Objective To address the standard first-line management under the new diagnostic criteria in adult immune thrombocytopenia (ITP). Methods A retrospective analysis was conducted involving 178 adult ITP patients treated with high-dose dexamethasone or prednisone in Qilu Hospital from March 2004 to November 2009 using new diagnostic criteria. Results The median age was 41 years with a male/female ratio of 0. 73: 1. Among the 178 ITP patients, 87 were newly diagnosed, 30 persistent ITP, 58 chronic ITP, and 3 unable to follow up. The efficacy rates among 167 patients able to assess in the three groups were 77.4% ( 65/84 ), 64. 0% ( 16/25 ) and 62. 1% ( 36/58 ) respectively, and their complete remission (CR) rates were 57. 1% (48/84), 36. 0% (9/25) and 32. 8% (19/58). The efficacy rate and CR rate of the newly diagnosed ITP category were significantly higher than those of the chronic ITP category (x2 = 3. 917, P ＜ 0. 05 ;x2 = 8. 186, P ＜ 0. 01 ). The patients treated with high-dose dexamethasone or prednisone therapy had no significant differences in sex, age or blood platelet count before treatment.Moreover, the short or long term response rates and the CR rates between the two therapies had no statistically significant differences while the former had a shorter onset time ( F = 10. 34, P ＜ 0. 01 ).Conclusions The study sets up a basis for the application of the recommended new definition and outcome criteria for adult ITP. Dexamethasone therapy is favored as first-line therapy.

Purpose: To investigate the effect of HSV-TK/GCV system on human prostate cancer. Methods: MTTassay, flow cytometry( FCM), optical and electron microscopy were used to determine the sensitivity of GCV on prostate cancer cell( PC-3m) after being tranfected with HSV-TK gene. Results: It was found that GCV had significant cytotoxic activity on HSV-TK gene-transduced prostate cancer cells, but little effect on the non-transduced cells. Conclusions: The experiment indicates that HSV-TK/GCV system has significant antitumor activities on the TK gene-transduced prostate cancer cells in vitro.

Objective To explore the effect of sodium fluoride(NaF) on the tumor markers of human embryonic bronchial epithelium (HEBE) cells.Methods HEBE cells were collected from the human abortive fetues.The cells were exposed to NaF of several concentrations for 36 h.After rinsed,the cells were incubated for 36 h again.The NaF toxicity to HEBE cells was detected using MTT method.The supernatant was collected and the lung tumor markers were detected using ELISA method.Results NaF was toxic to the HEBE cells,and the toxicity was increased with the NaF doses.There were few survival HEBE cells at 6 mmol/L NaF.The tumor markers in both of the control and experiment groups were very low,and no significant difference had been seen between them.Conclusion NaF may damage HEBE cells,but may not influence the tumor markers of HEBE cells.

Objective To investigate the role of Th17 cells in chronic myeloid leukemia (CML) pathogenesis. Methods 33 CML patients [15 newly diagnosed (ND)- and 18 chronic-phase (CP)- CML patients] and 15 healthy controls were enrolled. The percentage of Th17 cells in the peripheral blood (PB) of CML and controls were evaluated by flow cytometry. RORC mRNA expressions were examined by real-time quantitative reverse transcription-polymerase chain reaction (RT-PCR). The levels of IL-17 in PB of CML and controls were measured by enzyme-linked immunoadsorbent assay (ELISA). The relationships between Th17 cells and clinical characteristics in CML were analyzed. Results The percentage of Th17 cells in PB of ND-CML patients [(0.71±0.41) %] was significantly lower than that of CP-CML patients [(4.08±0.74) %, P<0.05] and healthy controls [(3.18±1.32) %, P<0.05]. The mRNA level of RORC in PB of ND-CML patients (0.043 3±0.040 5) was significantly decreased compared with that in CP-CML patients (0.086 1±0.052 3, P<0.05) and healthy controls (0.091 0±0.058 4, P<0.05). The levels of IL-17 in PB of ND-CML patients [(1.43±0.22) pg/ml] and CP-CML patients [(1.36±0.19) pg/ml] were slightly higher than that of healthy controls [(1.23±0.14) pg/ml, P< 0.05]. The percentage of Th17 cells had significantly negative correlations with white blood cell counts in PB or bcr-abl (IS). Conclusion Th17 cells may play an important role in CML pathogenesis, which has potential implication for immunotherapy of this malignancy.

