Objective To evaluate the effect of tissue factor (TF), tissue factor pathway inhibitor-1,2 (TFPI-1,2) on cancer metastases and thrombosis complicated with cancer. Methods Blood samples from 292 cancer patients were collected and were divided into different teams according to cancer types,complicated with or without thrombosis; TF, TFPI-1, TFPI-2, plasma concentrations were measured by ELISA;Tissue expression of TF, TFPI-1, TFPI-2 were observed by immunohistochemical method. Results Plasma concentrations of TF and TFPI-1 in all kinds of cancers were higher than the control and lung cancer was the highest; TFPI-2 plasma concentrations had no statistics differences among all these teams. Tissue expression of TF of all kinds of cancer were higher than the adjacent tissues, lung cancer was higher than the other types of cancer. There were no statistics differences for TFPI-1 and TFPI-2. Both TF and TFPI-1 plasma concentrations of cancer with-or without-thrombosis were higher than control. TF was even higher in cancer with thrombosis team, TFPI-1 had no statistic difference between these two teams. TFPI-2 concentrations had no differences among all these teams. Conclusion Many kinds of tumor have higher expression of TF, it is expressed with different intensity according to different types of cancer. TFPI-1 has no clear effect in cancer growing and metastases. Unbalance of TF and TFPI-1 in plasma may relate to high coagulation state of cancer and may accelerate the thrombosis formation in cancer.

Objective To improve the establishment of rabbit model with VX2 liver tumor and assess sonographical value in monitoring the tumor growth.Methods Fifteen New Zealand white rabbits were implanted with VX2 tumor in two liver lobes under ultrasound guidance percutaneously.Ultrasound examinations were performed at twenty days and thirty-six days after implantation.At the same time exploratory laparotomy was performed.Results Among 30 implanted tumors of 15 rabbits,16(53%) tumors were successfully implanted with few complications.It took 15 to 40 minutes to implant tumors in one rabbit.Implantation rates of left lobe and right lobe were 53% and 50% respectively,and there was no significant difference.Ultrasound examination could monitor tumor growth well.Conclusions After the improvement,the establishment of model was easier,less time-consuming,minimally invasive and caused few complications than other implantation methods.Serial ultrasound examinations should be performed in monitoring the tumor growth.

Objective To study the relationship between EZH2 and Ki-67 expression and both in ameloblastoma and normal oral mucosa.Methods EZH2 and Ki-67 were detected by immunohistochemical SP method in 50 cases of ameloblastoma (30 cases primary,20 cases recurrence) and expression in 20 cases of normal oral mucosa.Results The positive expression of EZH2 protein in recurrent ameloblastoma was (36.25±7.24) ％,significantly higher than that of primary ameloblastomas (25.26±4.28) ％ (P ＜ 0.001).Ki-67 in recurrent positive expression in ameloblastoma was (34.96±5.28) ％,significantly higher than that of primary ameloblastomas (29.68±3.27) ％ (P ＜ 0.05).There was positively corTelated on EZH2 and Ki-67 (P ＜ 0.05),but not on the size of tumor and gender (P ＞ 0.05).Conclusions There are high expression of EZH2 and Ki-67 in ameloblastoma,and their play key role in the occurrence and development of ameloblastoma.They can be used as a reference index for the prognosis.The combined detection of EZH2 and Ki-67 protein expression with clinical evidence and its prognosis may contribute to the process of ameloblastoma and recurrence.

Objective To investigate the expression of NF-κBp65 in hippocampus after the XST intervention therapy in the SD rats with global cerebral I/R injury and testify the protective effect of XST after global cerebral I/R injury.Methods 72 healthy SD rats were randomly divided into 3 groups,sham operation(SO) group ( n =24),I/R group( n =24) and XST group( n =24).The model of acute global cerebral ischemia/reperfusion (including:I/R and XST group) injury was produced by means of simple Pulsinelli- brierley's four arteries occlusion method.H.E.staining was performed to detect the number of surviving neurons and TUNEL was used to detect the rate of neurons apoptosis.The expression activation of NF-κB p65 in hippocampus comu-ammonis ( CA1 ) region were examined by immunohistochemical method (SABC).Results The survival pyramidal neurons in the XST group continued to increase,and it was significantly more than the I/R group at each time-point after reperfusion[ (99.23 ±4.22)/mm vs (75.83 ±7.17 )/mm,(80.93 ± 5.36)/mm vs (51.50 ± 8.26 )/mm,(103.24 ± 5.48 )/mm vs (35.67 ± 13.17 )/mm,( 126.22 ± 7.54 )/mm vs (9.83 ± 4.71 )/mm ],the differences were statistically significant ( P ＜0.01 ).The apoptosis rate of pyramidal cell in the XST group at each time-point were more significantly reduced than the I/R group [ ( 8.82 ± 2.71 ) ％ vs ( 22.58 ± 4.68 ) ％.( 19.15 ± 6.23 ) ％ vs (42.68 ± 3.04 ) ％,( 11.82 ± 2.87 ) ％ vs ( 55.51 ± 6.81 ) ％,( 8.44 ± 3.23 ) ％ vs ( 71.69 ± 7.71 ) ％ ],the differences were statistically significant ( P ＜0.01 ).The positive neurons of NF-κBp65 expression in the XST group at different time-points were significantly less than the L/R group[ ( 13.20 ±2.50) vs ( 18.00 ± 1.87),(8.20 ±5.31) vs (41.60±3.65),(6.70±3.36) vs (55.30±5.10),(7.10±3.57) vs (72.80 ±4.71)],the differences were statistically significant ( P ＜ 0.05,P ＜ 0.01 ).Conclusions After global cerebral ischemia/reperfusion,XST could protect the brain from global cerebral ischemia/reperfusion injury by holding up the expression of NF- kappaB p65,and inhibiting neuronal apoptosis,and increasing the number of surviving neurons.Thus,the results of this experiment could provide a powerful and weighty objective indication for XST being used during cerebral resuscitation.

