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Sheng S.,Liaoning Medical University | Kong F.,General Hospital of Jinan Command
Pharmaceutical Biology | Year: 2012

Context: Affinity chromatography is an efficient antibody, antigen and protein separation method based on the interaction between specific immobilized ligands and target antibody, antigen, and so on. Populations of available ligands can be used to separate antibodies or their Fab fragments. Similarly, antigens can be isolated by immunoaffinity chromatography (IAC) on immobilized antibodies of low affinity. Objective: This review describes the advantages, the applications, as well as the drawbacks, of IAC in the separation and purification of antibodies and antigens. Methods: The present review discussed all types of purification and isolation of antibodies and antigens by IAC, including purification of antibodies using immobilized and synthetic mimic proteins A, G and L; isolation of Fab fragments of antibodies; separation of antibodies against different antigen forms; isolation of antigens by immobilized antibodies and so on. These methods come from over 60 references compiled from all major databases. Results: Purification of antigens with antibodies should choose low-affinity antibodies to avoid denaturation of most proteins. Concern for cost and safety, prompted research activities focused on novel synthetic ligands with improved properties such as lower cost, avoidance of the risk of contamination associated with natural ligands of human or animal origin to isolate antibodies and antigens. Conclusion: It is anticipated that the improvements of IAC will have impact not only on large-scale production of antibodies but also on the generation of new affinity-based methods for the increasing number of proteins and antibody derivatives available by protein engineering and the proteomics revolution. © 2012 Informa Healthcare USA, Inc.


Zeng C.-H.,Nanjing University | Le W.,Nanjing University | Ni Z.,Shanghai JiaoTong University | Zhang M.,Shanghai JiaoTong University | And 40 more authors.
American Journal of Kidney Diseases | Year: 2012

Background: The Oxford classification of immunoglobulin A (IgA) nephropathy (IgAN) provides a histopathologic grading system that is associated with kidney disease outcomes independent of clinical features. We evaluated the Oxford IgAN classification in a large cohort of patients from China. Study Design: Retrospective study. Setting & Participants: 1,026 adults with IgAN from 18 referral centers in China. Inclusion criteria and statistical analysis were similar to the Oxford study. Predictors: Histologic findings of mesangial hypercellularity score, endocapillary proliferation, segmental sclerosis or adhesion, crescents, necrosis, and tubular atrophy/interstitial fibrosis. Clinical features, blood pressure, estimated glomerular filtration rate (eGFR), proteinuria, and treatment modalities. Outcomes: Time to a 50% reduction in eGFR or end-stage renal disease (the combined event); the rate of eGFR decline (slope of eGFR); proteinuria during follow-up. Results: Compared with the Oxford cohort, the Chinese cohort had a lower proportion of patients with mesangial hypercellularity (43%) and endocapillary proliferation (11%), higher proportion with segmental sclerosis or adhesion (83%) and necrosis (15%), and similar proportion with crescents (48%) and tubular atrophy/interstitial fibrosis (moderate, 24%; severe, 3.3%). During a median follow-up of 53 (25th-75th percentile, 36-67) months, 159 (15.5%) patients reached the combined event. Our study showed that patients with a mesangial hypercellularity score higher than 0.5 were associated with a 2.0-fold (95% CI, 1.5-2.8; P<0.001) higher risk of the combined event than patients with a score of 0.5 or lower. Patients with tubular atrophy/interstitial fibrosis of 25%-50% and >50% versus <25% were associated with a 3.7-fold (95% CI, 2.6-5.1; P<0.001) and 15.1-fold (95% CI, 9.5-24.2; P<0.001) higher risk of the combined event, respectively. Endocapillary proliferation, glomerular crescents, and necrosis were not significant. Limitations: Retrospective study; the therapeutic interventions were miscellaneous. Conclusions: We confirmed the associations of mesangial hypercellularity and tubular atrophy/interstitial fibrosis with kidney disease outcomes. © 2012 National Kidney Foundation, Inc.


Wang W.,Integrated Traditional Chinese Medicine and Western Medicine | Zhou F.,General Hospital of Jinan Command | Ge L.,General Hospital of Jinan Command | Liu X.,General Hospital of Jinan Command | Kong F.,General Hospital of Jinan Command
International Journal of Nanomedicine | Year: 2012

