OBJECTIVETo prepare a platinum microcoil coated with polymers and vascular endothelial growth factor (VEGF), and evaluate its surface characteristics and property of sustained VEGF release.

METHODSThe surface of the platinum microcoils (GDC) were modified by coating P(DLLA-co-TMC) copolymer and immobilizing heparin on the surface of GDC. VEGF was then loaded onto the surface of GDC and the controlled release of VEGF within GDC was achieved. The morphology was observed by scanning electron microscope, and the sustained release of VEGF was evaluated by enzyme-linked immunosorbent assay (ELISA).

RESULTSPlatinum coils were prepared by successive deposition of P(DLLA-co-TMC) copolymer and anionic heparin, and VEGF was immobilized through affinity interaction with heparin. The accumulative release of VEGF increased obviously during the entire testing period without burst release.

CONCLUSIONThe use of P(DLLA-co-TMC) copolymer allows immobilization of VEGF on the platinum coils for controlled VEGF release, and improves the biological property of the coils.

Coated Materials, Biocompatible ; chemistry ; Delayed-Action Preparations ; pharmacology ; Platinum ; chemistry ; Polymers ; chemistry ; Vascular Endothelial Growth Factor A ; pharmacology

Objective To prepare PLGA/PEG microsphcres for sustained release of bovine serum albumin (BSA), and evaluate the effect of the micrnsphere composition, proportion of the components and integration of PEG on the size, BSA encapsulation rate and in vitro BSA release properties of the microspheres. Methods BSA-loaded PLGA/PEG microspheres were prepared by water-in-oil-in-water double emulsions-solvent evaporation method (W/O/W). The morphology of the microspheres was observed with scanning elect'on microscope, the diameter was determined by Mastersizer 2000, and the encapsulation efficiency and in vitro BSA release were evaluated by UV spectrophotometry. Results Increased GA to LA ratio was associated with increased size of the microspheres and accelerated BSA release. The incorporation of PEG obviously increased the BSA encapsulation rate to as much as 88.22% with also increased BSA release rate. The BSA-PLGA/PEG biodegradable microspheres prepared was characterized by high encapsulation rate, low burst release, and slow BSA release for over 3 weeks at high drug concentrations. Conclusion BSA-PLGA/PEG microspheres with relatively high encapsulation efficiency and proper drug loading can be prepared by adjusting the parameters during the preparation process.

The prokaryotic expression plasmid pET-30a(+)/sBLyS was constructed and transformed into E. coli BL21 (lambda DE3). The recombinant protein was found to be highly expressed by the plight of soluble part and inclusion body. For the sake of enhancing the proportion of the soluble part, inducement at 16 degrees C for 12 h was ascertained. The expressing product was then purified by Ni2+ affinity chromatography gel. PI of the recombinant human sBLyS(rhsBLyS) is about 7.1-7.3 and it assembles into a homotrimer. The effect of rhsBLyS on B lymphocytes by MTT method told us the B lymphocytes' proliferating capacity dose depended on concentration and also stimulating time of the rhsBLys. With rhsBLyS(2 micrograms/mL) stimulating 3 days, B lymphocytes can proliferate the most.
B-Cell Activating Factor ; B-Lymphocytes ; drug effects ; Escherichia coli ; genetics ; Humans ; Isoelectric Point ; Lymphocyte Activation ; drug effects ; Membrane Proteins ; biosynthesis ; isolation & purification ; pharmacology ; Recombinant Proteins ; biosynthesis ; Temperature ; Tumor Necrosis Factor-alpha ; biosynthesis ; isolation & purification ; pharmacology

The chitosan/PHEA-blended hydrogels were prepared from PHEA and chitosan in various blend ratios. The water contents of the hydrogels were in the range of 50%-80% (wt). The attachment and growth of fibroblast cells(L929) on the hydrogels were studied. The results indicated the PHEA content in hydrogels has great effect on cell attachment but has little effect on the growth of L929 cells.
Animals ; Biocompatible Materials ; chemistry ; Cell Adhesion ; physiology ; Cell Division ; physiology ; Cells, Cultured ; Chitin ; analogs & derivatives ; chemistry ; Chitosan ; Fibroblasts ; cytology ; physiology ; Hydrogel, Polyethylene Glycol Dimethacrylate ; chemistry ; Mice ; Peptides ; chemistry ; Water ; chemistry

OBJECTIVETo evaluate the clinical outcome of periodontal initial therapy on the patients with aggressive periodontitis.

METHODSA total of 48 patients with aggressive periodontitis, including 20 patients with localized aggressive periodontitis (LAgP) and 28 patients with generalized aggressive periodontitis (GAgP), were chosen for the study from the patients referred to the Department of Periodontology of the School of Stomatology of China Medical University from September 2006 to December 2008. All subjects were performed periodontal initial therapy. Probing depth (PD), clinical attachment loss (CAL), bleeding index (BI) and tooth mobility were recorded at baseline and 1, 3 and 6 months after initial treatment.

RESULTSAt 1, 3 and 6 months after periodontal initial therapy, there were significant improvements in PD, CAL, BI and tooth mobility of patients with LAgP and GAgP than those in the baseline (P < 0.05). In patients with LAgP, statistical differences were detected in PD and CAL between 1 and 3 months after periodontal initial therapy (P < 0.05). And in patients with GAgP, statistical differences were detected in PD and CAL between 3 and 6 months after therapy (P < 0.05). At 1, 3, and 6 months after therapy, first molars of all patients showed more significant therapeutic effects than central incisors (P < 0.05).

