Objective To investigate the expression of snrvivin,an anti-apoptosis gene,in breast cancer tissues and its relationship with clinical parameters of breast cancer patients.To investigate the role of survivin in oncogenesis of breast cancer and its relation with prognosis of breast cancer.Methods Western blot was conducted to detect survivin expression in cancerous tissues and adjacent tissues of 50 breast cancer patients.20 normal breast tissue samples were used as control.Results Survivin was expressed in 38 of the 50 breast cancer tissues (76.0%)and in 11 of 50 breast adjacent normal tissues(22.0%)while no expression was found in normal tissues.The positive protein expression of survivin was related to axillary lymph node metastasis of breast cancer while it was not related to expression of PR,ER,Her-2 or ages of patients.Conclusion Survivin may play a role in breast tumorigenesis and is an important predictive parameter in breast cancer patients.

Objective Study on the intracellular calcium and growth suppression of human breast cancer cells exposed by high intensity ultrasound (HIU). Methods Exposed human breast cancer cells MDA-MB-231 and MCF-7 in vitro with HIU (50 W/cm2). Examined the intracellular calcium from exposed cells with Fura-2 fluorescence probe. The cell viabilities were measured by MTT assay. The rate of cell apoptosis and distribution of cell cycle were detected on flow cytometry. Results The lever of intracellular calcium went up in pace with exposed time of HIU, MCF-7 cells were (572±20.1), (670±18.9), (815± 16.3) nmol/L (F = 663.65, P＜0.001), MDA-MB-231 cell was (582±16.3), (687±19.7), (843± 14.8) nmol/L (F = 863.06, P＜0.001), and the distribution of cell cycle waved to G0-G1, the ratios of G0-G1 in MCF-7 and MDA-MB-231 were (60.5±5.5)%, (66.3±7.0)%, (74.5±8.2)% (F=8.17, P = 0.002) and (58.5± 6.3) %, (66.1±6.3)%, (71.2±7.9) % (F=7.51, F= 0.003). Apoptotic rate upgraded gradually, the apoptotic rates of MCF-7 and MDA-MB-231 were (7.3±1.7)%, (13.2±3.5) %, (19.3±3.7)% (F= 18.73, P＜0.001) and (6.3±1.8)%, (11.4±2.31)%, (16.4±3.3)% (F = 19.26, P＜0.001). Under MTT assay, the rate of cell growth suppression increased significantly, the rates of cell growth suppression in MCF-7 and MDA-MB-231 were (9.2±2.2) %, (24.3±3.9)%, (48.6±5.5)%(F=117.16, P ＜0.001) and (9.0±1.7)%, (22.3±3.5) %, (416± 6.4)% (F =71.25, P＜0.001). Conclusion HIU enhanced the intracellular calcium of human breast cancer cells within given time and promoted the distribution of cell cycle to G0-G1. The rate of cell apoptosis and the cell's death rate increased evidently.

Objective To explore the influence of different dose of Helicobacter pylori on the expression of transforming growth factorβ(TGF-β1) and B7-H1 in the infected gastric mucosal epithelial cell and the bacterial factors which influence the expression of TGF-β1.To confirm that H.pylorican induce the expression of TGF-β1 and B7-H1 to inhibit the host immune function in gastric mucosal epithelial cell.Methods (1) We investigated the expression of TGF-β1 of human gastric mucosal epithelial cells infected with different concentration(1.0 × 109 CFU/ml,4.0 × 109 CFU/ml,8.0 × 109 C FU/ml) of H.pylori(NCTC 11637) in different time-point(0 h,0.5 h,1 h,1.5 h,2 h,4 h,8 h,12 h),and compared with the expression of TGF-β1 between the deactivated H.pylori group and activated H.pylori group.The blank group is the gastric mucosa epithelial cells which does not infect H.pylori.To detect expression of TGF-β1 in infected cell culture supernatant by enzyme-linked immunosorbent assay(ELISA) and the expression of B7-H1 mRNA by in situ hybridization.(2)At the same time,the middle concentration of deactivated H.pyloriand in vitro gastric mucosal cells were incubated for 2 h and 12 h,to detect expression of TGF-β1 in the cells and cell culture supernatant.(3)In vitro gastric mucosal cells were incubated with H.pylori bacterial supernatant and sedimentation by ultrasonic crushing and centrifugation and with H.pyloribacterial supernatant and sedimentation after boiling respectively,to detect expression of TGF-β1 in the cells and cell culture supernatant after 2 h,12 h.Results (1)Compared to the control group,the expression of TGF-β1 was significantly increased after stimulation with different concentration of activated H.pylori in different time-point(P ＜0.05).The expression of TGF-β1 secretion group has a similar dynamic trend,and the highest expression is the middle concentration group(P ＜0.05).(2)There was no difference between the middle concentration of deactivated H.pylori group and the same concentration of activated H.pylorigroup(P ＞ 0.05).(3) The expression of TGF-β1 in the H.pylori bacterial supernatant group was significantly increased higher than the blank group and the H.pylori bacterial sedimentation group(P ＜0.05),and the H.pylori bacterial supernatant group after boiling was significantly lower than the H.pylori bacterial supernatant group(P ＜ 0.05),but there was no difference between H.pylori sedimentation group after boiling and not boiling(P ＞ 0.05).The B7-H1 expression of different concentration groups which the H.pylori and gastric mucosal epithelial cells cocultured 12 h are higher than the blank group(P ＜ 0.05) by in situ hybridization,and the middle concentration group is the highest expression.TGF-β1 and B7-H1 mRNA are positively correlated(r,=0.628,P ＜0.01).Conclusion H.pylori can induce the gastric mucosal epithelial cells to express the TGF-β1,the factor was the soluble protein in the H.pylori thalline.At the same time,H.pylorican induce the B7-H1 expression increased.In gastric mucosal epithelial cells,TGF-β1 and B7-H1 mRNA are positively correlated.So H.pylori can inhibit the host immune response and participate the process of immune escape by increased the expressions of TGF-β1 and B7-H1.

