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.

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 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.

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

OBJECTIVEThis study aims to investigate the expression levels of serum and urinary vascular endothelial growth factor-A (VEGF-A) and epidermal growth factor-like domain 7 (EGFL7) in proliferating infantile hemangioma patients under propranolol treatment.

METHODSPropranolol (0.5-2 mg x kg(-1)) was orally administered to 30 infants every day for 4-8 months. The Achauer method was used to measure the tumor radius and thus evaluate the clinical curative effects of the treatment. Enzyme-linked immunosorbent assay was used to measure the serum and urinary concentrations of VEGF-A and EGFL7 at 0, 4, and 12 weeks after the treatment.

RESULTSThe treatment response was excellent in 2 patients, good in 11, moderate in 14, and poor in 3. Serum VEGF-A (335.692 pg x mL(-1) ± 136.146 pg x mL(-1)) was high before the treatment and then significantly decreased after 4 weeks (264.853 pg x mL(-1) ± 122.120 pg x mL(-1)) and 12 weeks (211.345 pg x mL(-1) ± 104.035 pg x mL(-1)) of treatment (P < 0.05). Urinary VEGF-A (76.234 pg x mL(-1) ± 24.169 pg x mL(-1)) was high before the treatment and then significantly decreased after four weeks (56.454 pg x mL(-1) ± l6.111 pg x mL(-1)) and twelve weeks (34.728 pg x mL(-1)) ± 12.656 pg x mL(-1)) of treatment (P < 0.05). Serum and urinary EGFL7 also decreased after the treatment, showing a positive relationship with VEGF-A.

CONCLUSIONPropranolol can be safely and effectively used to treat proliferating infantile hemangiomas. This treatment can reduce the peripheral serum and urinary concentrations of VEGF-A and EGFL7 in affected children.

Objective To evaluate the clinical efficacy of propranolol in treating proliferating infantile haemangiomas,and to measure the expression levels of vascular endothelial growth factor-A (VEGF-A) and hypoxiainducible factor 1α (HIF-1α) in sera and urine of patients during the treatment.Methods Thirty infants with proliferating haemangiomas were treated with propranolol at doses of 0.5-2 mg/kg per day.The radius of haemangiomas was measured,and blood and urine samples were obtained from these patients before,and at 4 and 12 weeks after the beginning of treatment.Clinical efficacy was estimated according to a four-graded scale as well as the feedback from parents of these patients.Enzyme-linked immunosorbent assay (ELISA) was performed to determine the serum and urine concentrations of VEGF-A and HIF-1α.Thirty check-up infants collected from the Department of Child Health Care served as the healthy controls.Statistical analysis was done by two-way analysis of variance followed by the least significant difference (LSD) test.Results After 12 weeks of treatment,clinical response was excellent in 2 patients,good in 11,moderate in 14,and poor in 3.The serum levels of VEGF-A and HIF-1α were (268.174 ± 95.056) μg/L and (10.809 ± 1.686) mg/L respectively in the control group,sequentially decreased in the patients from baseline to 4 and 12 weeks after the beginning of treatment (VEGF-A:(385.692 ± 136.146) vs.(264.853 ± 122.12) vs.(211.345 ± 104.035) μg/L; HIF-1α:(31.462 ± 7.458) vs.(21.454 ± 5.489) vs.(12.052 ± 3.623) mg/L).The trend in expression changes of VEGF-A and HIF-1α in urine samples was similar to that in blood samples in these patients.Positive correlation was observed between the expression level of VEGF-A and HIF-1α in sera (r=0.730,P＜ 0.05) and urine (r=0.667,P＜ 0.05) of these patients.Moreover,the levels of serum VEGF-A,urine VEGF-A,serum HIF-1α and urine HIF-1α were all negatively correlated with the time course following propranolol administration (r =-0.390,-0.689,-0.806,-0.683,P ＜ 0.05,0.01,0.05,0.01 respectively).Conclusion Propranolol is effective for the treatment of proliferating infantile haemangiomas,likely by reducing serum and urinary concentrations of VEGF-A and HIF-lα in children.

Two isolation methods for sorting of endothelial progenitor cells (EPCs): from peripheral blood mononuclear cells (PBMCs) and CD133(+) enriched cells were compared, by defining the cell morphology, phenotype, reproductive activities and function in vitro, to provide a reference for clinical application of EPCs. PBMCs from healthy subjects were used either directly for cell culture or for CD133(+) sorting. The two groups of cells were cultured in complete medium 199 (M199) for 7 to 14 days and the phenotypes of EPCs were analyzed by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and the VEGF concentration was measured using an ELISA kit. ECM gel experiment and migration assay were performed in vivo. The results showed that PBMCs produced more colony-forming units (CFU) than CD133(+) enriched cells from the same volume of blood (P<0.01). From day 7 to 14, the two groups showed decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144(+) cells in CD133(+) group were less than in PBMCs group (P<0.01). PBMCs group secreted more VEGF than CD133(+) group on the day 7 (P<0.01). As compared with CD133(+) group, PBMCs group had more potent potential of proliferation and vascularization in vitro. It was concluded that CD133(+) sorted cells showed a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, suggesting that CD133-negative cells may be a preferential way to get EPCs for clinical therapy.

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.

BACKGROUND: Documents recorded that the correlation between micro emulsion Ciclosporin peak concentration (C_2) and area under curve was best with maximum individual difference. According to C_2, dose of Ciclosporin can be adjusted indMdually to decrease acute rejection and Ciclosporin toxicity, which has widely used in perioperative stage of renal transplanted recipients. However, some transplantation center still used tough concentration (C_0) to adjust the dose of Ciclosporin in stable stage of renal transplanted recipients. OBJECTIVE: To analyze the efficacy and safety of changing from monitoring C_0 to C_2 in stable stage recipients following renal transplantation. METHODS: Totally 65 patients with renal transplantation were enrolled in this study, including 31 males and 34 females, aged 20-57 (39.4±15.3) years. Within 3 months prior to this study, all patients did not suffered from rejection, and their serum creatinine and urea nitrogen were stable (creatinine ≤180 μmol/L). They were in stable stage after renal transplantation. Their period of transplantation and function of allograft were recorded. Their C_0 and C_2 of Ciclosporin were assayed. According to the target C_2 value 500-600 μg/L, the patients were prospectively and randomly divided into 3 groups. In the high C_2 group (n=17), the dose of Ciclosporin was decreased. In the target C_2 group (n=23), the dose of Ciclosporin was remained. In the low C_2 group (n=25), the dose of Ciclosporin was increased. All of the patients were followed-up for 12 months. The grafts function and the complications of heart, lung and brain were compared. RESULTS AND CONCLUSION: According to the target concentration of Ciclosporin C_2, the dose of Ciclosporin in the high C_2 group was decreased by 575.0 mg. The Creatinine and urea nitrogen of 88% patients were stable, while blood pressure, blood fat and blood uric acid decreased in parts of patients. In the target C_2 group, the levels of creatinine, urea nitrogen, Co and C_2 of patients were stable, no complications of heart, lung and brain occurred. According to the target concentration of Ciclosporin C_2, the dose of Ciclosporin in low C_2 group was increased by 755.0 mg. The creatinine and urea nitrogen of 84% patients were stable. All of the patients were no complications of heart, lung and brain. It is safe and effective to adjust Ciclospori dose under C_2 monitoring according to the target peak concentration (500-600 μg/L) in most stable stage recipients following renal transplantation.