OBJECTIVE To investigate the effects and mechanism of PCB118 on cell-matrix and cel-cel adhesion in human hepatocel ular carcinoma cel s. METHODS Human hepatocel ular carcinoma cel s BEL-7402 were treated with PCB118 0.1,1.0 and 10.0 nmol · L-1 for 4 or 6 d,respectively. Then the cell-matrix adhesion assay and cell aggregation experiments were conducted to study the effect of PCB118 on cell-matrix adhesion and cell-cell adhesion in BEL-7402 cells. Quantitative real-time PCR and Western blotting methods were employed to assess the expression of key cytokines CD29,N-cadherin and E-cadherin. RESULTS The results showed that the cell-matrix adhesion ability of human hepato?cellular carcinoma cells BEL-7402 were significantly increased(P<0.05)after treatment with PCB118 0.1,1.0 and 10.0 nmol·L-1 for 6 d,whereas the cell-cell adhesion ability was significantly reduced(P<0.05). Exposure to PCB118(0.1,1.0 and 10.0 nmol·L-1)for 6 d induced significant upregulation of the mRNA expression levels of CD29 and N-cadherin along with the downregulation of E-cadherin(P<0.05). Western blotting analysis revealed that PCB118 exposure significantly increased protein expressions of CD29 and N-cadherin but reduced E-cadherin protein level(P<0.01). CONCLUSION PCB118 exposure affects the expression of CD29,N-cadherin and E-cadherin, which may be involved in PCB118-induced alteration of cell adhesion of hepatocellular carcinoma cell line BEL-7402.

The trypsin inhibitor is a kind of substance that can inhibit trypsin activity.It shares extensive physiological roles.The trypsin inhibitor not only inhibits the activities of many enzymes,but also has significant anti-cancer effects by suppressing cell invasion and promoting cell apoptosis.Wnt signaling pathway involves in the regulation of cell growth,proliferation and apoptosis.It also plays an important role in tumor development.This review focuses on the impacts of trypsin inhibitors on Wnt signaling pathway and tumor cell apoptosis.

Objective To investigate the anti-tumor effect and mechanism of curcumin in pancreatic cancer (PC). Methods Smad4 and Jab1 expressions were detected by immunohistochemistry in tumor tissues and pericarcinomatous tis?sue from 35 PC cases, and the correlation of Smad4 and Jab1 were analyzed based on the percentage of positive staining in?tissues from 21 random selected PC cases. The effect of curcumin on expressions of tumor suppressors p53, Smad4 and cell cycle inhibitor p27 were examined by Western Blotting after human pancreatic cancer cell line PANC-1 were divided into PANC-1 control group (no treatments were given) and PANC-1 curcumin group (treated with cell culture medium containing 10μmol/L curcumin). The effect of curcumin on expressions of combination of β-TrCP1 and Smad4 was examined by Co-Immunoprecipitation after human embryonic kidney cell line 293T were divided into 293T control group (no treatments were given), 293T curcumin group (treated with cell culture medium containing 10μmol/L curcumin) and 293T Jab1 group (trans?fected by HA-Jab1 plasmid). Results Compared with expressions in pericarcinomatous tissues, Smad4 was down regulated while the expression of Jab1 was upregulated in PC tissues (P<0.01), and the expression of Smad4 was negatively correlated with the expression of Jab1 (n=21, r=-0.71, P=0.007). After treated with curcumin, the protein expression of p53, Smad4 and p27 was increased in PANC1 cell, and the protein expression of the combination ofβ-TrCP1 and Smad4 was decreased in 293T cell (P<0.05). After transfected by HA-Jab1 plasmid, the protein expression of the combination ofβ-TrCP1 and Smad4 was increased in 293T cell (P<0.05). Conclusion Curcumin may have suppression effect of PC through increasing the protein expression of p53, Smad4 and p27, and the mechanism of Smad4 upregulation may be related with the inhibition of Smad4 ubiquitination process, while Jab1 may be also involved in Smad4 degradation through ubiquitination.