Objective To explore the influence of Helicobacter pylori (Hp) on the proliferation, apoptosis and expression of p27kipl protein of gastric epithelial cells in vitro. Methods SC, C-7901 cell pro-liferation was examined by flow cytometry after incubation with different concentration of NCTC11637. The effect of NCTC11637 on cell apoptosis was also evaluated by flow cytometryo And the expression of p27kipl of gastric epithelial cells was detected by immunohistochemical analysis and in situ hybridization, respectively. Results Cell proliferation was enhanced when co-incubated with the Hp in low concentration (3.4 x 104 to 1.9 x 105 CFU/ml) but inhibited in higher concentration (≥4.8×106CFU/ml). However, cell apoptosis was increase when co-incubated with the Hp in high concentration (≥9.6 ×105 CFU/ml) showing concen-tration dependent picture. In addition, the co-incubation of SGC-7901 with Hp led to decrease of the expres-sion of p27kipl protein but not mRNA in a Hp concentration dependent way. Conclusion Hp could effect the gastric epithelial cells apoptosis and proliferation directly and influence the expression of p27kips protein which might facilitate gastric carcinoma.

In this study,the effects of pirrolidine dithiocarbamate (PDTC) plus leflunomide (Lef) models were investigated.NIH mice and Wistar rats served as donors and recipients respectively.Mouse-to-rat cardiac xenotransplantation was performed.The recipients were divided into 5 groups:group A (the control group),group B (PDTC group),group C (PDTC plus CsA group),group D (PDTC plus Lef group) and group E (PDTC plus Lef and CsA group).The expressions of IKKα/β,by immunohistochemistry and Western blot as well as electrophoretic mobility shift assay (EMSA).The median survival time of cardiac xenografts in the control group,PDTC group,PDTC plus CsA group,PDTC plus Lef group and PDTC plus Lef and CsA group was (2.17±0.41),(2.33±0.52),(4.67±1.21),(7.00±1.79) and (9.00±1.41) days respectively.The survival time of xenografts in the PDTC plus Lef and CsA group was significantly longer than that in other four groups (P<0.05 for all),that in the PDTC plus Lef group longer than that in the control group,PDTC group and PDTC plus CsA group (P<0.05 for all),that in PDTC plus CsA group longer than the control group and PDTC binding activity were notably increased in mouse-to-rat cardiac xenografts.The expressions were decreased in the control group,PDTC group,PDTC plus CsA group,PDTC plus Lef and PDTC plus Lef and CsA group in turn.It was concluded that PDTC plus Lef and CsA can significantly suppress prolonging the survival of the cardiac xenografts.

Objective To investigate the protective effect of heme oxygenase-1 for xeno-heart transplantation and the possible mechanism.Methods Cobalt protoporphyrin (CoPP), HO-1 inducer, and zinc protoporphyrin (ZnPP), HO-1 inhibitor, were used to intervene donors and recipients on the model of NIH-Wistar cardiac transplants respectively and simultaneously some of recipients were treated with immunesuppressor cyclosporine A (CsA), and control group and CsA treated group were set up respectively. The expression of HO-1 protein, HO-1 mRNA, caspase-3 and STAT-3 in transplanted heart tissue was detected by immunohistochemistry, RT-PCR and Western Blot. At the same time, HO-1 enzymatic activity was examined in heart tissue, and the cardiac cell apoptosis was examined by means of TUNEL. The differences among various factors were compared.Results The survival time of cardiac transplants in CoPP group was longer than that in control and ZnPP groups (P

