ObjectiveTo evaluate mammaglobin (hMAM) mRNA expression as a tumor marker for micrometastatic carcinoma cells in the peripheral blood of patients with breast cancer (BC). MethodsTotal RNA extracted from a breast carcinoma cell line SKBR3 was subjected to analysis of hMAM mRNA expression by RT-PCR. The sensitivity of FQ-PCR to detect a SKBR3 cell at a level of 10 7 was established. Peripheral blood of 63 BC patients was examined by this method. ResultsBlood samples were positive in 19 out of 63 (30%) patients with breast carcinomas. None of the patients with other cancer (25 cases) or benign breast disease (13 cases) was positive and only 1 out of 31 healthy volunteers was found with detectable hMAM mRNA expression in the peripheral blood. ConclusionFluorescence quantitative (FQ) PCR combined with nest PCR was a sensitive method in detecting micrometastases of breast cancer.

ObjectiveTo evaluate portal vein chemotherapy for the treatment of hepatocellular carcinoma with portal vein tumor thrombi(PVTT).MethodsFourty five HCC patients with tumor thrombi in the main portal vein or the first branches undergoing surgical resection were divided into two groups receiving postoperative intraarterial and intraportal chemotherapy (21 cases), and intraarterial chemotherapy only (24 cases).ResultsThe survival rates of 6,12,18 months was 86%, 76% and 48% in the treatment group compared with 75%, 58% and 33% in the control group respectively. There were significant difference between the two groups.Conclusion Hepatectomy for HCC with removal of tumor thrombi and transcatheter hepatic arterial and portal vein chemotherapy is an effective treatment for HCC with PVTT.

Purpose:To explore whether expression of BRCA1 is altered during the development of sporadic ovarian epithelial cancer. To investigate the relationship between BRCA1 expression and clinical pathological parameters. Methods:The expression of mRNA and protein of BRCA1 were determined in 50 ovarian cancers, 11 benign ovarian tumors and 10 normal ovary tissues by means of RT PCR and S P immunohistochemical method. Results:BRCA1 mRNA relative expression levels were 0.6688?0.2232?0.9264?0.1398 and 1.0440?0.2182, respectively, and protein expression rates were 30.0%, 90.9%, and 100% in ovarian cancers, benign tumors and normal ovary tissues separately. The expression levels were significantly lower in ovarian cancers than that in the latter two. In addition, the level of BRCA1 expression decreased with pathological grade increasing; and also BRCA1 expression reduced, even deleted, in cancers with pelvic lymph node metastasis. Conclusions:Down regulation of BRCA1 expression at the levels of transcription and translation plays an important role in the occurrence, and progression of sporadic ovarian cancer. Reduction of BRCA1 expression is closely related to tumor pathological grade and probably implies a poor prognosis.

Objective: To evaluate the efficacy of intraoperative hyperthermic peritoneal perfusion (CHPP) on advanced gastric carcinoma. Methods: Sixty patients with advanced gastric carcinoma were divided into the control group and the treatment group. All patients underwent radical gastrectomy and D2 node dissection. Patients in the treatment group received CHPP when surgical resection was completed. Patients in the control group underwent resection of gastric carcinoma without CHPP. Chemotherapy was administered with FOLFOX4 regimen intravenously for 12 cycles in both groups at 4 weeks after surgery. The serum Carcinoembryonic antigen (CEA) and CA19-9 were measured in patients with advanced gastric cancer before and after resection of tumor. Survival and recurrence in both groups were analyzed and compared. Results: The mean levels of the expression of CEA and CA19-9 in the peripheral blood of the 60 patients were significantly higher than the upper limits of normal (55.89±22.25μg/L vs 0～5μg/L; 125.35±61.78 U/mL vs 0～39U/mL P< 0.01). There were no significant differences in the mean levels of the expression of CEA and CA19-9 in the peripheral blood between the treatment group and the control group (54.67±22.95μg/L vs 56.09±22.15μg/L; 126.16±62.45 U/mL vs 123.35±60.88 U/mL,P>0.05). The serum CEA and CA19-9 levels were significantly decreased at 7 days after treatment in the treatment group (7.58±3.21 μg/L, 31.35±13.47 U/mL, P<0.01). The levels of these two tumor markers were decreased unremarkably at 7 days after treatment in the control group (37. 68±20.59μg/L, 98.23±36.28 U/mL, P>0.05). The serum CEA and CA19-9 levels were decreased significantly in both groups at 30 days after surgery (P<0.05). One-year survival and recurrence rates were 83.3% and 10% in the treatment group and 80% and 13.3% in the control group, with no significant differences between the two groups (P>0.05). Three-year survival and recurrence rates were 63.3% and 20% in the treatment group and 40% and 40% in the control group, with a significant difference between the two groups (P<0.05). Conclusion: Surgical resection combined with CHPP can significantly decrease the serum CEA and CA19-9 levels. Intraoperative CHPP for patients with advanced gastric carcinoma is helpful for preventing peritoneal metastasis and recurrence and can prolong survival time.

