Objective To evaluate the therapeutic effect of 89 Sr internal radiotherapy on multiple-bone metastasis of cancer. Methods Forty-nine cancer patients with multiple-bone metastasis received 89 Sr internal radiotherapy. The pain control effect, life quality improvement, change in levels of blood calcium and serum alkaline phosphatase (ALP), and side effects were analyzed respectively. Results The total effective rate of pain control was 77.6 %. The life quality was improved obviously. The levels of blood calcium and ALP were decreased. No obvious side effects were found during the treatment. Conclusion 89 Sr internal radiotherapy had a good therapeutic effect on multiple-bone metastasis of cancer.

Objective To construct a luciferase gene expression vector containing full-length 3' untranslated region(3'UTR)of mouse vascular endothelial growth factor-C(VEGF-C)gene,and to observe the effects of VEGF-C 3'UTR on luciferase gene expression by a double-fluorescence report system.Methods Polymerase chain reaction(PCR)was used to amplify VEGF-C 3'UTR and a 312bp VEGF-C coding region(CR)fragment from full-length VEGF-C cDNA in mouse Lewis lung cancer cells.The luciferase expression vectors containing VEGF-C 3'UTR or VEGF-C CR were constructed by subcloning the PCR products to luciferase reporter vector pGL3-Promoter using gene engineering technology,and then they were transfected to mouse Lewis lung carcinoma cells by LipofectamineTM 2000.The activities and mRNA expression of luciferase were detected by Dual-Luciferase Reporter System and quantitative RT-PCR,respectively.Results Mouse VEGF-C 3'UTR(429bp)and VEGF-C CR(312bp)were successfully amplified by PCR.The VEGF-C 3'UTR and VEGF-C CR fragments were successfully inserted into pTA2 vector,and then subcloned to pGL3-Promoter vector at Xba Ⅰ site by using restriction endonucleases analysis.The DNA sequences and insertion orientation of PCR products were all correct by sequencing analysis.The resulted luciferase expression plasmids were named pGL3-VEGF-C 3'UTR and pGL3-VEGF-C CR,respectively.Dual-Luciferase Reporter System detection and quantitative RT-PCR showed that in Lewis lung carcinoma cells,the activities of luciferase and expression of luciferase mRNA in the pGL3-Promoter group were higher than that in the pGL3-VEGF-C 3'UTR group,and there was no significant difference between pGL3-VEGF-C CR group and pGL3-Promoter group.Conclusion VEGF-C 3'UTR can inhibit luciferase gene expression.

Objective To explore whether alternatively activated macrophages (aaMphi) can transdifferentiate into lymphatic endothelial cells (LEC) under the inducement of VEGF-C, and to investigate the possible mechanisms involved in aaMphi-induced lymphangiogenesis. Methods An aaMphi model constructed by treating mouse macrophage cells RAW264.7 with mouse recombinant IL-4 for 24h was treated with different concentrations of recombinant mouse vascular endothelial growth factor-C (VEGF-C). After the clustering formed and the tube-like structures were detected in the matrigel, the aaMphi transdifferentiation system was finally decided to be constructed with the VEGF-C in the concentration of 100ng/ml. The mRNA expression of LEC specific markers, VEGFR-3 and Prox1, and aaMphi specific marker, Fizz1, were detected by real time quantitative RT-PCR on the 0, 7th, 14th, and 28th day, respectively, after VEGF-C stimulation. Formation of tube-like structure in the matrigel was observed with inverted phase contrast microscope in 28 consecutive days. Results The VEGF-C induced transdifferention system, incubated in EBM-2 medium and sustained by the matrigel, was successfully established. In this system, it was found that the mRNA expression of VEGFR-3 and Prox1 gradually increased, whilst that of Fizz1 decreased. The mRNA expression of VEGFR-3 and Prox1 reached the peak value, whilst that of the Fizz1 went down to the nadir, on the 14th day. No significant difference in values was found between the 14th day and the 28th day. During the period of the 7th day to the 28th day, distinct tube-like structures were gradually formed in the matrigel and the numbers increased in a time-dependant manner. Conclusion VEGF-C can induce the transdifferentiation of aaMphi into LEC by up-regulating the mRNA expression of VEGFR-3 and Prox1 in aaMphi, which is one of the possible mechanisms involved in aaMphi-induced lymphangiogenesis.

