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Objective:To investigate the optimal dose of Th1/Th2 immuno-enhancement effects of cultivated Artemisia rupestris L. crude polysaccharides (CARCP) on foot-and-mouth disease vaccine (FMDV) via the intramuscular route. Methods:ICR mice were intramuscularly immunized twice with different concentrations of CARCP mixed with FMDV at 2-week intervals. FMDV-specific antibodies, isotypes, and IgE in serum were detected by ELISA. Splenocyte proliferation was detected by MTT. T lymphocyte subsets and cytokines in the spleen were detected by flow cytometry. Clinical signs and local reactions at the injection site were monitored daily, and the body weight of mice was weighed after immunization.Results:The medium dose of CARCP could significantly improve FMDV-specific IgG, IgG 1, and IgG 2a antibody levels and the IgG 2a/IgG 1 ratio ( P<0.05) and lead to significant splenocyte proliferative responses ( P<0.01). The medium dose of CARCP could also significantly increase the level of CD3 +CD4 + and CD3 +CD8 + T cells as well as CD4 +/CD8 + ratio ( P<0.05), elicited the higher levels of IFN-γ in CD4 + T cells and CD8 + T cells ( P<0.05). No local adverse reactions at the injection site were observed after immunization. There was no significant difference in body weight or growth between each group( P>0.05). CARCP did not significantly induce an IgE response ( P>0.05). Conclusions:CARCP as an FMDV adjuvant promotes Th1/Th2 immune responses, especially in favor of the Th1 response, and has a certain safety profile. The best immune enhancement is achieved with CARCP at medium doses.
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Objective To compare the immunopotentiating effects of polysaccharides extracted from wild/cultivate Cistanehe deserticola (WCDPS/CCDPS) in Xinjiang. Methods ICR mice were subcu-taneously injected twice with different doses(low,medium and high) of WCDPS and CCDPS in combination with ovalbumin (OVA). OVA-specific antibody IgG,as well as IgG1 and IgG2a subtypes, was determined by ELISA. OVA-specific lymphocyte proliferation was measured by MTT. Expression of CD4+T and CD8+T cells was analyzed by flow cytometry. Results Both WCDPS and CCDPS could significantly improve the production of OVA-specific IgG,IgG1 and IgG2a,promote the proliferation of OVA-specific lymphocytes and increase the expression of CD4+T and CD8+T cells(all P<0.05) with no significant difference between them at the same dosages (P>0.05). WCDPS and CCDPS had no influence on the body weight of mice after im-munization. Conclusion WCDSP and CCDPS could significantly enhance the OVA-specific humoral and cellular immune responses with no statistical difference and are characterized by high safety.
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This study sought to shed light on the killing effect of peripheral blood mononuclear cells (PBMCs) irradiated by gamma ray at a dose of 1 Gy on cultured human gastric tumor cell line MKN-28. The radiation dose rate of 17 Gy/min was used. The groups in the experiment were MKN-28 cell control group, PBMCs control group, MKN-28 tumor cells with irradiated or non-irradiated PBMCs co-cultured groups. Radiation dosage was one Gray, acridine orange/ethidium bromide (AO/EB) staining was used for observation of the killing effects of PBMCs on tumor cells in different period. Cells were harvested 240 h later and observed by transmission electron microscopy. The result showed the living period of irradiated PBMCs was shorter than that of non-irradiated PBMCs. In the irradiated and non-irradiated groups,a few PBMCs were still alive after being cultured for 240 h, but the cell volume was larger than that of lymphocytes. These cells were identified as monocytes (95%) or DCs (5%) by transmission electron microscopy. The co-culture of irradiated PBMCs and MKN-28 cells showed that tumor cells were eliminated after 96 h. As compared with the non-irradiated goup, the irradiated PBMCs had more potent ability for killing tumor. The results demonstrate that 1 Gy gamma irridiation can improve the killing effect of PBMCs on the tumor cells, and that 1 Gy gamma irritation can also induce shorter living period of lymphocytes in PBMCs cultured in vitro, but such irritation has little effect on the living period of monocytes and DCs in PBMCs.
