ABSTRACT
With various diseases ravaging internationally, the demands for recombinant adenoviral vector (Adv) vaccines have increased dramatically. To meet the demand for Adv vaccine, development of a new cell culture process is an effective strategy. Applying hyperosmotic stress in cells before virus infection could increase the yield of Adv in batch culture mode. Emerging perfusion culture can significantly increase the yield of Adv as well. Therefore, combining the hyperosmotic stress process with perfusion culture is expected to improve the yield of Adv at high cell density. In this study, a shake flask combined with a semi-perfusion culture was used as a scaled-down model for bioreactor perfusion culture. Media with osmotic pressure ranging from 300 to 405 mOsm were used to study the effect of hyperosmotic stress on cell growth and Adv production. The results showed that using a perfusion culture process with a hyperosmotic pressure medium (370 mOsm) during the cell growth phase and an isosmotic pressure medium (300 mOsm) during the virus production phase effectively increased the yield of Adv. This might be due to the increased expression of HSP70 protein during the late phases of virus replication. The Adv titer in a bioreactor with such a process reached 3.2×1010 IFU/mL, three times higher than that of the traditional perfusion culture process. More importantly, this is the first time that a strategy of combining the hyperosmotic stress process with perfusion culture is applied to the production of Adv in HEK 293 cells. It also reveals the reason why the hyperosmotic stress process increased the yield of Adv, which may facilitate the process optimization of for producing other Adv in HEK 293 cells.
Subject(s)
Humans , HEK293 Cells , Genetic Vectors/genetics , Batch Cell Culture Techniques , Bioreactors , PerfusionABSTRACT
OBJECTIVE:To investigate the efficacy and safety of Xiaojin pill combined with docetaxel,epirubicin and cyclo-phosphamide in the treatment of breast cancer in stage Ⅲ and its effect on immune functions. METHODS:102 patients with breast cancer in stage Ⅲ were randomly divided into control group (51 cases) and observation group(51 cases). All patients were given symptomatic treatments as hydration,alkalization and antiemetic treatments in the duration of chemotherapy,and were given conven-tional treatments as 0.75 mg Dexamethasome oral tablets in 12 and 6 hours before the chemotherapy lespectively. Control group was given 75 mg/m2 Docetaxel injection,d1,intravenous infusion+60 mg/m2 Epirubicin hyclrochloride for injection,d1,intravenous in-fusion+600 mg/m2 Cyclophosphamide for injection,d1,intravenous infusion;observation group was additionally given 6 g Xiaojin pill,orally,twice a day. 21 d was regarded as a treatment course. Short-term clinical efficacy,immunoglobulin,T lymphocyte sub-sets(CD4+/CD3+,CD8+/CD3+,CD4+/CD8+)and the incidence of adverse reactions before and after chemotherapy in 2 groups after 2 courses were observed. RESULTS:There were no significant differences in the short-term effective rate and incidence of adverse reactions between 2 groups (P>0.05). Before chemotherapy,there were no significant differences in the immunoglobulin and T lymphocyte subsets between 2 groups(P>0.05). After chemotherapy,immunoglobulin levels in 2 groups were significantly lower than before,the differences were statistically significant (P0.05);CD4+/CD3+,CD8+/CD3+,CD4+/CD8+ in control group and CD8+/CD3+ in observation group were significantly lower than before,and CD4+/CD3+ and CD4+/CD8+ in observation group were higher than control group,the differences were statistically significant (P0.05). CONCLUSIONS:Based on the conventional treatment,Xiaojin pill combined with docetaxel,epirubicin and cyclophosphamide shows significant effi-cacy in the treatment of breast cancer in stageⅢ,and can improve immune functions,with good safety.
ABSTRACT
Objective To investigate the distribution characteristics of bone marrow(BM)cell morphological results and their role in the etiological diagnosis of blood diseases,and to understand the characteristics of hematological diseases spectrum in this ar-ea.Methods The BM puncture specimens in 372 cases of hematological diseases in our hospital from March 2012 to April 2014 were performed the Wright-Giemsa staining and cytochemical staining.The cell morphology and cytochemical staining were ob-served by microscope,which was combined with the related clinical data for obtaining the morphological report.Results Among 372 cases,368 cases were diagnosed with hematological diseases,which were dominated by 3 kinds of main type,leukemia(95/372), hyperplasia anemia(36/372)and iron-deficiency anemia (26/372).In 82 cases of anemia,hyperplasia anemia(36/82),iron-deficiency anemia (26/82)and megaloblastic anemia (11/82)were predominant.Conclusion The BM morphological examination is one of im-portant measures for the etiological diagnosis in hematological diseases.The analysis on the diseases spectrum is conducive to guide the diagnosis and treatment in clinic.