BACKGROUND:Previous studies have found that combined use of Buyang Huanwu decoction and bone marrow mesenchymal stem cel (BMSC) transplantation can play a synergic role against cerebral ischemia injury. OBJECTIVE:To analyze the effect of Buyang Huanwu decoction combined with BMSC transplantation to promote angiogenesis after cerebral ischemia. METHODS:Ninety-six Sprague-Dawley rats were randomly divided into four groups, and used to make middle cerebral artery occlusion models. In combined group, rats were given intragastrical administration of Buyang Huanwu decoction 10 mL/kg once a day, beginning at 3 days prior to modeling, and then given intragastrical administration of Buyang Huanwu decoction once at 2 hours after modeling, fol owed by its intragastrical administration every 12 hours. In BMSC and combined groups, BMSC suspension was injected into the rat ventricle after 2-hour cerebral ischemia/2-hour reperfusion, and then 30 minutes later, CD34 and CD45 antibodies were injected. In antibody group, CD34 and CD45 antibodies were injected. In model group, only normal saline was given. SerumαVβ3 level detection, immunohistochemical observation, Q-PCR and western blot tests were performed in the combined group at 12, 24, 36, 48 hours after reperfusion, while these indices were detected in the other three groups at 36 hours after reperfusion. RESULTS AND CONCLUSION:(1) The level of serumαVβ3 was lower in the antibody group than the model group (P<0.05), higher in the BMSC and combined groups than the antibody group (P<0.05), and higher in the combined group than the BMSC group (P<0.05). (2) Immunohistochemical findings showed that compared with the antibody group, the number of CD34 positive cel s was higher in the model, BMSC and combined groups (P<0.05). (3) Results from the Q-PCR and western blot assay showed that compared with the model group, the pFAK protein expression level was lower in the antibody and BMSC group, but FAK gene expression level had no overt changes;while the protein levels of FAK (24 hours after reperfusion) and pFAK (12 hours after reperfusion) were significantly increased in the combined group than the antibody and BMSC groups (P<0.05, P<0.01). Moreover, this increase exhibited a gradual y rising trend with the extension of reperfusion time. To conclude, the combined use of BMSC transplantation and Buyang Huanwu decoction can reverse the effect of CD34+CD45 antibodies that lead to the decrease in the number of vascular endothelial cel s and levels of integrinαVβ3 and downstream signaling molecules, thereby to promote angiogenesis in the MCAO model.

Objective To analyze the distribution of tophus in ankle and tarsal joints by multislice spiral computed tomography (MSCT)and to explore the pathogenesis of gout.Methods 34 patients with a documented diagnosis of gout underwent CT examina-tion of bilateral ankle and tarsal joints,and 55 ankle and tarsal joints were observed tophus.The sites of tophus deposition were cate-gorized into groups of ligament,tendon and joint in order to record and analyze.Results The tophus were showed at the anterior talofibular ligament (n=38),the medial ligament (the site of talus and calcaneus)(n=30)and the interosseous talocalcaneal liga-ment (n=29)(P <0.05)in the group of ligament;at achilles tendon (n= 18),followed by tibialis posterior tendon (n= 7)(P <0.05)in the group of tendon;at articulations intercuneiformes (n=1 5)and inferior tibiofibular articulation (n= 10)(P <0.05 )in the group of joint.Conclusion In ankle and tarsal joints,tophus is commonly deposited at the anterior talofibular ligament,at the achilles tendon and the articulationes intercuneiformes.

