Objective To investigate the expression of NF-κB and epithelial-mesenchymal transition (EMT) related markers E-cadherin and Vimentin proteins in human pancreatic cancer tissues and its relation with the malignant features. Methods The expression of NF-κB、E-cadherin and Vimentin proteins in 62 cases of pancreatic cancer tissues were detected by using immunohistochemistry and compared with the clinicopathological data of pancreatic cancer. Results The positive expression rate of NF-κB was 81% (50/62), Vimentin protein increased of expression was 61% (38/62), and E-cadherin protein loss of expression was 55 % (34/62) in pancreatic cancer. The positive expression rate of NF-κB was significantly related with the lymph node metastasis (x2=11.761, P＜0.05), distant metastasis (x2=9.225, P＜0.05), the absent expression of epithelial marker E-cadherin protein (r =0.352, P ＜0.05) and the positive expression of mesenchymal marker Vimentin protein (r=0.343, P ＜0.05), but there was no relation with the patients gender,age, tumor location, tumor type and tumor differentiation (P ＞0.05). In addition, the significant correlation of E-cadherin expression loss and Vimentin expression with tumor lymph node metastasis and distant metastasis was found (x 2= 6.914, 4.984, 7.753, 5.144, P ＜0.05). Conclusion The overexpression of NF-κB in pancreatic cancer may accelerate invasion and metastasis of pancreatic cancer through inducing EMT.

Objective To investigate the inhibitory effect of immunosuppressive agent FRY720 on hepatocellular carcinoma Hepal-6. Methods Hepal-6 cells were cultured, and divided into 4 groups, namely control group and 0.1 μg/ml, 10 μg/ml, 100 μg/ml quality concentration groups. The cells were treated by the drugs for 24 to 48 hours respectively. The inhibitory rate of the cells was measured by MTT assay, and cell cycle and cell apoptotic rate were detected by flow cytometry (FCM). Results The ability of tumor cell growth were inhibited by FTY720 after 48 h. The maximal inhibition rate was 62.10 %, The apoptosis ratio was increased when FTY720 was 0.1-100 μg/ml, and it was 4.07 %, 8.16 %, 19.84 % respectively. FTY720 significantly prolonged cell G1 phase. Conclusion FTY720 could inhibit the growth of hepatocellular carcinoma, arrest the cell in G1 phase, and increase apoptosis.

Objective To investigate the clinical application of Morse Fall Scale by primary nurses.Methods Morse Fall Scale was used to assess the falling risk in 21,378 patients by the primary nurses.The application of the Morse Fall Scale, choices of right time for assessment and the occurrence of falls in patients with risks of falling were evaluated by a self-designed questionnaire.Results Among 21,378 patients, 222 patients were at risk of falling.In terms of the items like the history of falls,more than one diagnosis,intravenous injection,the accuracy of assessment was above 91.6%.In terms of the items like aided walking,gait,cognitive state,the assessment accuracy is below 86.0%.The accuracy of assessment timing selection in the situations like admission,the score by Morse fall scale over 24at admission and the accuracy in choosing right time for assessment was was above 90.2% after transferring to other deparments and falling.The accuracy in choosing the right time for fall assessment was below 67.9% under such conditions as apostasies,and oral administration of drugs probably leading to fall. Conclusions It is acceptable that the primary nurses use the Morse fall scale to assess patients with risky falls and for the right choice of assessment.Attention should be paid to strengthen the nurses training to improve the accuracy of the scale,to make sure to choose right time for the assessment so as to prevent the falls of patients to the greatest extent.

Objective To study the action of CNOT 7 (CCR4-NOT transcription complex subunit 7 human)gene and its mechanisms in the process of Vγ 9Vδ2T cell immunologic tolerance of HepG2 cells (Hepatoblastoma G2 Cell Line).Methods The shCNOT 7 (Recombinant plasmid of CNOT 7) and control vector shRNA were transfected into HepG2 cells.Vγ9Vδ2T cytokine stimulated each group before and after cell transfection,Cell apoptosis was detected by flow cytometry (FCM),CNOT 7 protein and STAT1,STAT3 expression level was detected by Western blot.CNOT 7,STAT1 and STAT3 protein expression levels of HepG2 liver cancer cell lines and L02 normal liver cell line was assayed by Western blot.Results When stimulated by Vγ9Vδ2T cytokine,the apoptosis rate of gene-knockdown group significantly improved from (7.55％ ±2.63％) to (20.59％ ±3.12％).Compared with L02 cells,the CNOT7 protein expression of HepG2 cells increased (F =28.76,P ＜ 0.01),STAT3 protein expression increased (F =110.29,P ＜ 0.01),while STAT1 protein expression was down-regulated (F =35.67,P ＜ 0.01).CNOT 7 knockout could induce HepG2 cells STAT1 expression (t =6.69,P ＜ 0.05).Conclusions CNOT 7 gene could induce HepG2 cells Vγ 9Vδ2T cellular immune tolerance.CNOT 7 knockout could reverse the Vγ 9Vδ2T cell immunologic tolerance of HCC.