Background: The main barriers to non-viral gene delivery include cellular and nuclear membranes. As such, the aim of this study was to develop a type of vector that can target cells through receptor-mediated pathways and by using nuclear localization signal (NLS) to increase the nuclear uptake of genetic materials. Methods: A dexamethasone (Dexa)-conjugated lipid was synthesized as the material of the solid lipid nanoparticles (SLNs), and transferrin (Tf) was linked onto polyethylene glycolphosphatidylethanolamine (PEG-PE) to obtain Tf-PEG-PE ligands for the surface modification of the carriers. The in vitro transfection efficiency of the novel modified vectors was evaluated in human hepatoma carcinoma cell lines, and in vivo effects were observed in an animal model. Results: Tf-PEG-PE modified SLNs/enhanced green fluorescence protein plasmid (pEGFP) had a particle size of 222 nm and a gene loading quantity of 90%. Tf-PEG-PE-modified SLNs/ pEGFP (Tf-SLNs/pEGFP) displayed remarkably higher transfection efficiency than non-modified SLNs/pEGFP and the vectors not containing Dexa, both in vitro and in vivo. Conclusion: It can be concluded that Tf and Dexa could function as an excellent active targeting ligand to improve the cell targeting and nuclear targeting ability of the carriers, and the resulting nanomedicine could be a promising active targeting drug/gene delivery system. © 2012 Zeng et al, publisher and licensee Dove Medical Press Ltd.


Wang W.,Integrated Traditional Chinese Medicine and Western Medicine | Zhou F.,General Hospital of Jinan Command | Ge L.,General Hospital of Jinan Command | Liu X.,General Hospital of Jinan Command | Kong F.,General Hospital of Jinan Command
Pharmaceutical Biology | Year: 2014

Context: Non-viral gene delivery could deliver drugs/genes through cellular membranes and nuclear membranes by some modification of materials. Objective: This study develops a kind of vector to target the cells through receptor-mediated pathways. Nuclear localization signal (NLS) was also used to increase the nuclear uptake of genetic materials. Materials and methods: A lipid containing dexamethasone (Dexa) was synthesized as the material of the preparation of solid lipid nanoparticles (SLNs) and folate (Fa)-conjugated PEG-PE (Fa-PEG-PE) ligands were used to modify the SLNs. The in vitro cytotoxicity of the carriers at various concentrations (10, 20, 50, 100, and 200μg/ml) were evaluated in KB human carcinoma cells (KB cells). In vivo transfection efficiency of the novel modified vectors was evaluated in disseminated peritoneal tumors on mice bearing KB cells. Results: Fa-PEG-PE modified SLNs/enhanced green fluorescence protein plasmid (pEGFP) has a particle size of 258nm, and the gene loading quantity of the vector was 90%. The in vitro cytotoxicity of Fa-PEG-PE-modified SLNs/pEGFP (Fa-SLNs/pEGFP) was low (cell viabilities were between 80% and 100% compared with controls). Fa-SLNs/pEGFP displayed remarkably higher transfection efficiency (40%) than non-modified SLNs/pEGFP (24%) and the vectors not containing Dexa (30%) in vivo. Conclusion: The results demonstrate that Fa and Dexa could function as excellent active targeting ligands to improve the cell targeting and nuclear targeting ability of the carriers and the resulting vectors could be promising active targeting drug/gene delivery systems. © 2014 Informa Healthcare USA, Inc.


Tan H.,General Hospital of Jinan Command | Chen R.-M.,General Hospital of Jinan Command | Li X.-Y.,General Hospital of Jinan Command | Zhang H.-M.,General Hospital of Jinan Command | Yang Y.-Y.,Peoples Hospital of Guizhou Province
Medical Journal of Chinese People's Liberation Army | Year: 2014

Objective To investigate the effects of rosuvastatin on the expression profile of peripheral blood CD4+T lymphocytes miRNA gene in the patients with acute coronary syndrome (ACS) and screen the differentially expressed miRNAs before and after treatment, and elucidate the mechanisms of rosuvastatin calcium in the treatment of patients with ACS. Methods Nine cases were selected from the patients with ACS treated in the General Hospital of Jinan Military Command from Mar. to Jul. of 2012, with other 9 cases selected as controls, whose degree of coronary artery stenosis was less than 50% as confirmed by 320CT. Peripheral blood mononuclear cells were isolated by density gradient centrifugation, and CD4+lymphocytes were isolated by immunomagnetic beads method. miRNAs ot CD4+lymphocytes were detected by miKJNA gene chip technology. 1 he differentially expressed miRNAs between ACS patients and normal control, and those in ACS patients before and after treatment were screened. Three of the maximum difference miRNAs were selected and followed by verification by real-time polymerase chain reaction (RT-PCR). Results More than 1900 genes were detected by gene microarray, of which more than 300 genes showed significant differences in expression. Comparing the ACS patients and normal controls, 126 genes were significantly up-regulated, including miRNA-21, miRNA-142, and miRNA-20a; and 202 genes were significantly down-regulated, including miRNA-4734, miRNA-1182, and miRNA-1273f. A total of 157 genes were significantly up-regulated after treatment with rosuvastatin calcium (20mg/d×10d), such as miRNA-4734, miRNA-1182, and miRNA-663b; and 137 genes were significantly down-regulated, such as miRNA-4789, miRNA-S692c, and miRNA-26a. The results were validated by RT-PCR and they were same as miRNA microarray. Conclusion Rosuvastain may play a role in the treatment of patients with ACS by regulating a series of miRNAs such as miRNA-4734, miRNA-1182, and miRNA-4789.

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