BACKGROUND:The form and structure of antigen-extracted xenogeneic cancel ous bone through series of physical and chemical treatment are similar to human tissue. OBJECTIVE:To detect the biocompatibility of antigen-extracted xenogeneic cancel ous bone matrix prepared by three different ways. METHODS:The antigen-extracted xenogeneic cancel ous bone scaffold materials which were prepared through physical, chemical and physical-chemical combined methods and hydroxy apatite biological ceramic materials were implanted into the dorsum subcutaneous tissue. Histological observation was done at 4, 8 and 12 weeks after surgery. The antigen-extracted xenogeneic cancel ous bone scaffold materials which were prepared through physical, chemical and physical-chemical combined methods respectively was used to culture sheep bone marrow mesenchymal stem cells for 7 days. Cel adhesion, growth, proliferation and stroma secretion were observed. RESULTS AND CONCLUSION:At 4 weeks after surgery, a strong inflammatory reaction was detected around materials in four groups. At 12 weeks, the xenogeneic bone materials prepared through physical and physical-chemical combined methods and hydroxy apatite biological ceramic materials internal pore and surrounding tissue inflammation disappeared basical y, with the presence of thimbleful inflammation cells. The material degradation was more than at 8 weeks. The xenogeneic bone materials prepared through chemical methods material internal pore and surrounding tissue inflammation stil existed, suggesting that the xenogeneic bone materials prepared through physical and physical-chemical combined methods exhibited good histocompatibility. A smal amount of orderly osteoblasts existed around hydroxy apatite biological ceramic materials and physical-chemical prepared materials, with a smal amount of bone. These suggested that there was a tendency for ectopic bone formation. The xenogeneic cancel ous bone materials prepared through physical or physical-chemical combined methods have better cytocompatibility. However, scaffold materials prepared through chemical method have poor cytocompatibility and they are not qualified for the safety standards of biological materials.

Survivin is the most recent member of the IAP family, which shows a higher expression in almost all tumor cells, but not detected in most normal cells. Our attention is drawn by survivin due to its selective expression, which was taken as one molecule sign of tumor cells. Accordingly, survivin is examined in most leukemia cell line and leukemia specimen, whose expression also showed the association with leukemia. As in other tumor cells, survivin also plays an important role in the leukemogenesis. Meanwhile the detection of survivin can provide an useful prognosis information. This article is a brief overview of the development of survivin and its relationship with leukemia.

Objective Role of melanocortin receptor 4 (MCAR) in excitatory amino acid release from rat astrocytes in spinal cord. Methods Astrocytes were isolated from the spinal cord of newborn pathogen-free Wistar rats ( 1-3 days after birth) and cultured in serum-free Neurobasal/B27 liquid culture medium. After 4 passages the primary cultured astrocytes were randomly divided into 3 groups (6 wells each): group Ⅰ control (group C); group Ⅱ the astrocytes were exposed to TNF-α 10 μg/L (group T) and group Ⅲ the astrocytes were exposed to TNF-α 10 μg/L and HS014 (selective MC4R antagonist) 1 μmol/L (group TH). The astrocytes were incubated at 37 ℃ for 3 h. The supernatant was collected for determination of glutamic acid (Glu) and aspartic acid (Asp)concentrations by HPLC-MS/MS. Results TNF-α significantly increased Glu and Asp release from astrocytes in group T as compared with group C. The Glu and Asp concentrations were significantly lower in group TH than in group T. Conclusion MG4R is involved in the excitatory amino acid release from astrocytes in the spinal cord.

To explore and discuss the application of case teaching method for pharmaceutical administration science according to the actual teaching situation and the teaching experience of the authors. The teaching effects can be improved by the method, which is worthy of promotion and popularization.