Objective To investigate the significance of CerbB-2 oncogene in the occurrence and development of gastric adenocarcinoma. Methods By using immunohistochemistry and fluorescence in situ hybridization, 60 cases of gastric adenocarcinoma tissue, 60 cases of normal margin of mucosal tissue and 30 cases of normal gastric mucosa tissue were detected for the expression and amplification of CerbB-2. Results The positive rate of CerbB-2 expression in gastric adenocarcinoma tissues, normal margin of mucosal tissues and normal gastric mucosa tissues were 31.7 %, 8.33 % and 3.33 %; The positive rate of CerbB-2 amplification of these were 28.3 %, 5 % and 0. There were significant differences between gastric adenocarcinoma tissues and the others in the expression and amplification of CerbB-2. The expression and amplification of CerbB-2 in gastric adenocarcinoma tissues had no correlation with age,gender,tumor size or tumor differentiation degree of patients' (P >0.05), but were correlated with the extent of tumor invasion, lymph node metastasis, distant metastasis and clinical stage(P <0.05). Conclusion CerbB-2 is closely related to the occurrence and development of gastric adenocarcinoma, and can be considered as an important index for determining the biological behavior of gastric cancer.

Objective To study the relationship between Treg cell numbers and level of TGF-β1,IL-10 in the immune microenvironment of rat liver cancer.Method 40 male Wistar rats were randomly divided into three groups,liver cancer model group (n =20),saline control group (n =10) and blank control group (n =10).Liver cancer model was established by intraperitoneal injection of DEN (100 mg/kg).Percentage of Treg cells in cancer tissues was detected by fiow cytometry and the expression level of TGF-β1 and IL-10 by immunohistochemical SABC.Results The ratio of Treg cells in CD4 + T cells was 14.32％ ± 4.84％ in liver cancer tissues,significantly higher than that in the blank control group 5.64％ ±6.10％ and the saline control group 7.95％ ±3.55％.The difference was statistically significant (F =211.279,P ＜ 0.05).The expression level of TGF-β1 and IL-10 in cancer tissues was significantly higher than the blank control group and the saline control group (FTGF-β1 =250.740,FIL-10 =152.744,all P ＜0.05).The number of Treg cells was positively correlated with TGF-β1,IL-10 level (P ＜ 0.05).Conclusions TGF-β1,IL-10 and Treg cells significantly increased in rat experimental liver cancer tissues,and there was positive correlation between Treg cells,and TGF-β1 and IL-10 in liver cancer tissues.

Objective To analyze the differentially expressed genes between the NCAM + c-Kit +RBE and NCAM-c-Kit-RBE of intrahepatic cholangiocarcinoma (ICC) cell lines,and to screen out the differentially expressed genes that are related to the stem cell signaling pathways.Methods Magnetic activated cell sorting was used to isolate the NCAM + c-Kit +/NCAM-c-Kit-subset cells,and then Agilent Whole Human Genome Microarray Kit was used to test the difference in gene expressions between the NCAM + cKit + and NCAM-c-Kit-subset cells.The difference in gene expressions related to the stem cell signaling pathways was analyzed by the SAS system.The result of the microarray was further confirmed by RT-PCR.Results The total differentially expressed genes which could be found through gene microarray were 7270 [foldchange(fc) ≥2 or fc ≤0.5].Compared with the NCAM-c-Kit-RBE,3572 genes were upregulated while 3698 genes were downregulated.The differences in gene expressions related to the stem cell signaling pathways were 421 (fc ≥2 or fc ≤ 0.5),among which 231 genes were upregulated while 190 genes were downregulated.Conclusions High-flux microarray could be used to screen out lots of differentially expressed genes between the NCAM + c-Kit + and NCAM-c-Kit-RBE cells.The differences in gene expression in the stem cell signaling pathways could also be further analyzed using the SAS system.