Objective To investigate the effects of active vitamin D (VD) on the expression of triggering receptor expressed on myeloid cells-1 (TREM-1) in renal tissue of diabetic nephropathies (DN) rats and to explore the impact of TREM-1 on adhesion and migration capacity of macrophage.Methods DN rat models were established by streptozotocin.Rats were randomly distributed into four groups:control (NC) group,VD group,DN group and DN+VD group (DN rats with 0.1 μg · kg-1 · d-1 calcitriol by garages).Rats were sacrificed respectively at 8 weeks and 12 weeks after treatment.Pathological changes in kidney tissue were detected and the expressions of CD68 and TREM-1 were acquired by immunohistochemistry stain and Western blotting.In vitro,RAW264.7 cells were divided into NC group,VD group,high glucose (HG) group and HG+VD group.In HG+VD group rats were treated by high glucose with 10-8 mol/L 1,25(OH)2D3.TREM-1 expression was measured by immunohistochemistry stain and Western blotting,and the ability of macrophage in migration and adhesion was evaluated by Transwell migration assay and adhesion assay.TREM-1 siRNA was transferred to silence TREM-1 expression,while plasmid of TREM-1 was transferred for high expression.Their ability of adhesion and migration in macrophage and the effect of 1,25(OH)2D3 were examined.Results (1) Compared with the NC group,the expressions of CD68 and TREM-1 were increased in DN group (P ＜ 0.05),whereas markedly decreased in DN+VD group (P ＜ 0.05).(2) The number of adhesion and migration cells,and the expression of TREM-1 protein in macrophage were obviously increased in HG group as compared with those in NC group (all P ＜ 0.05);whereas above changes were markedly decreased in HG+VD group than those in HG group (P ＜ 0.05).(3) The number of adhesion and migrated macrophage was reduced after TREM-1 siRNA intervention (all P ＜ 0.05).VD could significantly decrease the effect of high glucose on adhesion and migrated macrophages after TREM-1 siRNA (all P ＜ 0.05).(4) Adhesion and migration of macrophage were increased via TREM-1 overexpression (all P ＜ 0.05),but the effects of VD on high glucose-induced adhesion and migration of macrophage were disappeared.Conclusions VD can suppress the adhesion and migration of macrophage via reducing the expression of TREM-1,and inhibit infiltration of macrophage in renal tissue of DN rats.

Objective To investigate the effect of tumor necrosis factor ot (TNF-α) on monocyte-renal tubular epithelial cell adhesion and to explore its associated mechanism. Methods Human renal proximal tubular epithelial cell line (HK2 cells)and monocyte cell line (U937)were used to perform cell co-culture. TNF-α (10 μg/L)was used to stimulate HK2 cells and then U937 was put into the HK2 monolayers. Three locational parts of hyaluronan around HK2 cells were extracted by different ways and the HA expression was detected by enzyme-linked binding protein assay. Monocyte adhesion was detected by florescence recording assay. Flow cytometry was applied to assess intercellular adhesion molecule-1 (ICAM-1) expression in HK2 cells. Results Stimulation of HK2 cells with TNF-α significantly increased U937 binding to HK2 cell monolayer [(2.25±0.05) folds vs control, P<0.01]. Incubation of TNF-α (10 μg/L) with HK2 cells for 24 hours did not induce the change of total HA expression surrounding the HK2 cells, whereas HA redistribution happened with conditioned medium fraction (CM-HA) increased [(1.17±0.16) fold vs control, P<0.05], the pericellular fraction (trypsin extract fraction, TE-HA) decreased (83%±11% vs control, P<0.05) and the remaining cells-associated HA (cell associated fraction, CA) did not change significantly. In addition, pre-treatment of HK2 cells with TNF-α significantly increased the ICAM-1 expression [(1.85±0.22) folds vs control, P<0.01] and ICAM-1-dependent monocytes binding. Removal of HA from the surface of confluent monolayers of HK2 cells by hyaluronidase before addition of U937 cells increased monocyte binding[ (1.35±0.06) folds vs control, P<0.05]. In contrast, the presence of either sICAM-1 (400 μg/L) or anti-CDI8 antibody (10 mg/L) led to a significant decrease in ICAM-1-dependent monocyte binding (78%±7%, P<0.05; 75%±8%, P<0.01 vs those before adding sICAM-1 or anti-CD18 antibody, respectively). Conclusion HA redistribution induced by TNF-α may facilitate ICAM-1-dependent monocyte-epithelium binding, and pericellular HA plays a protective role in monocyte-driven tissue inflammation.

Objective To investigate the influence of Cordyceps polysaceharide (Cp) on epithelial-to-mesenchymal transition (EMT) induced by transforming growth factor-β1 (TGF-β1)in proximal tubular epithelial cells (PTEC). Methods HK-2 cell proliferation was determined by MTT assay. After incubation of HK2 cells with increasing concentrations of TGF-β1 (0, 0.1, 0.5, 1, 5, 10 μg/L) at 48 h and with TGF-β1 (5 μ/L) at different time points, E-cadherin, α-SMA, FN expression at transcriptional and protein levels were detected by real-time PCR and Western blotting respectively. The cells were pretreated with 1,5, 10 g/L Cp respectively for 24 h before adding TGF-β1 (5μg/L), then the cells were incubated for additional 48 h, mRNA and proteinexpression of above 3 cytokines was examined by real-time PCR and Western blotting as well. Results CP alone (0.01, 0.1, 1,5, 10 g/L) induced HK-2 cell proliferation in a dose-dependent manner. TGF-β1 enhanced α-SMA, FN expression while inhibited E-cedherin expression at both transcriptional and" protein level in HK-2 cell. At transcriptional level, compared to single TGF-β1 (5 μg/L) stimulating group, after Cp (1,5, 10 g/L) pretreatment for 24 h, the inhibition rate of a-SMA mRNA was 37.98%, 68.08% and 84.36%, respectively; FN mRNA was 46.97%, 63.82% and 81.85%, respectively; E-cadherin was up-regulated by 0.67 fold, 2.69 folds and 5.43 folds, respectively (P＜0.05). At protein level, the inhibition rate of α-SMA was 33.40%, 47.75% and 68.50%, respectively; FN was 16.26%, 65.92% and 80.30%, respectively; E-cadherin was up-regulated by 1.33 folds, 3.19 folds and 4.29 folds, respectively (all P＜0.05). Under Light microscopy, the Cp reversed cell shape from spindle-shape induced by TGF-β1 to nearly normal shape. Conclusion Cp may exert its inhibitive effects on TGF-β1-induced EMT.