Objective:To study the distribution pattern of CIK cells re-infused by different manner.Methods:Isotope 32P-? dATP and fluorescence dye CM-DiI were used individually to label CIK cells. CIK cells labeled by the two methods in vitro were inoculated to nude mice by intraperitoneal injection or tail vein injection. Radioactivity quantitative measurement and fluorescence microscopy were used to analysis dynamic distribution of CIK cells among organs of mice.Results:The CIK cells were quickly distributed to organs such as liver, spleen, kidney, lung, stomach and intestine after inoculation into nude mice. Among those organs, the liver, spleen and kidney showed highest distribution concentration of CIK cells. Early stage after infusion, concentration of CIK cells in lung above all reached peak via tail vein, and by means of intraperitoneal injection, distribution of CIK cells in intraperitoneal organs firstly got to max. CIK cells remained alive in liver and spleen for more than 2 weeks.Conclusion:The extensive distribution pattern of CIK cells among organs shows that CIK cells can be used as drugs against various malignant tumors in organism. Infusion of CIK cells via blood vessel maybe suit for tumor of organs with rich blood supply, and application by means of body-cavity way should suit for malignant effusions and limited lesion in it.

Objective:For developing the study of MUC1/Y vaccine,constructed th eukaryotic expression vectors expressing MUC1/Y.Used it to modify the DC inducint CTL in order to treat the gastrointestinal cancers.Methods:Obtained the MCU1/Y cDNA full length cloned it into pIRES2 EGFP that expressing green fluorescence protein and pcDAN3 1+ respectively.Selected 8 cases of gastrointestinal cancer whose HLA phenotype was A2,inducing DC using rhIL 4 combined with GM CSF in vitro.Transfected DC with pcDAN3.1 MUC1/Y then co cultured DC with T cell to induce CTL (named as T pcDAN3 1 MUC1/Y).At the same time,used pIRES2 EGFP MUC1/Y to detect transfection efficiency.SW620 (HLA A2+,MUC1/Y+),the gastric cancer cell line was used as specific target cell;Lovo(HLA A2 ,MUC1/Y+)and Raji(HLA A2 ,MUC1/Y )were used as unspecific target cells.The cytolytic of specific CTL was measured by LDH releasing assay.The apoptosis of target cells were detected by ANNEXIN V FITC kit.The ability of IFN ? releasing of T cells was measured by ELISA.Results:The transfection efficiency of plasmid was about 8%.There was significant difference between the cytolytic activity of T pcDNA3 1 MUC1/Y to SW620,Lovo and Raji.The cytolytic activity was about (42 8?6 15),(27 26?1 96)% and (22 73?2 15)% respectively.Compared with T IL 2)CTL induced by PBMC stimulated of IL 2(100 U/ml)) and T pcDNA3 1(CTL induced by DC transfected by pcDNA3 1+),the cytolytic activity of T pcDNA3 1 MUC1/Y against SW620 cell line showed a significant increase.The results of ANNEXIN V-FITC experimences showed that T pcDNA3 1 MUC1/Y could induce apoptosis of specific target cell.Conclusion:The expressing of MUC1/Y mRNA has important sense in gastrointestinal cancer.Constructed eukaryotic vector pIRES2 EGFP MUC1/Y that contains full length MUC1/Y cDNA could be used to study the transfection efficiency and select the positive clone;pcDAN3 1 MYC1/Y could induce powerful CTL immunoresponse.