Objective To investigate the percentage,mature classification and Immune killing function of Vγ9Vδ2T cell in peripheral blood of HCC patients.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from HCC patients (n =25) and healthy donors (n =20) by discontinuous density gradient centrifugation.Proportion,mature and differentiate subtypes and IFN-γ and CD107a expressing of the delta 2 T cells were detect by using flow cytometry,δ2Tcell were selectively cultured with zoledronate and human IL-2.After 12-14 days cells were collected and tested for the second time.Results While the percentage of Vγ9Vδ2Tcell of total T cell in peripheral blood of HCC patients is lower than healthy people before culture (t =4.505,P ＜ 0.001),after augmentation in vitro the proportion increased significantly (t =8.782,P ＜ 0.001),to a level similar to healthy group (t =1.644,P =0.109).There was no statistically significant difference when differentiation subtypes of patient's Vγ9Vδ2Tcell were compared with healthy group before culturing (all P ＞ 0.05),after culture the proportion of Tn,Tcm and Temra decreased [t(Tn) =2.081,t(Tcm) =2.478,t(Temra) =2.953,all P ＜ 0.05],and the proportion of Tem,Tem+ Temra increased [t(Tem) =12.6,t (Tem + Temra) =9.843,all P ＜ 0.001].Cell culture did not alter the proportion of IFN-γ and CD107a secreting Vγ9Vδ2T cells in the peripheral blood in both HCC patients and healthy people (all P ＞ 0.05).Conclusions While the percentage of Vγ9Vδ2T cell of HCC patients in peripheral blood was lower than healthy people,its matured subtypes are similar to those of healthy people,and functions of expressing IFN-γ and CD107a are not different with healthy people.Applying ZOL + IL-2 can amplifyVγ9Vδ2T cells of patients with HCC.

AIM:To study the influence of stimulation by LPS and CD40 ligandization in vitro on the TLR4-MD2 expression and IL-12 production in dendritic cells (DCs) modified by sCD40 gene and provide the experimental clues of inducing dornor-specific immune tolerance.METHODS: Plasmid pEGFP-N1/sCD40 and pEGFP-N1 was transfected into DC2.4 cell line with lipofectamine. After 6 h of treatment with LPS and anti-CD40mAb, the expression of TLR4-MD2 on DCs was determined with flow cytometry (FCM) and RT-PCR. IL-12p70 protein was detected by ELISA.RESULTS: LPS treatment of DCs down-regulated surface expression of TLR4-MD2, LPS treatment along with anti-CD40mAb significantly up-regulated TLR4-MD2 surface expression. CD40 ligandization did not affect TLR4-MD2 mRNA expression in DCs but partly increased its low level induced by LPS and markly enhanced IL-12p70 secretion after LPS stimulation. DCs modified by sCD40 gene inhibited the above effect.CONCLUSION: After treatment with LPS and anti-CD40mAb, DCs modified by sCD40 gene down-regulate surface expression of TLR4-MD2 and IL-12p70 secretion decreases significantly, which might be linked with the interruption of TLR4-MD2 transportation from cytoplasm.

Objective To analyze the clinical effect of pedicle screw fixation in the treatment of multi-segmental lumbar fracture and dislocation under the guidance of visualization technique. Methods A total of 21 patients with multi-segmental lumbar fracture and dislocation were selected from November 2012 to November 2013. Before the screw implantation, the structure of bilateral pedicle was observed through Mimics software and the implantation parameters were measured. The position of pedicle screws by postoperative CT scan, operation time, and the satisfaction of the patients were assessed. The percentages of anterior vertebral height and Cobb′s angle were measured before operation, 2 weeks and 8 months after operation. Results All patients were satisfied with informed consent score and the way of pedicle screw and the selection of plant were more reasonable. With better screw position, shorter operative time and less blood loss and adverse reactions, pedicle screw fixation achieved good effect. Conclusion With high security and considerable clinical value, pedicle screw fixation in the treatment of multi-segmental lumbar fracture and dislocation under the guidance of visualization technique has exact and good effecct.