Objective To investigate the influence of albumin on the expression of angiotensin-eonverting enzyme (ACE) in cultured human proximal tubular ceils (HK-2). Methods HK-2 cells were exposed to 2.5, 5, 10 g/L bovine serum albumin (BSA) for 6 h and 12 h respectively. The expression of ACE in HK-2 cells was detected by real-time RT-PCR and Western blot. Results Compared to the control group, the expression of HK-2 cells ACE mRNA treated for 12 h with different concentrations of BSA (2.5, 5, 10 g/L) significantly increased (P

Objective To explore the role of chemokine CXCL12 and its receptor CXCR4 in the directional migration of human prostate cancer(PCa).Methods The expression of CXCL12/CXCR4 in 18 human PCa samples and human PCa cell lines(PC3,DU145 and LNCap)was determined by immunohistochemistry and immunocytochemistry,respectively.Then the effect of CXCL12 on the migration and invasion of human PCa cell lines Was investigated by Matrigel invasion assay.Results Except 1 PCa sample,positive CXCR4 protein expression was detected in 17 clinical PCa samples.On the contrary,in 18 samples determined,only one sample expressed weak CXCL12 protein.CXCR4 rather than CXCL12 protein was exressed in PCa cell lines PC3,DU145 and LNCap.In addition,CXCL12 promoted the migration and invasion of PCa cell lines in a dose dependent manner in viiro,in which experiments PC3,LNCap cells were pretreated by antibody of CXCL12 or CXCR4 and then it was found the migrations of cells stimulated by CXCL12 were inhibited.Conclusion CXCR4 protein is expressed in human PCa and CXCL12/CXCR4 axis may play a significant role in the metastasis of prostate cancer.

Objective To investigate the effect of 1,25(OH)2D3 on high glucose induced podocyte injury and its signal transduction mechanism.Methods Differentiated mouse podocytes were exposed to normal glucose,high glucose,and different concentrations of 1,25(OH)2D3 or LY294002 (a selective PI3K inhibitor) for 24 h.PCR and immunofluorescent staining were used to detect nephrin,podocin,and desmin.Western blotting was used to detect protein expression of nephrin,podocin,desmin,PI3K,Akt and p-Akt.Results Compared with high glucose group,1,25(OH)2D3 (100 nmol/L and 1000 nmol/L) significantly up-regulated the expression of podocin and nephrin in podocytes induced by high glucose (P ＜ 0.05).Meanwhile,1,25(OH)2D3 (100 nmol/L) significantly reduced the expression of desmin (P ＜ 0.05).PI3K and p-Akt were obviously reduced in high glucose group.In the presence of 1,25(OH)2D3,the trends were reversed.However the above effects of 1,25(OH)2D3 were abolished when p-Akt was blocked by the PI3K inhibitor LY294002.Conclusions 1,25 (OH)2D3 can inhibit high glucose-induced pedocyte injury through PI3K/p-Akt signaling pathway.

Objective To investigate the risk factors of chylous leakage after pancreatioduodenectomy so as to find effective measures to prevent this complication.Methods A retrospective analysis was conducted on 230 patients who underwent pancreatioduodenectomy at the First Affiliated Hospital of Zhejiang University from Jun.2012 to Jun.2014.Patients with chylous leakage were identified and a 1 ∶ 2 patients in the study and the control groups were selected.The parameters for matching included tumor volume,vascular invasion,and extent of lymph node dissection.A logistic analysis was performed to identify independent risk factors of chylous leakage.Results 15 (6.5％) patients developed chylous leakage after pancreatioduodenectomy.The average hospital stay after surgery of the study group was 20.8 days,compared to 13.5 days in the control-group (P =0.004).In the study group,chylous leakage rate increased in patients with 14th and 16th group of lymph nodes dissection (80％ vs 36.7％,P =0.006).Logistic analysis showed that 14th and 16th lymph nodes dissection was an independent risk factor of chylous leakage after pancreatioduodenectomy (P ＜ 0.05,OR =6.909,95％ CI 1.593 ～ 29.958).Conclusions Chylous leakage prolonged hospitalization after pancreatioduodenectomy.Dissection of the 14th and 16th lymph node groups was an independent risk factor of chylous leakage after pancreatioduodenectomy.Careful ligation of the gastrocolic vein near the lymphatic trunk and dissection of 14th and 16th group of lymph nodes were effective interventions to reduce postoperative chylous leakage.