[ ABSTRACT] AIM:To study the influence of Raptor on the invasion ability of glioma cells.METHODS: The technique of RNA interference was used.U87 cells were transfected with Raptor restricted siRNA plasmid, and the expres-sion level of Raptor in the transfected cells was detected by Western blotting.The invasive ability of the cancer cells in vitro was determined.The phosphorylation level of ARK5 and the expression of MMP-2 and MMP-9 were detected by Western blotting.The expression levels of Raptor in the tumor samples of low-grade gliomas ( WTO grade I and grade II) and high-grade gliomas (WTO grade III and grade IV) were also analyzed by immunohistochemical staining.RESULTS: Raptor siRNA was transfected into U87 cells and the cells were named siRaptor/U87 cells.The cells transfected with the control plasmid was named Scr/U87 cells.The expression level of Raptor in siRaptor/U87 cells was lower than that in Scr/U87 cells.The results of in vitro invasion assay showed that the number of siRaptor/U87 cells penetrating the Matrivgel matrix membrane was less than that of Scr/U87 cells (P<0.01).The protein expression of MMP-2 and MMP-9, and phosphoryl-ation of ARK5 protein in the cells in the experimental group were lower than those in control group.The correlation between the expression of Raptor in gliomas and the degree of deterioration was also observed ( P<0.01) .CONCLUSION: The expression of Raptor may contribute to the invasion ability of glioma cells by phosphorylation of ARK5 and increase in the levels of MMP-2 and MMP-9.

Objective To study the diagnostic value of radial echoendoscopy combined with ultrasonic micro-probe for differentiating mediastinal disease from esophageal submucosal lesion.Methods Data of 53 patients with esophageal eminence with unknown origin detected by gastroscopy or CT in China-Japan Union Hospital were reviewed.Ultrasonic micro-probe was performed first and followed with radial echoendoscopy.The diagnosis was compared with the results of CT.Results The diagnostic sensitivities of EUS and CT for esophageal submucosal lesion were 90.9％ (20/22) and 50.0％ (11/22),and the specificities were 93.5％ (29/31)and 83.9％ (26/31).The sensitivities for the benign mediastinal disease were 91.7％ (22/24) and 70.8％ (17/24),and the specificities for the benign disease were 93.1％ (27/29)and 62.1％ (18/29).The sensitivity and specificity for malignant mediastinal disease were both 100.0％.Conclusion Radial echoendoscopy combined with ultrasonic micro-probe shows prominent advantage in the diagnosis of the esophageal submucosal lesion and mediastinal disease over CT,especially in the specificity and sensitivity for esophageal submucosal lesion and benign mediastinal disease.

Purpose To investigate the correlation of Dock180 and miR-31 expression in breast cancer cells,and to observe the effect of miR-31 on the invasion of breast cancer cells by Dock180.Methods MiR-31 was transfected into breast cancer cells by liposome transfection technique.The actual binding site of miR-31 to the 3'-untranslated region of Dock180 was confirmed through luciferase assay.Western blot was performed to detect the expression of Dockl80 and other related proteins.Real-time PCR was used to measure the expression of Dock180.Matrigel invasion were performed to detect the invasion of breast cancer cell lines with miR-31 increased.Resuits The protein levels of Dock180 in breast cancer cell lines negatively correlated with miR-31 expression,and Dock180 was directly targeted by miR-31.Dock180 downregulation and miR-31 overexpression reduced breast cancer cells invasion.Conclusion Dock180 modulated by miR-31 plays an important function in breast cancer cell lines invasion.

0.05) . In the later stage (8～12 weeks), the tissue reaction nearly subsided in PLGA stented ureters after degradation of the device. Whereas, the tissue reaction induced by UROVISION stent had lasted throughout the observation period, even deteriorated with time going(P

Objective:This study aimed to investigate the effect and significance of a binding protein-2 (Gab2)-Akt-ARK5 signaling pathway on the invasion of glioma cells. Methods:Immunohistochemical methods were used to detect the expressions of Gab2 and ARK5 in 45 cases of glioma tissue. siRNA plasmid was used to transfect LN-229 cells, and western blot was performed to analyze the protein expressions of Gab2 and ARK5. In vitro Matrigel invasion assay was conducted to detect variations in the invasiveness of transfected cells. Western blot was also conducted to analyze the protein phosphorylation of Akt and ARK5 in the cells transfected with Gab2 plasmid. Results:Immunohistochemical assay revealed that the expressions of ARK5 and Gab2 in glioma cells were positively correlated, and both expressions were higher in high-grade glioma (WHO gradeⅢ,Ⅳ) than in low-grade glioma (WHO gradeⅠ,Ⅱ). LN-229 cells transfected with ARK5 plasmid, Gab2 plasmid, ARK5 and Gab2 plasmid, and control plasmid were named siARK5/LN-229, siGab2/LN-229, siARK5 and siGab2/LN-229, and SCR/LN-229, respectively. After transfection was performed, the protein expressions of ARK5 and Gab2 were respectively decreased in siARK5/LN-229 and siGab2/LN-229. The protein expressions of ARK5 and Gab2 in siARK5 and siGab2/LN-229 were also respectively decreased. After ARK5 or Gab2 was downregulated, the number of glioma cells, which invaded and penetrated Matrigel, was decreased (P<0.01). The number of glioma cells also decreased significantly after ARK5 and Gab2 were downregulated. The phosphorylation of Akt and ARK5 in siGab2/LN-229 cells was decreased after these cells were stimulated by insulin-like growth factor-1. Conclusion:The silencing of ARK5 or Gab2 impaired glioma cell invasiveness. The decreased protein expression of Gab2 inhibited the phosphorylation of Akt and ARK5. These results suggested that the Gab2-Akt-ARK5 signaling pathway could be relevantly involved in glioma cell invasion.

