Objective To observe the influence of losartan on rats′blood pressure and myocardium.Methods To randomly divide 24 4-year-old spontaneously hypertensive male rats into 3 groups.Group A is given normal saline(NS);group B is given losartan 20 mg/kg in 4-week-old;group C is given the same dose in 1 2-week-old;To take another 4-week-old male wistar rats to form group D,feed NS served as normal comparison.Each group was given NS or losartan one time each day through intragastric administration for 4 weeks.Follow up 24 weeks after stopping the medicine.Rats’SBP were measured through tail cuff method respectively when at 4,8,12,16,20 and 24 weeks’ time.After 24 weeks,cut off the head to take the sample,the RVW/BW,HW/BW,LVW/BW were measured through weighing method.To observe the morphologic change of the cardiac muscles and measure the area of myocardium cell cross section through HE stain method.Ra-dioimmunoassay was used to measure the AngⅡand Aldo level of the blood plasma and myocardium.Results At the age of eight-week,the SBP of group A and C are higher than those of group D(P<0.05),group B significantly lower than group A.After stopping the medicine, SBP of group B is slowly increasing and still lower than group A(P<0.05)as well as group C (P<0.05)until the 24th week.At the 16th week,SBP of group C is significantly lower than that of group A(P<0.05)but higher than group D,but quickly increases after stopping the medicine till the 24th week and there is no significant difference between group A and C(P>0.05).About LVW/BW,HW/BW and the ar-ea of myocardium cell cross section,group A increased significantly in comparison with group D (P<0.05),group B and C decreased in comparison with group A(P<0.05),group B decreased significantly in comparison with group C(P<0.05).Through the observation of the area of myocardial cell cross section under the microscopy,group A increased significantly in comparison with group D about myocardial cell fattened and the gap between cells are widened,the fibers of mesenchyma proliferates.group B and D are similar,group C is between group B and A.The level of AngⅡand Aldo in plasma and myocardium of group A increased significantly in comparison with group D (P<0.05),group B decreased significantly in comparison with group A and C(P<0.05).Conclusion To use losartan for a short time in trea-ting rats with prehypertension can get the after-effect of reducing blood pressure and anti-ventricular-remodeling.

Objective To evaluate the choice of therapeutic methods and MRI diagnosis of acoustic neuroma.Methods The MRIimaging and clinical materials of 60 patients with acoustic neuroma were analyzed,48 cases were underwent an operation,12 cases weretreated conservatively or gamma knife treatment,follow-up ranged from 1 to 4 years.Results There were 62 tumors round the internalauditory canal.There were 58 cases with single acoustic neuroma and two cases with couples acoustic neuroma.28 tumors demonstratedhypointense and 30 tumors demonstrated hypo-and isointense signals on T_1 weighted image.38 tumors demonstrated hyperintense and 24tumors demonstrated hyper-and isointense signals on T_2 weighted image.The Ⅶ,Ⅷ nerves affected side were thickened than that ofopposite side in 32 patients.After Gd-DTPA administration 24 tumors were homogeneously enhanced,26 tumors were inhomogeneously or circularly enhanced in 50 acoustic neuromas of 48 cases.Operation was still the main election for acoustic neuroma.Conclusion MRI is an effective method in the diagnosis of acoustic neuroma,and providing advice for clinics in making therapeutic programs.

Objective to investigate the role of Pim‐3 and NF‐κB in the development and progression of infiltrating ductal carcinoma of the breast .Methods Here ,we used immunohistochemistry to detect expression of Pim‐3 and NF‐κB in 75 samples of infiltrating ductal breast carcinoma ,21 samples of intraductal breast carcinoma and 30 normal breast tissues .The relationship of their expression ,as well as their correlation with clinicopathological features and patient survival were assessed .Results In con‐trast ,both Pim‐3 and NF‐κB were more commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue .In the infiltrating ductal carcinoma ,the positive expression rate of Pim‐3 was 77 .3% ,and that of NF‐κB was 68 .0% ;in duc‐tal carcinoma of the breast ,the positive expression rate of Pim‐3 was 52 .4% ,and that of NF‐κB was 42 .9% ;in the normal breast tissue ,the positive expression rate of Pim‐3 was 23 .3% ,and that of NF‐κB was 16 .7% ;the positive expression rate of Pim‐3 was correlated with tumor size ,histological grade ,and clinicopathological stage ;and that of NF‐κB was correlated with tumor size ,histo‐logical grade ,lymph node metastasis of breast cancer .Spearman rank correlation analysis revealed a positive correlation between Pim‐3 expression and NF‐κB expression in infiltrating breast cancer (r=0 .243) .Conclusion Our results demonstrate that Pim‐3 and NF‐κB play a role in the initiation and development of breast cancer ,thus ,these proteins may serve as useful diagnostic and prognostic markers of invasive breast cancer .