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Humans , Cell Survival , Coculture Techniques , Gamma Rays , Leukocytes, Mononuclear , Cell Biology , Allergy and Immunology , Radiation Effects , Stomach Neoplasms , Allergy and Immunology , Pathology , Tumor Cells, CulturedABSTRACT
BACKGROUND:Previous animal studies have revealed that osteogenic growth peptide (OGP) applied locally or systemically could promote fracture healing. But the disadvantages of short in vivo half-life and low oral bioavailability limit its clinical application.OBJECTIVE: To study the encapsulation and delivery of synthetic OGP (sOGP) from biodegradable polymeric microspheres in vitro so as to choose better carrier for the future study.DESIGN: Grouping observation and comparative trail.SETTING: Laboratory of School of Life Science and Technology, Xi'an Jiaotong University.MATERIALS: sOGP was synthesized by Xi'an Langene Bio-science Co., Ltd. with Fmoc system. The purity of sOGP after purification was over 98 % identified by reverse phase high performance liquid chromatography, and the molecular weight of sOGP was 1 523 650, which was consistent with the theoretical value (Mr 1 523 750); the result of whole sequence analysis of sOGP was consistent with the theoretical sequence of OGP. Poly (lactic-co-glycolic acid) (PLGA,50:50, Mr 30 000; 75:25 Mr 80 000) was obtained from Shandong Medical Instrumental Institute (Ji'nan, China)METHODS: PLGA with a 50:50 or 75:25 lactide to glycolide ratio was used for microsphere preparation using a modified double emulsion solvent extraction Water-in-oil-in-water (w/o/w) technique. The surface structure and appearance of microsphere was observed under scanning electron microscope; particle size distribution of microsphere was counted by laser diffraction particle sizer; efficiency of encapsulation, release time and the structural integrity of sOGP released from PLGA were assessed using high performance liquid chromatography (HPLC).RESULTS: ①Spherical microspheres of sOG-PLGA were formulated successfully. The average particle diameter of the PLGA 50:50 microsphere was (19.6±4.5) μm, efficiency of encapsulation (83.9±4.2)% with (83.9±4.2) % drug-loading efficiency, while the PLGA 75:25 microspheres showed an average size of (35.8±3.6) μm, efficiency of encapsulation (65.6±6.8)% with (65.6±6.8)% drug-loading efficiency. ②HPLC results indicated that sOGP were not chemically altered,physically aggregated but presented a intact structure as the original sOGP. An initial burst release was observed for both PLGA microspheres, especially from PLGA 75:25. sOGP was released from PLGA 50:50 microsphere for 56 days,and from PLGA 75:25 microspheres for over 70 days. The cumulative release of sOGP from PLGA 50:50 for 35 days was significantly lower than from PLGA 75:25 (P ＜ 0.05).CONCLUSION: The controlled release of sOGP encapsulated within PLGA 50:50 is better than the delivery from PLGA 75:25. Moreover, the release time could meet the requirements for fracture or bone defect site.
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Objectives:To study the preparation techniques of gelatin microspheres encapsulated with sodium fluoride and evaluate the anticaries functions in dental plaque model in vitro. Methods:Fluoride gelatin microspheres were prepared by emulsion polymerisation method. The encapsulation rate and drug contents were tested by ISE. In vitro inhibition of demineralization were also examined. Results:The mean size of fluoride GMS was (16.81?8.77)?m.The encapsulation rate was 76.73%.The drug content was 5.61%.The release profile in vitro showed sustained effects. Fluoride GMS can prevent demineralization more efficiently. Conclusions:Fluoride gelatin microspheres may be an promising topical fluoride release system.
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Pingyangmycin gelatin microspheres(PYM-GMS) was prepared by optimal double-phase emulsified condensation polymerization for the interventional Chemoembolization with carotid artery therapy, and its release characteristics were studied in vivo and in vitro. The results of three ways of administration(vein drop, artery perfusion and artery embolization) were compared. The experiment indicates that the diameter of PYM-GMS is more appropriate for the application in external carotid artery embolization with PYM-GMS, which significantly reduces the circulating drug level and the dosage, prolongs the time period of higher drug concentration, achieves the purpose of sustained release and targeted tumor therapy.
