Circulating vascular endothelial growth factor-C (VEGF-C) and vascular endothelial growth factor (VEGF) levels in patients with colorectal carcinoma were determined in order to assess their clinical significance as a diagnostic tool for monitoring lymph node metastasis. In 66 patients with colorectal carcinoma and 30 healthy controls, circulating VEGF-C and VEGF levels were assessed by using enzyme-linked immunosorbent assay (ELISA). Serum VEGF-C and VEGF levels were higher in patients with colorectal carcinoma than in healthy controls. Patients with lymph node metastasis had higher serum VEGF-C and VEGF levels than those without lymph node metastasis. The levels of VEGF-C and VEGF were higher in the invasion group than in the non-invasion group. Serum VEGF-C levels reached a sensitivity of 81% and a specificity of 76% with a cutoff value of 1438.0 pg/mL, whereas VEGF levels reached 72% sensitivity and 74% specificity at 240.2 pg/ mL. If 66 patients were divided into 4 groups according to the combined determination of VEGF-C and VEGF levels, the positive predictive value was 85.3%, the negative predictive value was 94.6%, and accuracy was 93.7%. It was suggested that circulating VEGF-C levels might provide additional information for distinguishing the absence from presence of lymph node metastasis in patients with colorectal carcinoma. The combined determination of VEGF-C and VEGF levels could be used as an important index for preoperatively clinical stage of colorectal carcinoma.

Objective To study the relationship between the expression of chemokines monocyte chemoattractant protein 1 (MCP 1) and the biological behavior of colorectal carcinoma by detecting chemokine mRNA and protein in human colorectal cancerous tissues. Methods The expression of the MCP 1 mRNA was detected in colorectal carcinoma collected freshly from surgical specimens by RT PCR and the expression of the MCP 1 protein was assessed in colorectal carcinoma tissues collected from surgical specimens by immunohistochemistry. Results All the 12 colorectal carcinoma expressed the MCP 1 mRNA, which was detected by RT PCR; MCP 1 protein was detected in 90% of the tumors; The expression of the MCP 1 protein in colorectal carcinoma was correlated with its state of metastasis and the Dukes' stage. Conclusions The expression of chemokine MCP 1 in colorectal carcinoma may influence its biological behaviour.

Objective To evaluate the diagnostic value of preoperative MRI for advanced gastric cancer and metastasis. Methods MRI was preoperatively performed in 36 patients with advanced gastric carcinoma by the superconductive type of magnetic resonance imaging instrument through a single breath holding fast scan sequence, the results were correlated with the intraoperative and histopathologic findings.[WT5”HZ] Results[WT5”BZ] The correct rate of localization and qualitative determination in gastric carcinoma by MRI were both 100%. The displaying rate of metastatic lymph notes was 83%(25/30), and the correct diagnostic rate was 73%(35/48) for metastasis or invasion to adjacent abdominal organs.[WT5”HZ] Conclusions[WT5”BZ] Gastric carcinoma shows characteristic MRI pictures, hence MRI examination has the advantage of diagnosing gastric cacinoma, its spreading, even metastasis of lymph nodes.

Objective To screen for the differentially expressed genes in breast cancer cell line MCF 7 and tamoxifen resistant breast cancer cell line LCC2 by using cDNA microarray Methods The PCR products of 8000 genes were spotted on chemical material coated glass plates in array The DNAs were then fixed on the glass plate by a serial of treatments The total RNAs were isolated from cells cultured in the flash, and then were purified to mRNA by Oligotex Both the mRNAs from different breast cancer cell lines were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP, for preparing the hybridization probes The mixed probes were then hybridized to the cDNA microarray After high stringent washing, the cDNA, microarray was scanned for the fluorescent signals and showed the differences between the cell lines Results Among the 8?000 target genes, there were 1?892 (23 65%) genes expressing differently between MCF 7 and LCC2 cell lines Bioinformational analysis on those genes was performed Conclusion DNA microarray technology is an effective technique in screening for differentially expressed genes between different tissues and cell lines

The effects and mechanisms of exogenous phosphatase and tensin homolog deleted from chromosome ten (PTEN) gene on phosphatase activity-dependent apoptosis of breast cancer cell line MDA468 were investigated. PTEN gene packaged with lipofectin was transferred into breast cancer cell line MDA468 and parental MDA468 cells served as controls. RT-PCR and Western blot were done to detect the expression of target genes. The expression of phosphospecific protein kinase B (PKB/Akt) and focal adhesion kinase (FAK) protein stimulated by epidermal growth factor (EGF) was also detected. Apoptosis was determined by flow cytometry with a double-staining method using FITC-conjugated annexin V and PI. MDA468 cells transfected with PTEN gene could express PTEN mRNA and protein. PTEN decreased the phosphorylation level of AKT protein and down-regulated FAK protein expression in MDA468 stimulated by EGF. The apoptosis rate was 21.68%. PTEN induced breast cancer apoptosis phosphatase activity-dependently. The mechanism is possibly related with phosphoinositide 3-kinase (PI3K)/protein kinase B (PKB)/AKT signaling pathway. Those results may provide new clues on the gene therapy in breast cancer.

