Objective To assess the effect of heat shock protein 70 of BCG (BCG HSP70) gene transfection on tumorigenicity and immunogenicity of murine lymphocytic leukemia cells (L1210). Methods BCG HSP70 gene was transfected onto the surface of murine lymphocytic leukemia cells (L1210) by lipofectamine 2000. And then the positive clone (L1210-HSP70) highly expressing HSP70 was selected as the tumor vaccine to study the tumorigenicity experiments in nude mice and syngeneic mice, the therapeutic experiments, and the immunoprotective effects. Results The expression of BCG HSP70 on the L1210 cells surface was detected, and the L1210-HSP70 cells had the same tumorigenicity as the parental L1210 cells did. Tumorigenicity experiments in syngeneic mice:In L1210-HSP70 group, tumor growth was slow or without the formation of tumor. As compared with L1210 group and L1210-neo group the mice survival time was signiifcantly prolonged, showing a marked stimulating effect on L1210 specific Th1 cells,. Tumor-bearing mice showed complete coagulation necrosis and abundant CD8+T lymphocyte inifltration (P<0.05). The tumor vaccine of L1210-HSP70 cells had the antitumor therapeutic efifcacy and immune protection effect, demonstrating that the tumor growth was signiifcantly inhibited, tumor diameter was markedly reduced and the survival time of tumor-bearing DBA/2 mice was further prolonged. Conclusions BCG HSP70 gene transfection could effectively improve the immunogenicity of tumor cells, activate speciifc T cells and enhance the anti-tumor immunity in vivo. Meanwhile, the host anti-tumor immunity could be enhanced.

Pan-retinal photocoagulation (PRP) and macular photocoagulation (MPC) are the gold standard treatments for proliferative diabetic retinopathy (DR) and diabetic macular edema. With the development of equipment and technology advancement, photocoagulation has been gradually applied in many Eye Centers all over China. However, there are still several problems such as no standardized guideline and undesirable therapeutic effects. In this article we will summarize the indications and techniques of photocoagulation, and when and how to apply drug treatments for retinal diseases; aim at improving the criterion and clinical effects of photocoagulation.

Objectives To prepare the HL-60 cell vaccine expressing heat shock protein 70 (HSP70) of Bacille calmette-Guérin (BCG), so as to study its anti-tumor effect and mechanism. Methods The whole BCG HSP70 gene was amplified from BCG genome by polymerase chain reaction (PCR) and sub-cloned into the polyclone endonuclease sites in pDisplay. The recom-binant vector of pDisplay-HSP70 was verified by sequencing. Then the HL-60 cell vaccine expressing the protein onto the cell surface was prepared by lipofectamine transfection. To detect the immunogenicity of HL-60 cells expressing HSP70, the test groups were divided into three subgroups, HL60-wt, HL60-pDisplay, and HL60-HSP70 respectively. Each group was cultured with peripheral blood T cells for 72 h, then the proliferation indices of T cells were assayed by CFSE-staining method, and IFN-γwere tested by enzyme-linked immunosorbent assay (ELISA). The HL-60 cells of different groups were cultured with peripher-al blood T cells for 6d. The wild-type HL-60 cells were added and co-cultured for another 12h. Cytotoxicity assay was measured by LDH release. Results (1) The fragment of BCG HSP70 was consistent with the theoretical value. DNA sequencing showed that the recombinant vector of pDisplay-HSP70 was correctly constructed. (2) BCG HSP70 expressed onto the HL-60 cells sur-face. (3) Detection of the immunogenicity: ①The most significant T cell proliferation was observed in the group of HSP70-transfected HL-60 cells (P<0.05). There was no difference between the HL60-wt group and HL60-pDisplay group (P>0.05).②The contents of IFN-γof the HSP70-HL60 group was the highest.③The inhibiting activity of CTLs on HL-60 cells in the group of HSP70-transfected HL-60 cells was more significant than that of wide-type and pDisplay--transfected HL-60 cells. And with the increase of the E:T ratio, the inhibiting activity of CTLs in the HSP70-HL60 group was rising. Conclusions The recombinant eukaryotic expression vector (pDisplay-HSP70) of BCG HSP70 was successfully constructed. And the HL-60 cell vaccine expressing BCG HSP70 onto its surface was successfully prepared. The results showed that gene transfection of BCG HSP70 could significantly enhance the immunogenicity of HL-60 cells.

Prostate cancer is one of the life threatening disorders for human being, which has a high incidence over the world. At present, treatments include surgery, radiation therapy, cryosurgery, chemotherapy and hormonal therapy etc. Also, dietary modifications may decrease incidence of developing prostate cancer. This article summarizes current treat-ments, which may be helpful to prevent life threatening disease.