Objective To investigate the repressing effects of cytosine deaminase(CD) and herpes simplex(virus) thymidine kinase(HSV-tk) double suicide genes coexpression system on human cholangiocarcinoma QBC939 cells and QBC939 cells transplantated tumor in nude mice.Methods CD and HSV-tk double suicide genes were transfered into QBC939 cells using liposomes.After G418 selection,the positive clones of QBC939/CD+tk cells were picked up and cultured.The expression of CD and HSV-tk genes was(confirmed) by RT-PCR.In vitro,the QBC939/CD+tk cells were treated with 5-Fc and/or GCV,and the cytoxicity efficacy was evaluated by microculture tetrajolium test(MTT) method.The QBC939/CD+tk cells were inoculated subcutaneously into nude mice,and when the tumors were palpable,5-Fc and GCV were(injected) intraperitoneally,and the volumes of transplantated tumors were measured before and after medication.Results Double suicide genes were stably expressed in QBC939/CD+tk cells.The repressing capability of combination of 5-Fc and GCV on QBC939/CD+tk cells was more effective than that of using either 5-Fc or GCV alone.The increase of cell-repressing was assaciated with increase the concentration of the prodrug.The repressing effect of combination of the 2 prodrugs on early period of transplantation tumor was obvious,even complete abolition of tumor was noted,and moreover a marked local bystander effect was observed.(Conclusions) In vitro and in vivo,the cell-repressing efficacy of double suicide gene system on cholongiocarcinoma cells and the tramsplanted tumor of nude mice was significant,and the bystander effect was obvious.

Objective To study the in vitro effects of sonodynamic management on human ovarian cancer cell line HRA. Methods Hematoporphyrin was selected as a sonosensitizer and ultrasound with certain intensity was used to activate hematoporphyrin.Then MTT assay and clony-forming efficiency assay were applied to determine the growth inhibitory effects of ovarian cancer cells. Electron microscope was applied to detect the morphological changes of the cells. Cell apoptosis was analyzed using flow cytometry. Results Hematoporphyrin had no significant inhibitory effects on the cells. However, when hematoporphyrin was used before ultrasound,it could enhance the killing effects of ultrasound(P

Objective:To assess the potential of mouse immune-costimulating signal B7-2 in inducing immune effect.Methods:(1)The specific mB7-2cDNA fragment from LPS-stimulated mouse splenocytes was obtained by reverse transcription and polymerase chain reaction.The obtained fragment was inserted to pLXSN plasmid.Then the plasmid pLXSN-mB7-2 was packaged into PA317 cells;(2)The EL4/mB7-2 was obtained by infecting the EL4 by the concentrated virus particles produced by PA317/mB7-2;(3)In vitro,the secretion of IL-2 of EL4/mB7-2 stimulated-lymphocytes was detected by using mixed lymphocytes culture.Results:pLXSN-mB7-2 and transgenetic EL4/mB7-2 cells were obained successfully.IL-2 production in 24h in the supernatant stimulated by EL4/mB7-2 was much higher than wild EL4 cells.Conclusion:EL4/mB7-2 can activated T cell to produce IL-2.This research lay foundation for the research of the function of immune-costimulating signal in the tumor immunity and treatment,the mechanism of autoimmune disease and organ transplantation.