Objective To investigate the effect of cyclosporine on matrix metalloproteinase-3 (MMP-3)’ s genetic expression on bladder cancer in rats induced with BBN and its clinical significance.Methods Twenty SD rats were divide into experimental group or control group randomly.Ten samples of SD rats bladder cancer induced with BBN and cyclosporine simultaneously and 10 samples of SD rats bladder cancer induced with BBN only as control were used to observe the effect of cyclosporine on MMP-3’ s genetic expression.Real time RT-PCR and immunohistochemistry stain were used to analyze MMP-3 mRNA and protein levels of bladder cancer in rats respectively.Results The MMP-3 mRNA median expression were 7.6 (4.2-9.1) in experimental group and 4.7 (2.8-7.7) in control group.The MMP-3 protein expression were 1 case with (-),4 cases (+),5 cases (++) in experimental group and 3 cases (-),4 cases (+),3 cases (++) in control group.The differences of MMP-3 mRNA and protein levels of bladder cancer between experimental group and control group were both significant (P ＜ 0.05).Conclusion Cyclosporine may stimulate the growth and development of bladder cancer through changing expression of some genes like MMP-3,and MMP-3 maybe become one of the targets of chemoprevention for post-transplantation bladder cancer.

Objective To investigate the effect of cyclosporine on signal transducer and activator of transcription 1 (STAT1) genetic expression on bladder cancer in rats induced by BBN and its clinical significance.Methods Twenty SD rats were divide into experimental group or control group randomly.Ten samples of SD rats bladder cancer induced with BBN and cyclosporine simultaneously as experimental group,and 10 samples of SD rats bladder cancer induced with BBN only as control.Real time RT-PCR and immunohistochemistry stain were used to detect STAT1 mRNA and protein level expressions of bladder cancer in rats respectively.Results The STAT1 mRNA median expression fold was 4.5 (2.1-6.6) in experimental group and 5.6 (3.4-8.5) in control group.The STAT1 protein expression were 5 cases with (-),3 cases (+),2 cases (++) in experimental group and 0 case (-),5 cascs (+),5 cases (++) in control group.The expression of STAT1 mRNA and protein level of bladder cancer between experimental group and control group were both significant different (P ＜ 0.05).Conclusions Cyclosporine may stimulate the growth and development of bladder cancer through changing expression of some genes like STATI,and STAT1 maybe become one of the targets of chemoprevention for post-transplantation bladder cancer.

ObjectiveTo investigate DNA loads and risk factors of BK virus infection in renal transplant recipients.MethodsWe developed a real-time PCR assay to quantitate BK virus loads in 80 patients receiving renal transplantation in our center,and correlation between the BK virus load and clinical course was analyzed.BK virus loads were measured in urine and plasma. Epidemiological features and risk factors of BK virus infection were analyzed.ResultsThe positive rate of BKV viruria and viremia in 80 renal recipients was 37.5％ (30/80) and 8.75％ (7/80),respectively.BKV loads were higher in renal allograft recipients whose age was more than 50 years old.BKV loads were observed in urine and plasma (compared with group whose age was less than 50 years,P=0.017 and 0.05,respectively).BKV DNA copies were higher in group Tac than that in group CSA (P＜0.05),and the peak of BKV load in serum appeared at14th and10th month after transplantation,respectively,but the peak in urine was ahead of that in serum,appeared at 2nd and 8th month,respectively.ConclusionSerial measurement of BKV viral loads by quantitative PCR is a useful tool in monitoring the course of BK virus infection.The ages of recipients (＞50 years) and using Tac + MPA can reactivate BK virus and then result in BKVAN in renal transplant recipients. Intensive BKV monitoring is necessary for these recipients.

