliferation of local osteoblasts and chondrocytes and activate platelets secreting growth factors such as PDGF, IGF-1, EGF and TGF-a, as creates a sound microenvironment for fracture healing.

Hypoxia is a general characteristic of most solid malignancies and intimately related to cancer progression. Homeostatic response to hypoxia is primarily mediated by hypoxia inducible factor-1alpha (HIF-1alpha) that elicits transcriptional activity through recruitment P300 coactivator. Targeting the interaction of HIF- alpha and P300 would thus constitute a novel approach for cancer treatment by suppressing tumor angiogenesis and metastasis. Here, a screening assay was developed for inhibitors targeting the interaction between HIF-1alpha and P300. The nucleotide sequence of human HIF-1alpha and P300 were cloned into pBIND and pACT vectors, named pBIND-HIF1alpha and pACT-P300. The interaction of HIF-1alpha and P300 was identified in HEK293 cell using mammalian two-hybrid system. And compound chetomin decreased their interaction in this mammalian two-hybrid system. We further verified HIF-1 inhibition effect of chetomin in U251-HRE cells. Therefore, we established a screening assay combined HIF-1alpha and P300 mammalian two-hybrid system and U251-HRE reporter assay for HIF-1 selective inhibitors.
Cell Hypoxia ; Disulfides ; pharmacology ; Drug Screening Assays, Antitumor ; E1A-Associated p300 Protein ; antagonists & inhibitors ; HEK293 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; antagonists & inhibitors ; Indole Alkaloids ; pharmacology ; Two-Hybrid System Techniques

The aim of this study is to evaluate anti-tumor activities and mechanism of a novel kinase inhibitor ZLJ213 which targeted Aurora A and vascular endothelial growth factor receptor (VEGFR) in vitro and in vivo against human colon cancer. Results showed that ZLJ213 inhibited cell proliferation and induced cell cycle arrest and apoptosis of HCT1 16 and SW48 cell lines. In HCT116-derived xenograft, ZLJ213 dosed at 100 mg · kg(-1) inhibited tumor growth by 73.24%. The IC50 of ZLJ213 on the expression of p-Aurora A was 0.258 µmol · L(-1) analyzed by ELISA. Under the concentration of 0.08 µmol · L(-1), ZLJ213 could inhibit the activities of Aurora A, Histone H3 and VEGFR of HCT116 and SW48 cell lines. Simultaneously, ZLJ213 induced activation of Caspase 3 and PARP cleavage. Above data suggested that ZLJ213 had the ability to inhibit cell proliferation and induce cell apoptosis both in vitro and in vivo in colon cancer, and down-regulate the expression of p-Aurora A and p-VEGFR. ZLJ213 might be a potential therapeutic agent against colon cancer.
Animals ; Apoptosis ; Aurora Kinase A ; antagonists & inhibitors ; Cell Cycle Checkpoints ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Colonic Neoplasms ; pathology ; Humans ; Protein Kinase Inhibitors ; pharmacology ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; Xenograft Model Antitumor Assays

BACKGROUND:The intracel ular accumulation of reactive oxygen species leads to oxidative stress. Hypoxia is widespread in physiological and pathological condition. Variation of bone proliferation and differentiation when bone tissues cultured or bone cel s induced toxicity by reactive oxygen species under hypoxia have not yet been reported. OBJECTIVE:To observe the biological characteristics of MC3T3-E1 pretreated with different concentrations of hydrogen peroxide (H2O2) in hypoxia, thus understanding the cel mechanism underlying prolonged bone healing in the elderly with osteoporosis and diabetes. METHODS:The MC3T3-E1 cel s pretreated with different concentrations of H2O2 were cultured in different oxygen concentrations. The proliferation of MC3T3-E1 was detected by cel counting kit-8. The cel differentiation was detected through alkaline phosphatase staining and alizarin red staining. Total RNAs were extracted and used for analyzing the mRNA levels of col age type 1, alkaline phosphatase and Cbfa1. RESULTS AND CONCLUSION:When MC3T3-E1 pretreated with 200μmol/L H2O2 for 6 hours, the cel proliferation was increased with time, but lower than that in the control group. The alkaline phosphatase activity was weakened, and the number of mineralized nodes was decreased at the early stage of differentiation. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours, the cel proliferation was decreased obviously. The alkaline phosphatase activity was stil weakened, and the number of mineralized nodes was decreased further, but not affected by hypoxia. When MC3T3-E1 pretreated with 400μmol/L H2O2 for 6 hours and then cultured in hypoxia, the mRNA expression of Cbfa1 was decreased, but the mRNA expressions of col age type 1 and alkaline phosphatase were significantly increased. These results suggest that MC3T3-E1 pretreated with low concentration of H2O2 show a significant decrease in proliferation, while MC3T3-E1 pretreated with a high concentration of H2O2 and cultured in hypoxia show a decrease in osteogenic differentiation, especial y at the early stage of alkaline phosphatase formation.

