Objective To optimize the conditions for enhancing the refolding of a novel recombinant human(rh)endostatin and test the biological activities of refolded endostatin.Methods The partial purified inclusion bodies of rh-endostatin were dissolved with 6 mol/L guanidine-HCl followed by combination of dilution and dialysis of the dissolved endostatin.The refolded endostatin was then purified by cation-exchange chromatography.The biological activities of purified rh-endostatin were assessed by endostatin-specific monoclonal antibody and chick embryo chorioallantoic membrane assay.Results A 46% refolding yield was achieved after optimizing the refolding conditions.The purified endostatin reacted with specific anti-endostatin monoclonal antibody and showed significant inhibition of angiogenesis in chick embryo ehorioallantoic membrane assay.Conclusion The method of highest refolding yield of human endostatin was developed.This optimized method significantly promotes the application of this novel human endostatin to preclinical and clinical studies.

This review focus on the perspective of genetic engineering technology in biopharmaceutical field.Four general aspects of genetic engineering are further discussed in this review as following: the separation of macromolecules,PCR,gene chip,and the expression of foreign genes.The large-scale or industrialization of genetic engineering technology in biopharmaceutics is also reviewed.

Objective To explore the change of proportion of peripheral blood dendritic cells (DCs) in patients with stroke. Methods 56 patients (30 cases of cerebral infarction and 26 cases of cerebral hemorrhage) in Beijing Bo'ai hospital from June to September, 2014 and 14 healthy controls were investigated. The severity of stroke was assessed with the National Institutes of Health Stroke Scale (NIHSS). Flow cy-tometry analysis was employed to detect the proportion of DCs subtypes in the peripheral blood. Results No obvious difference was found in DCs between the stroke patients and the controls. Compared to the control group, the percentages of peripheral blood myeloid dendritic cells (mDCs) decreased in the cerebral hemorrhage and the cerebral infarction subgroups (P<0.001). The percentages of plasmacytoid den-dritic cells (pDCs) reduced significantly in the cerebral hemorrhage and the cerebral infarction subgroups (P<0.05). The stroke patients were divided into NIHSS≤7 subgroup and NIHSS>7 subgroup. The percentages of pDCs in the cerebral hemorrhage and the cerebral infarction patients were significantly lower in the NIHSS>7 subgroup than in the NIHSS≤7 subgroup (P<0.05). While there was no statistical differ-ence between NIHSS≤7 subgroups and NIHSS>7 subgroups in the percentages of mDCs in the cerebral hemorrhage and cerebral infarction patients. Conclusion The proportion of DCs subtypes in the peripheral blood in stroke patients changed significantly, indicating inflamma-tion responds play a role in stroke.

Objective To optimize the HT22 cell oxygen-glucose deprivation/re-oxygenation models in hippocampal neurons of mice,discuss the basis of model establishment and test the related parameters.Methods HT22 cell growth curve was plotted by cell counting and cell doubling time was calculated,and then,the oxygen-glucose deprivation time and re-oxygenation time were determined by LDH assay.The CCK-8 assay,cell counting,cell morphology and flow cytometry were performed to further optimize the parameters of the models.Results The HT22 cell doubling time was 17 h 18 min.The oxygen-glucose deprivation time was 18 h and the re-oxygenation was 20 h.CCK-8 assay indicated that absorbance in cells of oxygen-glucose deprivation 18 h/re-oxygenation 20 h (OGD 18 h/R20 h) group was 0.334±0.138,which was significantly lower than that in the control group (0.756±0.026,P＜0.05);cells in the OGD18 b./R20 h group were shrank obviously.Flow cytometry indicated that the percentage of cells in terminal apoptosis or non-viable non-apoptotic cells was 24.56％±5.76％ and that of viable apoptotic cells was 2.63％±0.55％ in the OGD18 h/R20 h group,which were significantly higher than those in the control group,respectively (8.13％±0.87％ and 0.80％±0.10％,/P＜0.05).Conclusion The HT22 cell oxygen-glucose deprivation/re-oxygenation models are successfully established;both oxygen-glucose deprivation time and re-oxygenation time are related to the cell doubling time,and OGD18 h/R20 h can preferably help the establishment;.

Mitochondrial shape rapidly changes by dynamic balance of fusion and fission to adjust to constantly changing energy demands of cancer cells. Mitochondrial dynamics balance is exactly regulated by molecular motor consisted of myosin and actin cytoskeleton proteins. Thus, targeting myosin-actin molecular motor is considered as a promising strategy for anti-cancer. In this study, we performed a proof-of-concept study with a natural-derived small-molecule J13 to test the feasibility of anti-cancer therapeutics