AIM: To construct a mouse IFN-? expression vector and observe the antitumor effects of mouse peritoneal macrophages transfected with IFN-? in vivo and in vitro. METHODS: The IFN-? mRNA was amplified by RT-PCR. The open reading frame of mouse IFN-? gene was recombinanted with eukaryotic expression vector pcDNA3.1 through subcloning. Mouse peritoneal macrophages were transfected with recombinant vector pcDNA3.1-IFN-?. The expression of INF-? mRNA was measured by RT-PCR. Another group of peritoneal macrophages were cultured with the culture medium from pcDNA3.1-IFN-? transfecting groups, and its antitumor effect was measured by MTT. pcDNA3.1-IFN-? plasmid was peritoneally injected inte mouse with tumor. The appearance of ascites of pcDNA3.1-IFN-? plasmid injected mice and survival time were observed. RESULTS: The mouse IFN-? expression vector pcDNA3.1-IFN-? was constructed. The sequence was demonstrated to be the same as on GenBank. The recombinant vector was introduced into mouse peritoneal macrophages. IFN-? mRNA was detected by RT-PCR. The supernatant from cultured macrophages transfected with pcDNA3.1-IFN-? plasmid stimulated the antitumor effects of the macrophages without transfection. The appearance of ascites in pcDNA3.1-IFN-? plasmid injected mouse was delayed and survival time was longer than that in other groups. CONCLUSION: We have successfully constructed the mouse IFN-? expression vector pcDNA3.1-IFN-?. Mouse peritoneal macrophages transfected with pcDNA3.1-IFN-? have antitumor effects in vivo and in vitro.

The differentially expressed genes between esophageal squamous cell carcinoma (ESCC) with or without lymphatic metastasis were investigated by gene chip, and the lymphatic metastasis-associated genes were screened out. Expression array was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human ESCC. The lymphatic metastasis-associated genes were screened by bioinformatics between ESCC with or without lymphatic metastasis. The results showed that 43 (4.85 %) genes significantly differed between the ESCC with and without lymphatic metastasis (P<0.05), of which 18 (2.03 %)were upregulated and 25 (2.82 %) down-regulated. The up-regulated genes were involved in cell adhesion molecules and cell membrane receptors and the down-regulated genes were mostly cell cycle regulators and intracellular signaling molecules. It was suggested that lymphatic metastasis-associated genes were screened by gene chip, which was helpful to understand the molecular mechanism of ESCC lymphatic metastasis and lymphatic metastasis-associated genes might be used as diagnostic markers and therapeutic targets for lymphatic metastasis.

Objective To investigate the differentially expressed genes of primary esophageal squamous cell carci-noma and of normal esophageal mucosa. Methods LCM-GMA-cDNA microarray was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human esophageal squamous cell carcinoma (ESCC). After high-stringent washing, the cDNA microarray was scanned for the fluores-cent signals. Results Among the 886 target genes, 34 genes had significant difference in Ⅰ / Ⅱ and Ⅲ/Ⅳ group. Cell cycle regulators possibly promoting the growth of tumor cells were highly expressed in the early stages of ESCC, whereas adhesion molecules and extracellular matrix-related molecules possibly promoting invasiveness increased in the later stages. Conclusion More than one gene contributed to esophageal cancer. The profiles of gene expression will bring us chance to understanding the molecular mechanism of tumor progression and to support clinical treat-ment.

AIM: To prepare the vaccine of DCs(pcDNA3-hCEA) and observ the immunity effect of the DCs(pcDNA3-hCEA) inoculating on CT26(hCEA +) loaded in BALB/c mice. METHODS: DCs were generated from bone marrow in the presence of rmGM-CSF and rmIL-4. A new recombinant plasmid, pcDNA3-hCEA, reformed with inserting a 2.4 kb human CEA cDNA into pcDNA3. DC vaccine was prepared by transfection with pcDNA3-hCEA using lipofectamine. CEA mRNA expressed in DCs(pcDNA3-hCEA) was confirmed by RT-PCR, CEA expression level was detected with RIA method, and CEA specific CTL was induced in vitro . After vaccination of DCs(pcDNA3-hCEA), the survival time of the BALB/c mice challenged with critical loading CT26(hCEA +) was observed. RESULTS: G418 test showed that about 14% DCs were transfected with pcDNA3-hCEA. And CEA mRNA and protein could be detected respectively by RT-PCR and RIA in the genetically modified DCs. Furthermore, the DCs coud be targeted by specific CTL, the survival time of the mice challenged with CT26(hCEA +) was prolonged 1-4 weeks. CONCLUSION: These results demonstrate that specific antitumor immune responses could be induced efficiently by vaccination of DCs(pcDNA3-hCEA), which is transfected eukaryotic expression vector encoding tumor antigen gene.