Objective: To explore the inhibition effects of ectogenic wild type p53 cDNA(Ad wtp53) on colorectal carcinoma cell lines with different p53 gene status and search for the role of wild type p53 tumor suppressor gene in occurrenc and progress of malignant tumor. Methods: MTT process was taken to choose optimal transfection titre. Three kinds of cell lines(p53 gene deletion, mutation and nomal) were transferred by Ad wtp53 in optimal titre. The inhibition effects of these cell lines were observed and compared. Results: The best titre is 1000 MOI and p53 gene deletion cell line (THC 8908) shew the highest sensitivity. G 1 S transition period blocking effects occurred in all cell lines and G 2 M phase regulation effects were not coincidence in three colorectal cell lines. Conclusions: Recombinant adenovirus mediated wild type p53 gene observably inhibited colorectal carcinoma cell lines growth and proliferation, blocked cell cycle in G 0 /G 1 phase and displayed obvious different actions on G 2 M phase among cell lines with different p53 status.

Objective To investigate the clinical features and prognostic factors of nasal type NIGT cell lymphoma(NKTCL).Methods Records of 61 patients with pathologically confirmed nasal type NKTCL were reviewed.Detailed clinical and laboratory data were included in univariate analysis, and statistically significant factors in univariate analysis were then included in multivariate analysis.Results In univariate analysis,Ann Arbor stage,performance status,IPI,number of extra-lymphatic site,B symptoms,LDH and β-MG level.were found to be the prognostic factors associated with time to overall survival in nasal type NKTCL.In multivariate analysis, Ann Arbor stage,performance status,LDH and β2-MG level were independent prognostic factors of overall survival.Conclusion Ann Arbor stage,performance status,LDH and β2-MG level were demonstrated as independent prognostic factors of the overall survival in nasal type NKTCL.

Objective: To analyze the circulating endothelial cells (CECs) and circulating endothelial precursors (CEPs) in peripheral blood of tumor patients. Methods: CECs and CEPs were enumerated in 57 tumor patients and 15 healthy controls by 3-color flow cytometry. The serum level of angiogenesis related cytokines(VEGF, bFGF)was determined by ELISA method. Results: In tumor patients, mean value of CECs and CEPs was (0.378?0.047)% and (0.059?0.013)% respectively. The serum level of VEGF and bFGF were (295.58?59.56) ng/L and (28.73?7.40) ng/L. The amount of CECs/CEPs positively correlated with the serum level of VEGF and bFGF. Conclusions: The CECs/CEPs and serum level of VEGF and bFGF of tumor patients was higher than those of normal controls. VEGF and bFGF may participate into the course of endothelial progenitors mobilization.

Objective:Analyze the influence of hematopoietic stem cell mobilization on circulating endothelial cells (CECs) and circulating endothelial precursors (CEPs).Methods:CECs and CEPs were enumerated in 68 tumor patients and 15 healthy controls by 3-color flow cytometry. 11 cases underwent PBSC (peripheral blood stem cell) mobilization by combination chemotherapy and G-CSF (granulocyte colony-stimulating factor).Results:CECs and CEPs in tumor patients were 0.378%?0.103% and 0.059%?0.013% respectively,which were higher than that of normal controls.CECs/CEPs in peripheral blood (PB) were increased after G-CSF mobilization.Conclusion:Hematopoietic and endothelial progenitors can be mobilized into the PB by treatment with growth factor G-CSF.