Objective To compare the transurenthral resection to radical cystectomy on muscle invasive bladder cancer, and to explore the factors affecting the prognosis. Methods Data of 74 patients with muscle invasive bladder can?cer were retrospectively analyzed. There were 38 cases underwent radical cystectomy (group A), and were treated with intra?venous chemotherapy after operation. There were 36 cases underwent transurenthral resection (group B), and were treated with intravenous and urinary bladder irrigation chemotherapy. All patients were followed up 61 (5-91) months. Data were compared between two groups including duration of surgery, intraoperative blood loss, the cumulative length of hospital stay, cancer recurrence rate and 5-year survival rate . The factors may affecting the prognosis in patients were collected and ana?lyzed by the Log-rank univariate and Cox multivariate analyzed. Results The values of operation time, intraoperative blood loss and the cumulative length of hospital stay were significantly lower in group B than those of group A ( P<0.01). There were no significant differences in cancer recurrence rate and 5-year survival rate between two groups (P>0.05). Results of the Log-rank univariate analysis showed that the tumor size≥5 cm and T3 stage were the important factors of 5-year relapse-free survival rate and 5-year overall survival rate. Results of the Cox multivariate analysis showed that the tumor size≥5 cm (RR=3.687, 95%CI:1.913-7.105, P<0.001) was the important factor of recurrence in patients after operation. T3 stage (RR=3.325, 95%CI:1.437-7.695, P=0.005) and tumor size≥5 cm (RR=5.017, 95%CI:2.440-10.317, P=0.002) were the risk fac?tors of the 5-year overall survival rate. Conclusion The transurenthral resection with intravenous and urinary bladder irri?gation chemotherapy deserves recommendation for the treatment of muscle invasive bladder cancer. Tumor size ≥5 cm and T3 stage are the important factors for the prognosis.

Objective To culture the acute leukemia cells in vitro,and to prepare cancer vaccine expressing heat shock protein 70 (HSP70) of Bacille Calemette-Geérin(BCG) onto the cell surface,so as to study its anti-tumor effect and mechanism.Methods Acute myeloid leukemia (AML) cells were cultured in a serum-free Stemspan(H) culture supplemented with cytokines [stem cell factor(SCF),flt-3 ligand (FL),interleukin (IL)-3 and IL-6] in vitro.And B-lineage acute lymphoblastic leukemia (B-ALL) cells were cultured in a Iscove modified medium(IMDM) culture supplemented with cytokines (SCF,FL,IL-3 and IL-7) in vitro.Cellular morphology was observed by the microscopy and immunophenotype determination was used to verify the biological characteristics of acute leukemia cells after culture.Lipofectamine 2000 was used to transfect the pDisplay-HSP70 plasmid into acute leukemia cells.The expression of HSP70 on the cell surface was detected by fluorescene microscope.Then the immunogenicity of the leukemia cells expressing HSP70 were detected.The experimental groups were divided into 3 subgroups:the wide-type acute leukemia cells (wt-LC group),the pDisplay-leukemia cells (pDisplay-LC group),and the pDisplay-HSP70-leukemia cells (HSP70-LC group),respectively.The leukemia cells in different groups were cultured with autologous peripheral blood T cells for 72 hours.The proliferation indices of T cells were assayed by carboxyfluorescein diacetate succinimidyl ester (CFSE)-staining method,and the contents of interferon-γ(IFN-γ) were tested by enzyme-linked immunosorbent assay (ELISA).The leukemia cells in different groups were cultured with autologous peripheral blood T cells,and after 6 days,the fresh acute leukemia cells were added [in the different ratios of cytotoxicity T lymphocyte (CTL):leukemia cells were 10 ∶ 1,20 ∶ 1,40 ∶ 1 and 80 ∶ 1] and continued to be cultured for another 12 hours.Cytotoxicity assay was measured by lactate dehydrogenase (LDH) release.Results After short term culture in vitro,the leukemia cells were in colony-like suspension and maintain the proliferation characteristics were maintained.The cell proliferation was rapidly cultured for about 10 days and then was gradually slowed down.But there was no difference between the day 10 and day 0 in the expressions of CD13 and CD33 in fifteen cases of AML cells (P ＞ 0.05).Equally,there was no difference between the day 10 and day 0 in the expressions of CD19,CD10 and CD22 in fifteen cases of B-ALL cells (P ＞ 0.05).After BCG HSP70 gene transfection,the yellow-green fluorescence on the leukemia cells surface was observed under the confocal microscope.Detection of the immunogenicity:(1) Autologous T cell proliferation:the most significant T cell proliferation was observed in the group of HSP70-transfected leukemia cells (t =17.89,19.58,all P ＜0.05).There was no difference between the wt-LC group and pDisplay-LC group (P ＞ 0.05).(2) The contents of cytokines:the IFN-γ level in the group of HSP70-transfected leukemia cells was higher than those of wide-type acute leukemia cells and the pDisplay-transfected ones (t =24.72,24.81,all P ＜ 0.05).(3) Cytotoicity of CTL:the killing rate in HSP70-transfected leukemia cells was significantly higher than those of wide-type acute leukemia cells and pDisplay——transfected ones(F =13.66,P ＜ 0.05).And with the increase of the ratio from 10 ∶ 1 to 80 ∶ 1,the inhibiting activity of CTL in the HSP70-LC group was raising(F =19.69,P ＜ 0.05).Conclusions Fresh acute leukemia cells can be successfully cultured in vitro.Short-term culture can significantly increase the number of leukemia cells,but has little effect on surface antigen expression.So,the biological characteristics of the leukemia cells can be maintained.The leukemia cells vaccine expressing BCG HSP70 onto its surface was successfully prepared,and gene transfection of BCG HSP70 can significantly enhance the immunogenicity of leukemia cells.