Objective To review the mechanisms of breast cancer escaping from host immune surveillance. Methods The current literatures on the mechanisms of breast cancer cells escaping from host immune surveillance were reviewed in the following aspects: alterations of MHC-Ⅰmolecule phenotype, deficiency of costimulatory molecules, apoptosis of T-lymphocytes induced by breast cancer cells presenting Fas ligand (FasL) and TNF-related apoptosis-inducing ligand (TRAIL) and host immune tolerance induced by tumor cells. Results Loss of classical antigen-presenting human leukocyte antigen (HLA) class-Ⅰmolecules, expression of non-classical HLA class-ⅠmoleculesHLA-G, loss of costimulatory molecule B7, apoptosis of T-lymphocytes induced by tumor cells presenting FasL and TRAIL and antigen-inducing host immune tolerance were related to breast cancer escaping from immune surveillance. Conclusion Breast cancer cells escape from host immune surveillance by altering MHC-Ⅰmolecules, lacking of costimulatory molecules, inducing of apoptosis of T cells and host immune tolerance. But the factors resulting in breast cancer escaping from host immune surveillance are not yet clear and should be further studied.

OBJECTIVESTo observe the expression of somatostatin receptor subtype 2 (SSTR2) mRNA, and investigate the relationship between the expression of SSTR2 mRNA and the expressions of estrogen and progesterone receptors (ERs and PRs) in benign and malignant breast tissues.

METHODSSamples from a total of 23 breast carcinomas, 16 mammary hyperplasias, and 9 mammary fibroadenomas were analyzed. SSTR2 mRNA expression was examined by in situ hybridization using multiphase oligoprobes. ER and PR expressions were detected by immunohistochemical staining. A computerized image analysis system was utilized to estimate the relative content of SSTR2 mRNA.

RESULTSThe rate of expression (87.0%) and relative content (0.47) of SSTR2 mRNA in breast cancer were higher than those in benign breast tissue (64%, 0.26) (P < 0.05). SSTR2 mRNA expression was closely correlated with ER and PR expressions in breast cancer (P < 0.05). SSTR2 mRNA was also positively correlated with ER expression in benign breast tissues.

CONCLUSIONSSSTR2 mRNA expression is higher or in benign breast tissues than in malignant ones. There is a significant positive correlation between SSTR2 mRNA and ER and PR expressions. Combined antiestrogen and somatostatin analogue in treatment of ER-positive breast cancers should be further investigated.

Breast Diseases ; metabolism ; Breast Neoplasms ; chemistry ; Humans ; Immunohistochemistry ; In Situ Hybridization ; RNA, Messenger ; analysis ; Receptors, Estrogen ; analysis ; Receptors, Progesterone ; analysis ; Receptors, Somatostatin ; genetics

Objective By reproducing mouse to rat heart xenograft,to determine the effect of leflunomide on NF-?B P65 in the transplanted heart.Methods The recipient rats were randomly divided into 4 groups:group A(n=6)receiving no immunosuppressant as control;group B(n=6)cyclosporine(CsA);group C(n=6)leflunomide(Lef),group D(n=6)Lef plus CsA.The expressions of NF-?B P65 protein and NF-?B P65 DNA binding activity in transplanted heart tissues were determined by immunohistochemistry,Western blot and electrophoretic mobility shift assay(EMSA).Results Administration of CsA alone had no appreciable effect on the survival time of xenografts [(2.50?1.05)d] and could not suppress the protein expression of NF-?B P65 [(263.09?28.81),IOD] and NF-?B P65 DNA binding activity [(227.49?14.83),IOD] in the tissues of xenografts.Administration of Lef alone could significantly prolong the survival time of xenografts [(4.17?1.33)d] and could suppress the protein expression of NF-?B P65 [(173.48?5.85),IOD] and NF-?B P65 DNA binding activity [(109.96?12.46),IOD](P

Objective To study the liver injury and effects of aescin on liver in rats with acute pancreatitis. Methods The rats were divided into 3 groups (control group, AP group and aescin group). The serum alanine aminotransferase (ALT), serum lactate dehydrogenase (LDH), hepatic cellular energy charge (EC) and adenosine triphosphate (ATP) were detected. The pathologic changes in pancreas and liver were also observed. Results The serum levels of ALT and LDH in aescin group were significantly lower than those of the AP group. The EC and ATP levels were significantly higher in aescin group than that of the AP group. Conclusion Introvenous injection of aescin can alleviate the liver injury in rats with acute pancreatitis.