BACKGROUND: Warfarin is a widely used oral agent for anticoagulation therapy. Warfarin has a narrow therapeutic index and a wide variation in the interindividual therapeutic dosage. Recently, genotypes of CYP2C9 and VKORC1 have been found to account for 30-40% of the warfarin dosing variability, and a variety of commercial genotyping assays are being introduced. In this study, we evaluated the Verigene Warfarin Metabolism Nucleic Acid test (Verigene Warfarin assay; Nanosphere, USA) for its accuracy and clinical utility in genotyping CYP2C9*2, CYP2C9*3, and VKORC1 1173C>T. METHODS: We compared the Verigene Warfarin assay with direct sequencing for accuracy in determining the genotypes of CYP2C9*2, CYP2C9*3, and VKORC1 1173C>T using 50 patient samples and 3 commercial DNA samples with known genotypes. The method was also evaluated for turn-around time, hands-on time, and feasibility. RESULTS: The Verigene Warfarin assay demonstrated 100% accuracy for identifying CYP2C9*2, CYP2C9*3, and VKORC1 1173C>T. The turn-around time and hands-on time were 3 hr and 2 min, respectively. The no-call error rate at first attempt was estimated to be 2%. CONCLUSIONS: The Verigene Warfarin assay provides rapid and accurate genotype results. Considering there are only a few steps requiring manual intervention, it would be feasible to implement this assay even in clinical laboratories that lack considerable expertise in molecular diagnostics.
DNA ; Genotype ; Humans ; Metabolism* ; Nanospheres ; Pathology, Molecular ; Warfarin*

OBJECTIVETo prepare flexible proanthocyanidins nanoliposomes, and explore the in vitro release behavior of proanthocyanidins flexible nanoliposomes and general nanoliposomes.

METHODFlexible proanthoeyanidins nanoliposomes were prepared proanthocyanidins using a film dispersion method, characterized by transmission electron microscope, and the in vitro release action was studied in different dissolution mediums using dynamic dialyse method with the content of total phenol as index.

RESULTThe in vitro release of both proanthocyanidins flexible nanoliposomes and general nanoliposomes were in accordance with Weibull distribution.

CONCLUSIONProanthocyanidins flexible nanoliposomes without pressure had similar in vitro release behavior with general nanoliposomes.

Drug Delivery Systems ; methods ; Liposomes ; chemistry ; ultrastructure ; Nanospheres ; chemistry ; ultrastructure ; Particle Size ; Proanthocyanidins ; chemistry

Acute Lung Injury ; chemically induced ; pathology ; Animals ; Humans ; Lung ; pathology ; Nanospheres ; adverse effects ; Titanium ; adverse effects

Magnetic chitosan (CS) nano-spheres were prepared by the modified suspension cross-linking technique. The results demonstrated that the magnetic drug nano-spheres are mainly spherical in form with a size of 200 to 800 nm, and show good magnetic responsivity. Here, Doxorubicin was used as exam drug. Glutaraldehyde connects Doxorubicin to CS by the chemical bond (-N = C-), and the drug content is in range of 1% to 15% (w/w). The chemical bond is broken depending on pH, so pH is the important factor for the release of doxorubicin. The doxorubicin release was 22.0%, 13.4%, and 4.1% in the space of 7d, when pH was 1, 2, 4. So the nano-spheres are pH-sensitive magnetic targeting drug micro-spheres.
Chitosan ; chemistry ; Doxorubicin ; administration & dosage ; Drug Carriers ; chemical synthesis ; Drug Delivery Systems ; methods ; Magnetics ; Nanospheres ; chemistry

Objective To investigate the anti-inflammatory effect of artemisinin (ART) encapsulated by β-lactoglobulin (BLG) nanoparticles on Winnie spontaneous ulcerative colitis mouse model. Methods BLG-ART nanoparticles were prepared and their effects on the solubility and stability of ART were evaluated. A mouse model of colitis induced by dextran sulfate sodium (DSS) was used to compare the therapeutic effects of artemisinin (ART) administered by direct gavage and artemisinin encapsulated by β-lactoglobulin nanoparticles (BLG-ART) administered by gavage. Winnie mice were randomly divided into blank group, ART group and BLG-ART group. Mice in the ART group were given 50 mg/kg ART by gavage; mice in the BLG-ART group were given the same dose of BLG-ART nanoparticle PBS dispersion by gavage; mice in the blank group were given the same amount of PBS by gavage, for 16 days. The body mass and disease activity index (DAI) of each group of mice were measured. HE staining was used to observe the pathological changes of mouse intestinal tissue, and real-time quantitative PCR was used to detect the mRNA expression levels of TNF-α, interleukin 1β (IL-1β), IL-10 and IL-17 in mouse colon tissue. Results Compared with the ART group and the blank group, the body mass of the BLG-ART group increased and the DAI decreased after 16-day treatment; the crypt structure of the proximal and distal colon regions of the mice recovered; goblet cell loss decreased; neutrophil infiltration decreased and the mRNA expression levels of pro-inflammatory and anti-inflammatory cytokines were significantly down-regulated. Conclusion ART-BLG can alleviate intestinal inflammation in spontaneous ulcerative colitis mice.
Animals ; Mice ; Colitis, Ulcerative/drug therapy* ; Nanospheres ; Inflammation ; Administration, Oral ; Artemisinins ; Disease Models, Animal ; RNA, Messenger