OBJECTIVE To explore the effect of clofenotane (DDT) on epithelial-mesenchymal transition (EMT) and the relevant molecular mechanism in human colorectal cancer cells. METHODS Human colorectal cancer cells DLD1 were treated with DDT 0.01, 0.1, 1.0, 10.0 and 100.0 nmol·L-1 for 48 h. Then, the morphology of DLD1 cells was observed. mRNA levels of E-cadherin, N-cadherin, vimentin and Snail1 were detected by real-time PCR. Protein expression of STAT3 signaling pathway of proteins STAT3 and p-STAT3 was detected by Western blotting. STAT3 inhibitor WP1006 (5μmol · L-1) was added to determine its impact on DDT-induced alternation of STAT3/Snail1 signaling and EMT-related molecules. Protein expression of STAT3 and p-STAT3 was detected by Western blotting and mRNA levels of E-cadherin, N-cadherin, Vimentin and Snail1 were detected by real-time PCR. RESULTS DLD1 cell morphology was changed after exposure to DDT 0.01-100.0 nmol · L- 1. Meanwhile, real-time PCR showed that the mRNA level of E-cadherin was significantly decreased compared with normal cell control (P<0.01), which was 42.4±2.8%of that in the normal control group. The mRNA levels of N-cadherin, Vimentin and Snail1 were significantly increased (P<0.01), which were 1.91±0.1, 1.5±0.2 and 1.5±0.1 times that of the normal control group. DDT 0.1, 1.0 and 10.0 nmol · L-1 exposure induced up-regulation of STAT3 and p-STAT3 protein levels (P<0.01), which were 2.1 and 1.8 times that of the normal control group. The addition of STAT3 inhibitor WP1066 (5 μmol · L-1) prevented STAT3 from phosphorylation as well as the up-regulation of Snail1(P<0.01), which was (56.3 ± 0.9)% that of the DDT 1.0 nmol · L-1 treat?ment group. Compared with DDT treatment alone, the mRNA levels of EMT-related molecules were remarkably reversed by WP1066 (5 μmol · L- 1) co-treatment, increasing E-cadherin but decreasing N-cadherin and vimentin in DLD1 cells(P<0.01), which were 50.2±2.9%and 61.6±6.1%of those in the DDT 1.0 nmol · L- 1 treatment group, respectively. CONCLUSION DDT alters the expressions of EMT-related molecules including E-cadherin, N-cadherin and vimentin via STAT3/Snail1 signaling, thus promoting the EMT process in human colorectal cancer cells. This progress may be closely related to DDT-induced colorectal cancer development.

Objective To investigate the effects of cyclooxygenase-2 inhibitor celecoxib on proliferation , invasion and migration of human pancreatic cancer cell line PANC-1 and then determine the optimal concentration of cele-coxib and the most suitable application time .Methods Human pancreatic cancer cell line PANC-1 was treated with diverse concentrations of celecoxib (20,60,100 μmol/L) for different durations (24,48,72 h).Cell prolifer-ation, invasion and migration capabilities were measured by MTT colorimetry , Transwell invasion assay , and scratch assay respectively .Results The proliferation capability of PANC-1 cell was reduced by celecoxib in a con-centration-and time-dependent manner ( P <0.05 ) .In addition , the invasion and migration capabilities were decreased by celecoxib in a concentration-dependent manner(P<0.01).Conclusions Celecoxib attenuates the proliferation of human pancreatic cancer cell line PANC-1 in a concentration-and time-dependent manner .Cele-coxib attenuates the invasion and migration in a concentration-dependent manner .

Objective To explore the expression of matrix metalloproteinase-14(MMP-14)protein in the human stomach carcinoma tissues and its correlation with carcinoma invasiveness and metastasis.Methods The MMP-14 protein expression was detected by immunohistochemistry in 59 cases of stomach carcinoma tissues (observation group)and 20 cases of normal stomach tissues (control group,the adjacent normal tissues from the tumor margin of 5 cm confirmed by pathology),and its correlation with the clinically pathological parameters was analyzed.The expression characteristics of MMP-14 among various TNM stages of stom-ach carcinoma were also analyzed.Results The positive rate of MMP-14 expression was 50.85%(30/59)in the observation group and 5.00% (1/20)in the control group,the positive rate of the observation group was significantly higher than that of the control group (P <0.01);the expression level of MMP-14 was correlated with the differentiation degree,regional lymph node metastasis degree,invasion depth,lymphatic invasion and TNM stage,which showing the statistical difference(P < 0.01);the expression of MMP-14 protein was up-regulated and showed the transferring trend from cytoplasm to cellular membrane along with the progres-sion of TNM stage.Conclusion The overexpression of MMP-14 protein exists in stomach carcinoma tissues,which contributing to the invasion and metastasis of stomach carcinoma cells.

Objective To study the effect of fat emulsion intravenous infusion on serum free fatty acids(FFAs) in rats.Methods 24 male Wistar rats were divided randomly into 3 groups,8 rats in each group.(1)Control group(NS),the rats were infused with normal saline intravenously and regular chow;(2)Group LCT,infused with 10% intralipid fat emulsion intravenously;(3)Group MCT/LCT,infused with 10% lipofundin fat emulsion. Group LCT and group MCT/LCT were continuously received equal calorie,nitrogen and volemin in 'All-in-One'solutions. Serum samples were drawn on the 8th day after PN for fatty acid determination. Results The FFAs in Group LCT and group MCT/LCT were remarkably higher than that in control group, but no difference between Group LCT and group MCT/LCT. Conclusions Fat emulsion intravenous infusion can increase the serum free fatty acids considerately.