Objective The objective of this study was to investigate the expression of PSMD10 and Ki67 in epithelial ovarian tumors and its clinicopathological significance.Methods Immunohistochemical method was used to detect the expression of PSMD10 and Ki67 in 21 cases of ovarian benign tumor,31 cases of ovarian borderline tumor and 156 cases of ovarian cancer.Results The positive expression rate of PSMD10 and Ki67 in ovarian cancer was significantly higher than those of ovarian borderline and benign tumor(P<0.05).The positive expression rate of PSMD10 in ovarian cancer tissues was not different from others groups(P>0.05).There was significant different the positive expression rate of PSMD10 from clinical stage,pathological grade and residual tumor diameter(P<0.05).There was a positive correlation between the expression of PSMD10 and Ki67 in ovarian cancer(P<0.01).Conclusion PSMD10 and Ki67 play an important synergistic role in the carcinogenesis and progression of epithelial ovarian cancer.It can be used as an important index to judge the degree of malignancy and progression of ovarian cancer.Joint testing PSMD10 and Ki67 are used to guide the clinical diagnosis.

OBJECTIVE:To establish a method for the determining contents of 2 lignan components[dehydrodiisoeugenol and 2,3-dihydro-7-methoxy-2-(3,4-methylened ioxyphenyl)-3-methyl-5-(E)-propenyl-benzofuran(referred to“lignanoid 2”)]. METH-ODS:HPLC method was adopted. The column was Elite C18 with the mobile phase of water-methanol(gradient elution)at the flow rate of 1.0 ml/min;the detection wavelength was 225 nm and the column temperature was 30 ℃. The sample size was 20 μl. RE-SULTS:There was a good linear relationship between sample quantity and the peak area in the range of 0.202-2.02 μg(r=0.999 9) and 0.204-2.04 μg(r=0.999 9)for 2 lignan components. The RSD of precision,stability and repeatability tests were less than 2%with the average recovery of 101.54%(RSD=0.60%,n=6)and 99.43%(RSD=1.09%,n=6). CONCLUSIONS:The method is simple,sensitive and accurate,and can be used for the quantization determination of dehydrodiisoeugenol and lignanoid 2 in nut-meg-5.

Objective To assess the related risk factors of endoscopic retrograde cholangiopancreatography(ERCP)on postoperative pancreatitis.To improve the level of diagnostic and therapeutic ERCP,to reduce the incidence of postoperative pancreatitis.Methods A total of 346 patients in our hospital referred to diagnostic and therapeutic ERCP(399 frequency)were divided into 8 groups;the differences of postoperative serum amylase in 24 hour as well as clinical symptoms were compared among different groups.Results The incidence of postoperative hyperamylasaemia was 12.5 %.The incidence of postoperative acute pancreatitis was 1.5 %.ERCP+STENT therapeutic group(38.9%)had the highest incidence of postoperative hyperamylasaemia and postoperative acute pancreatitis among the groups.(respectively 38.9%,11.1%).There was the different incidence of postoperative actcte pancreatitis between the period from June 2003 to June 2005 and from July 2005 to July 2007,respectively 3.3%,0.4%.Conclusion Pancreatic duct contrast filling and deficient experience of doctor during ERCP manipulation are the main risk factors for postoperative pancreatitis.

Objective:To investigate the relationship of explicit self-esteem,implicit self-esteem,rumination and depression in a sample of adolescents.Methods:171 adolescents were investigated by Rosenberg Self-esteem Scale,CES-D,Ruminative Responses Scale and self-esteem IAT.Results:Pearson's correlation analysis showed a significant and negative relationship between explicit self-esteem and rumination (r=-0.394,P