Objective To make sciatic nerve traction injury models of rabbit , in order to prospectively evaluate possibility and accuracy of diffusion tensor tracking (DTT) in sciatic nerve injures. Methods The right sciatic nerves of 32 New Zealand white rabbits were selected for traction injury , and the left sciatic nerves were the sham operation side. DTI scan was performed before and after the operation on 1st day , 3rd day, and 1 week, 2, 3, 4, 6 and 8th week respectively, and DTT was reconstructed. Then the length of reconstructed fiber tracts and fiber density index were calculated. After the MRI scan , the sciatic nerve was removed to perform pathologic examination at different time points. Results The difference of the length of reconstructed fiber tracts between nerve traction injury and sham operation side was significant at 1 day~2weeks after operation (P<0.05), while the difference was not significant at 3~8 weeks.The fiber density index of nerve traction injury and sham operation side was significantly different at 1 day~8 weeks(P<0.05). 1day after operation, myelin sheath of traction portion was obviously twisting. 3 days after operation , a large amount of myelin sheath broke down. 2 weeks after operation , axon, myelin sheath degeneration and regeneration coexisted at the same time. 8 weeks after operation , nerve fibers regenerated and restored to normal structure. Conclusion The length time curve and density index-time curve of nerve traction injury are consistent with the changes of pathology , which can be used as a supplementary method to evaluate the degeneration and regeneration in nerve injury.

Objective: To study the protective effect of Xiaoyao Tablet(XYT)(Radix Bupleuri, Radix Angelicae Sinensis, Rhizoma Atractylodis Macrocephalae, Radix Paeoniae Alba, Poria, Radix Glycyrrhizae, Herba Menthae, Roasted ginger) on liver. Methods: Using the acute hepatic injury model induced by Tetrachloride and D-galactosamine (CD-GaIN) in rats or mice, we observed the influence of XYT on ALT and AST level in the serum, MDA level and glutathione-s-transferase(GST) activity in the liver homogenates. The pain-killing effect was tested by body twisting model induced by glacial acetic acid and by hot plate model in mice. Results: The XYT reduced the serum ALT level significantly in the acute hepatic injury model in rats and mice. In the CCl 4 reduced acute hepatic injury mice model, XYT decreased the MDA level and increased the GST activity in liver significantly, also it reduced the MDA level in the serum. The XYT reduced twisting number in mice induced by glacial acetic acid. In the hot plate test, the pain threshold was increased. Conclusion: The drug has the similar hepatoprotective function to Xiaoyao Pills.

Objective To investigate the correlation between eigenvalues(λ∥ andλ⊥)of sciatic nerve and limb function in a rabbit model of acute nerve traction inj ury for finding the role ofλ∥ andλ⊥ on diagnosis of nerve inj uries.Methods The right sciatic nerves of 32 Newland rabbits were selected to be performed as traction injury,and then left sciatic nerves were treated as control for the sham operation side.MRI DTI scan was performed before and after the operation on 1 day ,3 day,and 1 week,2,3,4,6 and 8 weeks respectively.Theλ∥ andλ⊥ of inj ured sciatic nerves and sham operated nerves were measured,meanwhile functional exami-nations were evaluated,and then analyzing the correlation betweenλ⊥ of inj ured sciatic nerves and the score of inj ured limb function in the periods of 3 days to 8 weeks;finally different portions of sciatic nerve were removed for histological examinations.Results Theλ⊥ of inj ured sciatic nerves in the proximal portion was slightly increased at 3 days and recovered to the normal at 2 weeks.Theλ⊥ of inj ured sciatic nerves in the traction portion and distal portion were slightly raised at 1 day,reached to its maximum value at 3 days,and then decreased gradually after 1 week and dropped to the pre-operation level after 4 weeks.Theλ⊥ of inj ured sciatic nerves in the proximal portions was significantly higher than that of the sham-operated nerves from 1 day to 1 week(F=7.275,P=0.000). Theλ⊥ of inj ured sciatic nerves in the traction portion and distal portion were significantly higher than that of the sham-operated nerves from 1 day to 3 weeks(F=5.851,F=3.794,P=0.001,P=0.000).However,there was no significantly difference between theλ∥ of inj ured nerves in each portion and sham-operated nerves .The negative correlation between theλ⊥ of inj ured sciatic nerves in the distal portion and the score of the nerve function in the injured limb was found (r=-0.938,r=-0.897,P<0.01).Conclu-sion Theλ⊥ of inj ured nerves in the traction portion and distal portion could monitor the process of degeneration and regeneration of sciatic nerve in traction inj ury,while theλ∥ plays no obvious role in diagnosis of nerve inj uries.

The efficacy of constructivism teaching mode in physical diagnostics teaching was evaluated in this study. We built up the constructivism teaching mode in diagnostics teaching taking the clinical symptoms as the theme, and through such aspects as courseware design, teaching plan preparation, SP playing and inquiry, SimMan simulated physical examination, condition analysis, etc. Then questionnaires were conducted to analyze the role of the constructivism teaching mode in diagnostics teaching. The diagnostics constructivism teaching mode can provide students with a platform for self-construction of diagnostics and integrated application of knowledge. Meanwhile, students' sense of participation can be improved and multiple learning skills are enhanced during the course.