Objective To investigate exogenous PTEN gene transfected human breast cancer cell line MDA MB 468. MethodsUsing the lipofectamine 2000 transfection technique,wild type PTEN gene was transducted into an in vitro cultured highly metastatic breast cancer cell line MDA MB 468.After transfection, the cells were selected by G418.Then resistant clones were chosen and expanded in DMEM culture medium. RT PCR, immunohistochemical method and western blot were used to determine the expression of target genes. ResultsAn anti G418 cell clone was established and expanded in culture. The transfected PTEN gene MDA MB 468 cells showed expression of PTEN mRNA and PTEN protein. ConclusionHuman breast cancer cell line MDA MB 468 established in this study expresses consistently exogenous PTEN genes.

AIM: To study the relationship between the disturbance of nitric oxide/endothelin-1(NO/ET-1) and hepatic ischemia/reperfusion(I/R) injury as well as the regulation of NO/ET-1 system by hepatic ischemic preconditioning(IPC). METHODS: The changes of NO/ET-1 system and their relationship with hepatic I/R injury were compared between I/R group and IPC+I/R group in a rat hepatic I/R model. Two hours after reperfusion, the liver tissues were detected by RT-PCR to see whether there was inducible nitric oxide synthase (iNOS) mRNA expression. RESULTS:In the acute phase of hepatic reperfusion, the ratio of NO/ET-1 was reduced, which was due to a significant reduction of NO - 2/NO - 3 (the metabolic product of NO) and significant elevation of ET-1 in the blood plasma. The content of ALT, AST, LDH and TNF-? in blood plasma, and of MDA in liver tissue were increased but ATP in liver tissue was reduced, the hepatic damage was deteriorated. The protection of the hepatic IPC was concerned with the elevation of the ratio of NO/ET-1 caused by the elevation of NO - 2/NO - 3, and reduction of ET-1 as well. There was no iNOS mRNA detected in the liver tissues.CONCLUSION: Hepatic I/R injury is related to the disturbance of NO/ET-1. The protection of the hepatic IPC in the acute phase might be conducted by its regulation of NO/ET-1 system. The cNOS rather than the iNOS generated the NO in this situation.

Objective To study the inhibit effect of survivin short hairpin RNA on the survivin mRNA and protein expression of gallbladder carcinoma cells.Methods Human gallbladder carcinoma cells(GBC-SD) were transfected with recombinant plasmid.RT-PCR and Western blot were used to analyze the changes in(expression) levels of survivinmRNA and protein.Results The size of the PCR product was 350bp.DNA(sequencing) showed that the sequence of recombinant vector pshRNA-survivin was successfully constructed and suppressed the expression of(GBC-SD) survivinmRNA.Conclusions The recombinant plasmid constructed can inhibit the expression of survivinmRNA in transfected cells.This provides a new method and material for the biological therapy of cancer.

The relationship between the hepatic ischemia/reperfusion (I/R) injury and the balance of nitric oxide/endothelins (NO/ET) was studied. The changes of the ratio of NO/ET and the hepatic injury were observed in a rat hepatic I/R model pretreated with several tool drugs. In the acute phase of hepatic I/R injury, the ratio of plasma NO/ET was reduced from 1.58 +/- 0.20 to 0.29 +/- 0.05 (P < 0.01) and the hepatic damage deteriorated. NO donor L-Arg and ET receptor antagonist TAK-044 could alleviate the hepatic I/R injury to some degree, whereas NO synthase inhibitor L-NAME aggravated the damage. It was concluded that the hepatic I/R injury might be related with the disturbance of the NO/ET balance. Regulation of this balance might have an effect on the I/R injury.
Arginine ; Endothelins/*blood ; Liver/*blood supply ; NG-Nitroarginine Methyl Ester ; Nitric Oxide/*blood ; Receptors, Endothelin/antagonists & inhibitors ; Reperfusion Injury/*blood

Whether M3 cholinergic receptor signal transduction pathway is involved in regulation of the activation of NF-kappaB and the expression of chemokine MOB-1, MCP-lgenes in pancreatic acinar cells was investigated. Rat pancreatic acinar cells were isolated, cultured and treated with carbachol, atropine and PDTC in vitro. The MOB-1 and MCP-1 mRNA expression was detected by using RT-PCR. The activation of NF-kappaB was monitored by using electrophoretic mobility shift assay. The results showed that as compared with control group, M3 cholinergic receptor agonist (10(-3) mol/L, 10(-4) mol/L carbachol) could induce a concentration-dependent and time-dependent increase in the expression of MOB-1, MCP-1 mRNA in pancreatic acinar cells. After treatment with 10(-3) mol/L carbachol for 2 h, the expression of MOB-1, MCP-1 mRNA was strongest. The activity of NF-kappaB in pancreatic acinar cells was significantly increased (P<0.01) after treated with M3 cholinergic receptor agonist (10(-3) mol/L carbachol) in vitro for 30 min. Either M3 cholinergic receptor antagonist (10(-5) mol/L atropine) or NF-kappaB inhibitor (10(-2) mol/L PDTC) could obviously inhibit the activation of NF-kappaB and the chemokine MOB-1, MCP-1 mRNA expression induced by carbachol (P<0.05). This inhibitory effect was significantly increased by atropine plus PDTC (P<0.01). The results of these studies indicated that M3 cholinergic receptor signal transduction pathway was likely involved in regulation of the expression of chemokine MOB-1 and MCP-lgenes in pancreatic acinar cells in vitro through the activation of NF-kappaB.