OBJECTIVETo investigate the effect of modified bone morphogenetic protein-2 polylactic acid nanospheres (BMP-2-PLA-Ns) sustained-release system on rabbit mandibular defect repair.

METHODSThe polylactic acid s nanospheres (PLA-Ns) and BMP-2-PLA-Ns were prepared by ultrasonic emulsification after graft polymerization. Forty-five rabbits were randomly divided into 3 groups: Blank group, PLA-Ns gel group(control group), and BMP-2-PLA-Ns gel group (experimental group). The rabbit mandibular defect models were established. The defect area of control group was implanted with PLA-Ns gel, meanwhile, the experimental group was implanted with BMP-2-PLA-Ns gel, the blank group experienced no special handling. Rabbits were killed in 1, 2, 4 weeks after operation and the iconography, hematine eosin(HE) staining and PCNA immunohistochemistry were used to detect the reparative effect on rabbit mandible defects.

RESULTSImage observation showed that bone defect repair in the experimental group was well and the shadow was not obvious. Better repair effect was seen compared with the control group and blank group. HE staining showed that the experimental group and the control group had a large number of neovascularization and secondary callus formation, callus in experimental group was obviously higher than that of control group and blank group. Immunohistochemical observation showed that the experimental group's PCNA positive chondrocytes were more than those in the control group and the blank group in the first 2 weeks; all groups of PCNA positive cells were rare in the fourth week, PCNA positive expression rate of the fourth week was lower than that of the first 2 weeks.

CONCLUSIONThe modified BMP-2-PLA-Ns sustained-release system promotes mandibular defect repair obviously.

Animals ; Bone Morphogenetic Protein 2 ; Delayed-Action Preparations ; Lactic Acid ; Mandible ; Nanospheres ; Polyesters ; Polymers ; Rabbits ; Reconstructive Surgical Procedures

BACKGROUND: Currently, dermal fillers need to be 25 µm or larger to reduce in vivo degradation by macrophages. However, the large size of fillers may cause side effects, including interruption of blood flow and nodule formation. Therefore, using rats, we tested a polycaprolactone copolymer hydrogel with nanoscale particles that could maintain a low in vivo degradation rate. METHODS: Thirty-six 6-week-old Sprague-Dawley rats were divided into group A (normal saline), group B (polycaprolactone microsphere filler), and group C (polycaprolactone copolymer nanosphere hydrogel). The corresponding materials were injected into the dermal layer of the scalp of the rats. At 4, 8, and 12 weeks after injection, blood biochemical and kidney and liver histological analyses were performed. Tissues were examined using hematoxylin-eosin staining to observe tissue infiltration of materials. Collagen formation in the dermal tissue of the scalp was observed with Masson trichrome staining and the collagen content was quantified using a soluble collagen assay kit. RESULTS: The histologic examination for organ infiltration showed no abnormal findings. All blood test results were within the normal ranges. The amount of collagen at 12 weeks increased by 1.22 mg/g in group C and by 0.6 mg/g in group B. CONCLUSIONS: The results reveal that the nanosphere complex near the injection site induced collagen formation. Regardless of the sphere size, aggregation of the copolymer prevented macrophage phagocytosis. The polycaprolactone copolymer nanosphere hydrogel was effective for more than 3 months when injected in the scalp dermal tissue of Sprague-Dawley rats and can be used safely.
Animals ; Collagen ; Dermal Fillers ; Hematologic Tests ; Hydrogel ; Kidney ; Liver ; Macrophages ; Microspheres ; Nanospheres ; Phagocytosis ; Rats ; Rats, Sprague-Dawley ; Reference Values ; Scalp