Objective: To investigate effect of exogenous wild-type p53 gene on the cell growth and tumorigenicity of human gallbladder cancer cell lines. Methods: After identification of the genetic status of p53 gene of GBC-SD cell lines with the immunocytochemistry staining and the direct sequencing technique of PCR products, eukaryotic expressing plasmid pCMV-p53 was introduced by lipofectamine-mediated into GBC-SD cell lines. Growing transfected cells were selected by G418. The presence and expression of exogenous p53 gene was detected by PCR, RT-PCR and Western blot. The cellular proliferating ability was assessed using the cell growth curve and cloning assay. The xenograft in nude mice was performed to examine the effect of tumorigenicity. Results: P53 protein overexpression was showed in GBC-SD cell lines. A transversion of TAC→AAC at codon 126 of exon 5 was confirmed. PCR, RT-PCR and Western blot showed exogenous p53 gene had successfully transfected into GBC-SD cells and obtained high expression. The growth and proliferation of the cells were greatly decreased, and the tumorigenicity was significantly inhibited after transfection wtp53. Conclusion: The expression of exogenous wild-type p53 gene could effectively inhibit the growth of gallbladder cancer GBC-SD cells in vitro and in vivo.

Objective:To investigate the different killing effects on human hilar cholangiocarinoma cells FRH with cytosine deaminase(CD) and herpes simplex virus thymidine kinase(HSV tk)double suicide genes coexpression compared with single gene mediated by retrovirus.To find a more efficient and low toxicity suicide gene therapy for hilar cholangiocarinoma.Methods:CD and HSV tk double suicide genes were transfected into PA317 cells using lipofectamine.The positive clones were picked out and cultured after G418 selected.The viral supernatant was collected.The FRH cells were infected with the virus containing the double suicide genes.After G418 selection,RT PCR was resorted to demonstrated the successful transcription of CD and HSV tk genes.The FRH/CD+tk and FRH cells in culture were respectively treated with 5 Fc and /or GCV.The cytoxicity efficacy was evaluated by microculture tetrajolium test (MTT) method.Results:The virus containing double suicide gene was produced in PA317 cells.Double suicide genes were stably expressed in RFH cells after being infected with the virus.The killing effect of combination 5 Fc with GCV on FRH/CD+tk cells is more effective than that of using 5 Fc or GCV alone.Conclusion:The CD+TK/5 Fc+GCV co expression system is more effective for killing effects on FRH cells than that of CD/5 Fc or tk/GCV system alone.

Objective To construct an eukaryotic expression vector of retinoblastoma gene (pIRES-Rb94), and investigate the radiosensitising effect of Rb94 on lung adenocarcinoma cell line A549 and the mechanism. Methods Recombinant expression plasmid pIRES-Rb94 was constructed and then transfected into A549 cells using lipofectamine 2000. Steadily transfected cells were obtained using G418 se-lecting system. Cell counting method was used to produce the growth curve and the population doubling time was then calculated. The radiosensitivity of A549 cells was assessed by clonogenic assay. The expression of bTERT and Bcl-2 mRNA was detected by real-time RT-PCR. Cell cycle distribution was measured by flow cytometry. Results Steadily transfected cell line pIRES-Rb94-A549 was aquired. Compared with A549 cells, the population doubling time of pIRES-Rb94-A549 cells was increased from 31.5 h to 39.5 h (t=5.15, P<0.01). The expression of hTERT and Bcl-2 mRNA was both down-regulated (0.02:1.00, t= 18.99,P<0.01,0.01:1.00,t=13.73,P<0.01). The number of cells was increased in G2/M phases (35.91%:4.53%, t =36.78,P=0.00), whereas decreased in G0/G1 and S phases (47.02%:74.07%, t =11.71,P=0.00;17.07%:22.32%, t =2.30,P<0.05). The sensitizing enhancement ratio was 1.30. Conclusions Rb94 has marked radiosensitizing effect on A549 cells by G2/M phase blockage and down-regulation of hTERT and Bcl-2 mRNA expression. Rb94 may also inhibit the ability of cell proliferation by regulating cell cycle distribution.