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Animals , Female , Male , Rabbits , Antineoplastic Agents , Pharmacokinetics , Bleomycin , Pharmacokinetics , Chemotherapy, Cancer, Regional Perfusion , Drug Compounding , Methods , Gelatin , In Vitro Techniques , Infusions, Intravenous , Microspheres , Particle SizeABSTRACT
In this symposium, we reviewed the principle and development of time-resolved fluorescence anisotropy measurement. Its method of measurement, characteristics and applications in the research of biomacromolecule, configuration and molecular structure have been discussed. Its potential applications are also illustrated.
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Fluorescence Polarization , Methods , Macromolecular Substances , Mathematics , Models, Molecular , Time FactorsABSTRACT
This paper reviews recent research and development on microwave sterilization, including the experiments of sterilization effects, the influence of microwave radiating on the media parameters, the development of microwave sterilizers and the study of sterilization mechanism. Many results show that the method of microwave sterilization is more easy-to-perform, economical and reproducible than the conventional autoclaving method. It is necessary to study the sterilization conditions through the experiments for different media and bacteria. The research on sterilization mechanism, especially non-thermal sterilization mechanism is still a hotpoint.
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Bacteria , Radiation Effects , Microwaves , Research , Sterilization , MethodsABSTRACT
0.05). Conclusion After heated, the physical stability of UAE and UAS is reduced, the viscosity become lower, ADM releasing rate is fell. The heated Lipiodol-Adriamycin pharmaceutics had advantage in the interventional embolization chemotherapy of the neoplasm.
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Object To investigate the ultrafine grinding and nanometer powder's antiobesity effect of Amorphophallus rivieri Durieu on nutritional obese rats. Methods The content of triglyceride, cholesterol and high density lipoprotein (HDL) of blood was measured by UV spectraphotometry method. The blood glucose was determined by spectraphotometry method. The antiobesity effect was determined by weight loss and Lee's coefficient. Results Compared with powder and polysaccharide of A. rivieri, the ultrafine grinding and nanometer powder of A. rivieri could significantly decrease the rat body weight and the Lee's coefficient of nutritional obese rats (P
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Objective To evaluate the transfection efficiency and cytotoxicity of polyamine cationic liposomes (PCL), and to screen for a cationic liposome with high potency and low toxicity. Methods Four kinds of PCLs, namely PCL-A, PCL-B, PCL-C and PCL-D, were prepared with polyamine cationic lipid TC-Chol or DC-Chol and the neutral phospholipid DOPE in different molar ratios (1∶1 or 3∶1). The morphology of the liposome was observed by transmission electron microscope. Plasmid PIRES2-EGFP, which contained an enhanced green fluorescencent-protein (EGFP) reporter gene, was transfected into Hela cells or Hep2 cells with PCL-A, PCL-B, PCL-C and PCL-D, respectively. Expression of EGFP was examined by fluorescence microscopy and the cytotoxicity of the transfected cells was determined with MTT assay. Results The shape of PCL-A, PCL-B, PCL-C and PCL-D was round or oval, most of the PCLs were about 50-200nm in diameter. The transfection efficiency of the four kinds of PCLs was about 19.8%-42.1%. Among them, PCL-C prepared with TC-Chol and DOPE at a molar ratio of 3∶1 showed a higher transfection efficiency (39.5%-42.1%) and lower cytotoxicity (cellular survival rate was 86.57%?3.23%-84.43%?4.18%). Conclusions The polyamine cationic liposome prepared with polyamine cationic lipids TC-Chol or DC-Chol and the neutral phospholipid DOPE at a certain molar ratio mediated the transfection of the plasmid PIRES2-EGFP into mammalian cells. PCL-C exhibited higher transfection efficiency and lower cytotoxicity, and it might be a promising agent for gene transfection and gene therapy.