A 69-year-old female Korean patient was initially prescribed warfarin for the prevention of systemic thromboembolism due to atrial fibrillation. One month later, multiple bruises and subcutaneous hematomas were evident, and laboratory testing revealed a prolonged prothrombin time (PT) of > 106s. After admission, the PT was corrected via fresh frozen plasma transfusion and intravenous vitamin K infusion. We sought to determine the cause of the PT prolongation, suspecting that genetic cause may have had an effect on the variation in the warfarin dose requirement. A point-of-care gene test device (Verigene(R) system; Nanosphere, Northbrook, IL) revealed CYP2C9*1/*3 heterozygosity and a VKORC1 A/A single nucleotide polymorphism. Although it is well established that CYP2C9 or VKORC1 gene polymorphisms can influence warfarin dose requirements, they can be easily neglected, with detrimental outcomes. Through our experience with CYP2C9 and VKORC1 polymorphism causing bleeding complications during warfarin treatment, we aim to emphasize the importance of pharmacogenetic testing to avoid this potential oversight.
Atrial Fibrillation ; Contusions ; Female ; Hematoma ; Hemorrhage ; Humans ; Nanospheres ; Pharmacogenetics ; Plasma ; Point-of-Care Systems ; Polymorphism, Single Nucleotide ; Prothrombin Time ; Thromboembolism ; Vitamin K ; Warfarin

Ampicillin sodium was embeded in ethylcellulose (EC) nanospheres made of low-molecular-weight EC. Low-molecular-weight EC was attained with ethylcellulose being degraded by 34% (w/w) nitric acid; Fourier transform infrared (FTIR) spectroscopy, 13C-NMR, element analysis confirmed that the basic structure and major properties of low-molecular-weight EC maintained agreement with those of undegraded EC except that the polymerization degree of EC decreased. Wide-angle X-ray diffraction demonstrated that the crystallinity of degraded EC decreased. Ampicillin sodium loaded EC nanospheres were characterized by transmission electron microscopy, FTIR and in vitro drug release. Molecular weight of EC would affect the size of nanospheres, distribution and drug encapsulation efficiency. Drug loaded nanospheres resulted in the drug control release in 3-10 hours.
Ampicillin ; administration & dosage ; chemistry ; Anti-Bacterial Agents ; administration & dosage ; chemistry ; Cellulose ; administration & dosage ; analogs & derivatives ; chemistry ; Delayed-Action Preparations ; chemical synthesis ; Nanospheres ; Nanotechnology ; methods ; Spectroscopy, Fourier Transform Infrared ; X-Ray Diffraction

OBJECTIVETo prepare nanospheres coupled with the anti-human liver cancer monoclonal antibody HAb18 and evaluate its immunoreactivity and antitumor effects.

METHODSThe nanosphere coupled with the antibody was prepared by intermolecular cross-linking the anti-human liver cancer monoclonal antibody, HAb18, with human serum albumin nanospheres containing ADM [termed HAS(ADM)-NS] via a new hetero-bifunctional cross-linker SPDP. Condensation test and immunofluorescence assay were used to evaluate the immunoreactivity of the nanospheres, and specific binding of HAb18-HAS(ADM)-NS with liver cancer cell line SMMC-7721 was observed with optical and electron microscopes. The specific cytotoxic effects on the target cells were evaluated in vitro by MTT assay. HAb18-HAS(ADM)-NS, HAS(ADM)-NS and ADM were injected separately into nude mice bearing human liver carcinoma to evaluate the inhibitory activity of HAb18-HAS(ADM)-NS in vivo.

RESULTSThe immunoreactivity of HAb18-HAS(ADM)-NS was well preserved. HAb18-HAS(ADM)-NS could bind specifically with the SMMC-7721 cells. The IC(50) of HAb18-HAS(ADM)-NS against SMMC-7721 cells was 44.6 microg/ml, lower than that of HAS(ADM)-NS (345.5 microg/ml) and ADM (365.5 microg/ml). The inhibition rate of HAb18-HAS(ADM)-NS on the growth of liver cancer xenografts was significantly higher than that of HAS(ADM)-NS and ADM (P<0.001).

CONCLUSIONHAb18-HAS(ADM)-NS has immunoreactivity and can actively and specifically target the liver cancer cells. The antitumor activity of HAb18-HAS(ADM)-NS is significantly higher than that of HAS(ADM)-NS and ADM.

Animals ; Antibodies, Monoclonal ; administration & dosage ; immunology ; Antibodies, Neoplasm ; immunology ; Antineoplastic Combined Chemotherapy Protocols ; immunology ; therapeutic use ; Cell Line, Tumor ; Doxorubicin ; administration & dosage ; immunology ; Female ; Humans ; Immunotoxins ; administration & dosage ; immunology ; Liver Neoplasms ; drug therapy ; immunology ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Nanospheres ; administration & dosage ; Treatment Outcome ; Xenograft Model Antitumor Assays ; methods