The differentially expressed genes between esophageal squamous cell carcinoma (ESCC)with or without lymphatic metastasis were investigated by gene chip, and the lymphatic metastasisassociated genes were screened out. Expression array was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human ESCC. The lymphatic metastasis-associated genes were screened by bioinformatics between ESCC with or without lymphatic metastasis. The results showed that 43 (4.85%) genes significantly differed between the ESCC with and without lymphatic metastasis (P＜0.05), of which 18(2.03%)were upregulated and 25 (2.82 %) down-regulated. The up-regulated genes were involved in cell adhesion molecules and cell membrane receptors and the down-regulated genes were mostly cell cycle regulators and intracellular signaling molecules. It was suggested that lymphatic metastasis-associated genes were screened by gene chip, which was helpful to understand the molecular mechanism of ESCC lymphatic metastasis and lymphatic metastasis-associated genes might be used as diagnostic markers and therapeutic targets for lymphatic metastasis.

Objective To investigate the etfect of microenvironment simulated by colon carcinoma homogenate supernatant on the differentiation and development of human dendritic ceils (DCs), and to investigate the function of vascular endothelial growth factor A (VEGF-A) during this process . Methods Fresh colon carcinoma and peri-cancer tissues were collected to prepare homogenate supernatant. The pe-ripberal blood mononuclear cells were isolated and cultured with 1640 medium including rhGM-CSF and rhIL-4. Then the colon carcinoma homogenate supernatant, peri--carcinoma homogenate supernatant and VEGF-A were added to the cultures at day 2. Antigen of colon carcinoma cell line SW620 was added at day 4 and lipopolysaccharide (LPS) was added at day 6. DCs were collected at day 8 for further study. The con-tent of VEGF-A was tested by ELISA. The morphology and the immunopbenotype of DCs were checked by microscope and flow cytometry, respectively. The expression of CDIa was tested by RT-PCR, and the prolif-eration and killing rate of T cell was measured by CCK-8. Results The content of VEGF-A in the homoge-nate supernatant of colon carcinoma was significantly higher than that of the peri-carcinoma (P ＜ 0. 05). Compared with normal DCs, the cell morphology of colon carcinoma homogenate aupernatant group was in-hibited, and the cell number was decreased. Besides, the positive expression rate of DC surface markers de-creased (P ＜ 0.01). The capacity of mixed lymphocyte reaction (MLR) and killing capacity of T cells de-creased(P ＜0.01). However, there was almost no difference between VEGF-A group and normal DCs on the cell morphology and cell number, and VEGF-A had no obvious inhibition on the expression of DCs sur-face markers (P ＞ 0.05). But VEGF-A group had significantly inhibitory effect on the MLR and T cells kill-ing. Conclusion The tumor microenvironment simulated by the colon carcinoma homogenate supernatant obviously has inhibitory effect on the differentiation and function of DCs, and VEGF-A has the inhibitory effect on DC function, but the inhibitory effect is not through the inhibition of the expression of DC costimu-lators.

Objective To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell of mice. Methods Mouse brain microvascular endothelial cell bEnd. 3 was cultured and identified for preparation endothelial cell bEnd. 3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R2) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd. 3 antigen 4 times in 4 weeks (bEnd. 3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD3+ CD+8 surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot. Results The expression of VEGF-R2 and integrin αV gene in bEnd. 3 cells were expressed highly.After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd. 3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11± 0.13) cm3 versus (3.38 ±0.34) cm3]. The CTL response of spleen iymphocytes in vitro showed that bEnd. 3-DC cells could kill bEnd. 3 cells, the special lysis rate was more than 60% . The percentage of CD+3 CD+8 spleen lymphocytes in bEnd. 3-DC group[(38.6 ± 0.7) %] was higher than those in other groups (P ＜ 0.05). The titer of serum antibody of Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd. 3 cell in bEnd. 3-DC group. Western blot analysis revealed that there were specific bands at 220 000 (VEGF-R2).Conclusions bEnd. 3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humorai immunity are induced by bEnd. 3-DC antigen which maybe have some antigens in bEnd. 3 cells that reacts with endothelial cell proliferation-related antigens.