Background and purpose:Nir1 is a transmembrane receptor for chemokine (C-C motif) ligand 18. CCL18 speciifcally binds to Nir1 at the cellular membrane of breast cancer cells to exert its invasion and metastasis. However, the speciifc mechanism of Nir1 is not clear in glioma. This study probed the effect and mechanism of Nir1 in the invasion of glioma cells.Methods:Western blot was used to detect the expression of Nir1 in glioma cells. siRNA plasmid was used to transfect U251 cells. Western blot was used to analyze the expression of Nir1 and protein phosphorylation of Akt in the cells transfected by Nir1 plasmid.In vitro Matrigel invasion assay was used to detect the invasive ability in the cells that were transfected. F-actin polymerization assay was used to detect F-actin recognition ability in cells.Results:The expression of Nir1 was higher in all glioma cells. After transfection, the invasion of siNir1/U251 was obviously decreased than the SCR/U251, F-actin content was reduced compared to the control group. Akt phosphorylation experiment result showed that the protein phosphorylation of Akt was enhanced in control group cells CCL18 following stimulation. However, the existence of CCL18 would affect the phosphorylation of Akt in siNir1/U251.Conclusion:Nir1 is high expression in glioma cells, and Nir1 binding to chemokine CCL18 promotes glioma cells invasion and metastasis through regulation the phosphorylation of Akt and F-actin polymerization .

Purpose To investigate the expressions of PKCζ, MMP-2, and MMP-9 in breast cancer and the relationship with the inva-sion and metastasis of breast cancer. Methods The expression of PKCζ, MMP-2 and MMP-9 in 100 cases with breast cancer was as-sessed with immunohistochemistry PV 9000 method. PKCζ-siRNA was transfected into MDA-MB-231 cell lines, called siPKCζ/MDA-MB-231. While siRNA construct containing a scrambled sequence was transfected into MDA-MB-231 cells to generate control cells, which were designated as Scr/MDA231 cells. Western blotting was used to measure the expression of PKCζ in transfected cells, and the Transwell invasion assay was used to detect the invasion ability in vitro. The content of MMP-2, MMP-9 were measured by ELISA. Results The expression rates of PKCζ, MMP-2 and MMP-9 in breast cancer tissues were 62.5%, 37.5% and 32.5%, and there were significant differences among them (P<0.05). The expression of PKCζwas much higher than those in the normal breast tissues nearby. The expression of PKC protein was assoiated with lymph node metastasis, distant metastasis (P<0.01), but was not correla-ted with other clinicopathologic parameters, such as age, tumor size, histological type, ER, PR, and so on (P>0.05). The expres-sion of PKCζ, MMP-2 and MMP-9 were lower in siPKCζ/ MDA-MB-231 group than those in scr/ MDA-MB-231 group, and the in vitro invasion ability was significantly decreased (P<0.05). Conclusions PKCζ can promote the invasion and metastasis of breast canc-er, and correlated with the expression of MMP-2, MMP-9(P<0.05).

Objective To observe the effects of Qingjiefang on insulin resistance and reduction of microalbuminuria in early diabetic nephropathy.Methods A total of 64 patients with early diabetic nephropathy were randomly recruited into a control group and a treatment group.The control group(32 cases)was treated with conventional western medicine,while the treatment group(32 cases)was treated with Qingjiefang based on the control group.The changes of the fasting plasma glucose(FPG),2hBG,glycosylated hemoglobin(HbAlC),fasting Insulin(FINS),24 hours urinary albumin excretion rate (UAER).body mass index(BMI)and insulin resistance(IR)were observed before and after the treatment.Results After the treatment,FINS,IR and UAER were decreased in the treatment group,the comparison with the control group showed significant difference (P<0.05).Conclusion Qingjiefang can effectively improve insulin resistance and prevent early diabetic nephropathy.

Background and purpose:Numerous researches indicated that the expression of pituitary tumor transforming gene1 (PTTG1) was correlated with the severity of glioma tumors. However the specific mechanism of PTTG1 is not clear in glioma. In this study, we explored the role and significance of PTTG1 in the invasion of glioma cells. Methods:Western blot was used to detect the expression of PTTG1 protein in various glioma cell lines. siRNA plasmid was used to transfect U87 cells. Western blot was used to analyze the expression of PTTG1 protein in transfected U87 cells. Matrigel invasion assay was used to detect the invasive ability in the cells being transfected in vitro. Western blot was used to analyze epithelial growth factor (EGF) induced protein phosphorylation of ARK5 and Akt in the cells being transfected PTTG1 plasmid (siPTTG1/U87) and scrambled siRNA (Scr/U87). Results:The expression of PTTG1 protein was higher in all glioma cell lines. After transfection, the invasion of siPTTG1/U87 was obviously decreased after 5 min with EGF stimulation than the Scr/U87, the phosphorylation of ARK5 and Akt was significantly enhanced. However, whether or not the existence of EGF, the phosphorylation of ARK5 and Akt had no differences in siPTTG1/U87. Conclusion:In glioma cells, PTTG1 protein is high expressed and maybe have an important function in glioma cells invasion through Akt-ARK5 signaling pathway.

Facing the changes in employment of pathology postgraduates and needs of clinical applied pathology talents,this paper discussed on problems in clinical practice ability cultivation of pathology postgraduates as well as elaborated on cultivation mode for clinical applied pathology postgradu-ates from the aspects of teaching style, training of clinical practice ability and assessment methods and by taking exploration-and-discussion teaching as principal line.