Objective To investigate the protective effect and mechanism of Astragaloside Ⅳ (AS-Ⅳ) on hypoxia/re-oxygenation induced by RAW264.7 murine macrophages.Methods The hypoxia/re- oxygenation induced RAW264.7 murine macrophages served as a model of I/R injury. The cells were divided into the control group, the model group, and the AS-Ⅳ group. The cell morphological changes of each group were observed directly under inverted microscope. Inducible nitric oxide synthase (iNOS), cluster of differentiation 206 (CD206), and peroxisome proliferator activated receptor-γ (PPAR-γ) were separately examined by RT-PCR, Western blot analysis and immunofluorescence staining.Results Compared with the model group, the numbers of cell in AS-Ⅳ group significantly increased , and the cell physiological status were much better. Compared with the model group, the iNOS immunofluorescence semi quantitative (0.62 ± 0.02 vs. 1.32 ± 0.09), the expression of mRNA (1.51 ± 0.07 vs. 3.46 ± 0.39), and protein (2.30 ± 0.14 vs. 5.16 ± 0.49) significantly reduced in AS-Ⅳ group (P<0.01). The CD206 immunofluorescence semi quantitative (1.01 ± 0.03 vs.0.61 ± 0.01), the expression of mRNA (0.91 ± 0.03 vs.0.51 ± 0.01), and protein (0.61 ± 0.04 vs.0.19 ± 0.01) significantly reduced in AS-Ⅳ group (P<0.01). Compared with the model group, the PPAR-γ immunofluorescence semi quantitative (0.60 ± 0.14 vs. 0.34 ± 0.03), mRNA (2.00 ± 0.14 vs.1.04 ± 0.03), andprotein (0.67 ± 0.05 vs.0.19 ± 0.01) significantly increased (P<0.01). Conclusions The AS-Ⅳ could attenuate ischemia-reperfusion injury by altering macrophages phenotype through upregulation PPAR-γ.

BACKGROUND:Mechanical strain certainly has an effect on physiological activities of osteoblasts. Runx-2 is a target of bone morphogenic protein signal and is an important factor for regulation of osteoblastic differentiation. Bone morphogenic protein signal transduction pathway is involved in physiological response of osteoblast to stimulation of mechanical centrifugal force.
OBJECTIVE:To observe the effect of mechanical centrifugal force on bone morphogenetic protein signal pathway under different time period and speed.
METHODS:MC3T1-E1 cells were pre-treated in DMEM medium containing 10%fetal bovine serum for 24 hours, and then divided into control group, 90 r/min group, 180 r/min group and 250 r/min group. Each group was then subdivided into 6 hours, 12 hours and 24 hours centrifugation subgroups. Experiments were repeated for three times for different centrifugal speed and different time period. Except centrifugation, the control group was under the same environment. Total RNA was extracted and reversely transcribed into cDNA. Runx-2 gene expression was determined by real time fluorescent quantitative PCR.
RESULTS AND CONCLUSION:The expression of Runx-2 mRNA was increased with extension of time, showing a positive correlation between the two. The mRNA expression at 180 r/min was significantly higher than that at 90 r/min and 250 r/min (P<0.01);at 90 r/min and 180 r/min, the Runx-2 mRNA expression was higher than that in the control group (P=0.039), both of them showed significant difference along with the time. The difference of centrifugal force speed and duration is associated with different physiological response of osteoblasts in bone morphogenic protein signal pathway, which plays an important role in mechanical signal transmission and cascade reaction.

Objective Studies show that the abnormal ex-pressions of APC and survivin play important roles in the development and progression of colon cancer .Survivin shRNA and APC double gene co-expression lentiviral vector was constructed to observe whether it could be successfully expressed in HT-29 colon cancer cells and whether it could impact on colon cancer cell apoptosis . Methods We selected the best shRNA interference fragment from the construc-tion of three pairs of complementary shRNA fragments and connected it with the effective fragment of APC ( aa1020-1698 ) to construct a double gene co-expression lentiviral vector .HT-29 cells were divid-ed into experimental group , empty loading group and blank group .HT-29 cells were observed by fluorescence microscopy after infec-tion.Survivin and APC expression levels were observed by real time PCR and western blot .Apoptosis was detected by caspase-3 activi-ty assay. Results ①We successfully constructed three pairs of shRNA sequences and proved that they had no human gene homolo -gous to the rest.Real time PCR analysis showed that the best sequence was shRNA 3.②After the sequence alignment of constructed shRNA vectors, three pairs of shRNA sequences were completely the same with the first designed sequence .Green fluorescence was observed in HT-29 cells by fluorescence microscope .The survivin content in experiment group (31.71 ±1.49) was significantly de-creased compared with empty loading group (100 ±0) and blank group(95.12 ±2.15)(P<0.05).The expression level of APC mR-NA was significantly increased compared with empty loading group (0 ±0) and blank group(0.51 ±0.15)(P<0.05).③The relative ratio of apoptosis in experiment group (0.573 ±0.050) was significantly increased compared with empty loading group (0.390 ± 0.040) and blank group(0.407 ±0.030)(P<0.05). Conclusion We have successfully constructed survivin-shRNA-APC double gene co-expression lentiviral vector which can be successfully expressed in HT-29 colon cancer cells , providing references for subse-quent gene therapy by the use of the carrier .