OBJECTIVE To investigate the effect and mechanism of low concentrations of p,p′-dichlorodiphenyltrichloroethane (p,p′-DDT) on colorectal adenocarcinoma SW620 cell proliferation and apoptosis. METHODS SW620 cells were exposed to low concentrations of p, p′-DDT ranging from 1×10-12 to 1×10-6 mol.L-1 for 48 and 96 h. MTT assay was employed to examine the effect of p,p′-DDT on SW620 cell viability. Different cell stages of cycle and apoptosis rate were determined by flow cytometry after SW620 cells were exposed to 1×10-10 , 1×10-9 and 1×10-8 mol.L-1 for 96 h. Western blotting was used to determine the protein expression of Wnt/ β-catenin signaling components 〔β-catenin, phospho-β-catenin and phospho-glycogen synthase kinase ( GSK) 3β〕, their downstream target proteins ( c-Myc and cyclin D1)and apoptosis related proteins (Bcl-2, Bax, procaspase 8 and procaspase 3). RESULTS The viability of colorectal adenocarcinoma SW620 cells was significantly increased after exposure to low concentrations of p,p′-DDT ranging from 1×10-12 to 1×10-7 mol.L-1 for 96 h. p,p′-DDT 1×10-10 , 1×10-9 and 1×10-8 mol.L-1 exposure led to a decreased percentage of SW620 cells in G1 stage(P<0.01) along with an increased percentage of cells in S stage(P<0.01). Meanwhile, the apoptosis rate was signifi-cantly decreased compared with control group ( P < 0. 01). Exposure to p, p′-DDT from 1 × 10-10 to 1×10-8 mol.L-1 induced upregulation of phospho-GSK3β ( Ser9), β-catenin, c-Myc and cyclin D1 in SW620 cells( P <0.01). Moreover, apoptosis related proteins Bcl-2, procaspase 8 and procaspase 3 were unregulated(P<0.01), and Bax level and caspase 3 activity were decreased in p,p′-DDT-stimulated cells(P < 0.01). CONCLUSION Low concentrations of p, p′-DDT exposure activates Wnt/ β-catenin signaling and affects apoptosis-related proteins, which may be involved in p,p′-DDT-induced cell prolifer-ation as well as suppression of cell apoptosis in SW620 cells.

Objective To discuss the value of serum thymidine kinase 1 and DNA ploidy for the diagnosis of patients with acute myeloid lecukemia.Methods Determined TK1 and DI in 20 healthy people,6 patients with benign proliferate in hematological system and 66 patients with acute myeloid lecukemia by chemiluminescence detection technique and flow cytometry.Nonparametric comparisons among three group were done by rank sum test.ROC curve was used to determine the AUC and the diagnosis value serum thymidine kinase 1 and DNA.Results As showed by peripheral blood results,the TK1 (x2 =36.877,P＜0.001) and DI (x2=4.040,P＜0.05) had statistically difference among healthy people group,patients with benign proliferate in hematological system group and AML group.The normal control group compared with the AML group,TK1 (Z=-6.073,P＜0.001)and DI (Z=-2.012,P =0.044) had statistically difference;The normal control group compared with the benign proliferate patients,TK1 (Z=-1.234,P =0.169) and DI (Z=-1.084,P =0.278) had no statistically difference.The benign proliferate patients and that with AML patients,TK1 (Z=2.177,P=0.036) had statistically difference,but DI (Z=-1.801,P=0.061) had no statistically difference.The TK1 and DI area under the ROC curve were 0.950 (P＜0.001) and 0.638 (P=0.063),best cut-off were 1.73 pmol/L and 0.98,sensitivity were 0.95,0.78,and specifity were 0.88,0.39.Conclusion Serum TK1 and DI is a important diagnostic marker of early for AML patients,TK1 have a better diagnostic performance than DI significantly.

Objective To explore the characteristics of trunk muscle activity in persons with Parkinson's disease (PD) in search of better treatments for abnormal posture.Methods Ninety persons with PD of different severity and 8 normal controls were studied using surface electromyography (sEMG).A sixteen-lead sEMG instrument was used to collect sEMG amplitudes from the bilateral thoracic erector spinae,lumbar erector spinae,external oblique and rectus abdominis muscles when the subjects were lying,standing and walking.Results Compared with the control group,a significant increase was observed in the sEMG amplitude of the lumbar erector spinal muscle [(18.56±14.57) μV] and rectus abdominis muscles [(24.00±18.80)μV] of the PD group when lying.Significant relative increases in (P＜0.05),of the external oblique muscles were observed [(27.87± 11.49)μV] when standing (P＜0.05),and in the thoracic erector spinae muscle [(58.74±29.69)μV] and external oblique muscles [(59.01± 25.20) μV] increased when walking (P＜0.05).Compared with the control group,the sEMG activity of the external oblique muscles was significantly elevated in PD patients at Hoehn and Yahr stage 1 to 1.5 when walking.One or more of the trunk muscle groups showed significantly greater activity in the PD patients in Hoehn and Yahr stage 2 or 3 in all three positions.Conclusions The sEMG activities of the extensor and flexor muscles increase simultaneously in PD patients.More trunk muscles are involved in PD patients with higher H-Y grades.These findings provide a neurophysiological basis for the customizaton of rehabilitation therapy for patients with Parkinson's disease and for the precise selection of muscles for botulinum toxin injection.