Recombinant Fl-V (rFl-V) fusion protein is the main ingredient of the current candidate vaccine against Yersinia pestis infection, which has been under investigation in clinical trial in USA. We investigated the soluble expression conditions of rF1-V in Escherichia coli BL21 (DE3) that we constructed before. After scale-up and optimization of fermentation processes, we got the optimized fermentation process parameters: the culture was induced at the middle exponential phase with 50 µmol/L of IPTG at 25 °C for 5 h. Soluble rFl-V protein was isolated to 99% purity by ammonium sulfate precipitation, ion exchange chromatography, hydrophobic chromatography and gel filter chromatography. The protein recovery was above 20%. Protein identity and primary structure were verified by mass spectrometry and Edman sequencing. Results of purity, quality and western blotting analysis indicated that the target protein is a consistent and properly folded product. Furthermore, the immunogenicity of various antigens formulated with aluminum hydroxide adjuvant was evaluated in mice. Serum antibody titers of 4 groups including 20 µg rFl, rV and rFl-V and 10 µg rFl+10 µg rV, were assayed by ELISA after 2 doses. The antibody titers of anti-Fl with 20 µg rFl-V were obviously higher than titers with other groups; meanwhile there were no significant difference of anti-V antibody titers among them. These findings confirm that rFl-V would be the active pharmaceutical ingredient of the plague subunit vaccine.
Adjuvants, Immunologic ; Animals ; Antibodies, Bacterial ; blood ; Antibody Formation ; Antigens, Bacterial ; immunology ; Blotting, Western ; Chromatography, Ion Exchange ; Enzyme-Linked Immunosorbent Assay ; Mice ; Plague ; prevention & control ; Plague Vaccine ; immunology ; Recombinant Fusion Proteins ; immunology ; Vaccines, Subunit ; immunology ; Yersinia pestis

Objective To investigate the effects of cyclosporine on cell proliferation, apoptosis and angiogenesis of bladder cancer in rats induced with BBN.Methods Ten samples of SD rats bladder cancer induced with BBN and cyclosporine simultaneously and 10 samples of SD rats bladder cancer induced with BBN only as control were used to observe the effects of cyclosporine on cell proliferation, apoptosis and angiogenesis of bladder cancer in rats.Immunohistochemistry stain with PCNA, TUNEL and immunohistochemistry stain with CD31 were used to analyze cell proliferation,apoptosis and angiogenesis of bladder cancer in rats respectively. Results The differences of proliferation index (35.3±8.6)% and (28. 7±8.0)% respectively (P＜0.05), apoptotic index (2. 5±0.8)% and (4.3±1.3)% respectively (P＜0.05) and microvessel density 32.5±8.2 and 26.3±8.1 respectively (P＜0.05) between experimental group and control group were all significant.Conclusions Cyclosporine may stimulate the growth and development of bladder cancer through mechanisms of cell proliferation, apoptosis and angiogenesis.

Objective To investigate the signal transducer and activator of transcription 1(STAT1) and matrix metalloproteinase 3(MMP3)′s genetic expressions and their clinical significance on urothelial carcinoma after renal transplantation. Methods Fifty-one patients with urothelial carcinoma were recruited in this study. Sixteen of them who had renal transplant were in the experimental group and 35 of them without renal transplant were in the control group. All the cases had been proved postoperatively having transitional cell carcinoma by histopathological study. The human genome oligo arrays were used to analyze the gene expression spectrum of urothelial carcinoma after transplantation, aiming the STAT1 and MMP3's expression. Real time RT-PCR and immunohistochemical staining were used to compare the differences in the 2 groups. Results The experimental group showed that there were 35 genes up-regulated compared with the control group. Of them, 23had known gene function or partly known, and 12 had unknown gene function. There were 76 genes down-regulated. Of them, 46 had known gene function or partly known, and 30 had unknown gene function. After pathway analysis of the differentially expressed genes, there were 23 groups of pathways which had significant differences (P＜0.05), referring to the aspects of immunosuppressive and tumor growth. The levels of STAT1 and MMP3 expressions had significant differences between the 2groups(P＜0.05)as well. Conclusions The differential expression of urothelial tumor genes is obvious between patient who has had renal transplant and who has not. There are many aspects that are related to the tumor's growth like signaling pathways regulating proliferation, apoptosis of tumor cells, tumor angiogenesis and the tumor metastasis potential. STAT1 and MMP3 maybe become the targets of chemoprevention for post-transplantation urothelial carcinoma.