CONCLUSIONPeriodontal initial therapy showed effectiveness in treatment of aggressive periodontitis. In the mid-term, there were differences in therapeutic effect between patients with LAgP and GAgP.

Adult ; Aggressive Periodontitis ; China ; Female ; Humans ; Male ; Periodontitis

OBJECTIVE: To investigate the relationshion between the angiogenesis of different kinds of scar and expression of HO-1. METHODS: The expression of heme oxygenase-1 and vessel counted by CD34 of biopsies from different kinds of scars such as hypertrophic scar, keloid, surgical scar and normal skin of 24 cases was valued by immunochemical method, and the relationship was compared between them. RESULTS: The vessel count of hypertrophic scar, keloid was significantly abundant compared with surgical scar or normal skin (P < 0.01). While the expression of HO-1 of hypertrophic scar, keloid was obviously higher than that in surgical scar or normal skin (P < 0.01), decreased from hypertrophic scar, keloid, surgical scar to normal skin. There existed a positive correlation between vessel count and the expression of HO-1 (r = 0. 761, P < 0.01) as well as the number of fibroblastic cells (r = 0. 731, P < 0.01) in the study groups. CONCLUSION HO-1 might play a important role in the angiogenesis of scar formation. The cause of these changes may be local. Over angiogenesis is one symbol of pathological scar.
Adult ; Cicatrix ; metabolism ; pathology ; Cicatrix, Hypertrophic ; metabolism ; pathology ; Female ; Heme Oxygenase-1 ; biosynthesis ; genetics ; Humans ; Keloid ; metabolism ; pathology ; Male ; Middle Aged ; Neovascularization, Pathologic ; Skin ; blood supply

OBJECTIVETo assess the feasibility and safety of radiofrequency ablation (RFA) in spleen to treat secondary splenomegaly and hypersplenism in dogs.

METHODSFourteen healthy mongrel dogs were randomly divided into two groups: group A (n = 4) and group B (n = 10) Both groups underwent ligation of the splenic vein and its collateral branches to induce congestive splenomegaly. At the end of the 3rd week, radiofrequency thermal ablation of the spleen was performed in the group B via laparotomy. After splenic RFA, the procedure-related complications were observed, CT scan was performed and the spleens were obtained according to schedule. The radiofrequency (RF) thermal lesions and its histo- pathological changes of the spleen were examined regularly.

RESULTSThere were no morbidity and mortality in the experimental dogs. CT findings revealed that splenomegaly could sustained over 2 months after ligation of the splenic vein. The segmental RF lesions included hyperintense zone of coagulative necrosis and more extensively peripheral hypo-intense infarcted zone. The latter was called as "bystander effect". The infarcted zone would be absorbed and subsequently disappeared between 4 and 6 weeks after RFA, and the size of the remnant spleen shrunk, but the lesion of coagulative necrosis hardly altered. The fundamental histopathological changes of splenic lesions caused by RF thermal energy included local coagulative necrosis and the peripheral thrombotic infarction zone. Subsequently, tissue absorption and fibrosis occurred in the zone of thrombotic infarction. Simultaneously occluded vessels, fibrin deposition, and disappearance of normal splenic sinuses resulted in the condensed structure of the viable remnant spleen, which were the pathological basis responsible for the shrunk spleen.

CONCLUSIONSIt is feasible and safe to perform RFA in the spleen to treat experimental splenomegaly and hypersplenism. The RFA technique could be safely performed clinically via laparotomy or laparoscopic procedure to strictly isolate the spleen from the surrounding organs.

Animals ; Catheter Ablation ; Disease Models, Animal ; Dogs ; Feasibility Studies ; Female ; Hypersplenism ; etiology ; pathology ; surgery ; Ligation ; adverse effects ; Male ; Random Allocation ; Spleen ; diagnostic imaging ; pathology ; Splenectomy ; methods ; Splenic Vein ; surgery ; Splenomegaly ; etiology ; pathology ; surgery ; Tomography, X-Ray Computed

OBJECTIVE: To explore the application value of serum total IgE, tryptase and chymase in the identification of death caused by drug anaphylactic shock. METHODS: The general information from 235 cases of non-drug anaphylactic shock and 32 cases of drug anaphylactic shock were analyzed. The serum IgE level had been detected in the cases. Ten cases caused by coronary disease and 10 cases caused by sudden manhood death syndrome were selected from non-drug anaphylactic shock cases for the control group. Expressions of tryptase and chymase in the lung and heart were detected using immunohistochemistry method. The number and IOD of positive mast cells were counted. RESULTS: In the drug anaphylactic shock group, the IgE value of 18 samples (56.25%) was significantly higher than the normal upper limit of 120 IU/mL. In the non-drug anaphylactic shock group, the IgE value of 67 samples (28.51%) was higher than 120 IU/mL. The expressions of tryptase and chymase were significantly increased in lung and myocardial tissue in drug anaphylactic shock group (P < 0.05). CONCLUSION Tryptase and chymase are more superior than that of the serum total IgE in the diagnosis of death caused by drug anaphylactic shock, and are more suitable in forensic practice.
Adolescent ; Adult ; Aged ; Anaphylaxis/pathology* ; Autopsy ; Case-Control Studies ; Cause of Death ; Child ; Child, Preschool ; Chymases/metabolism* ; Death, Sudden, Cardiac/pathology* ; Drug Hypersensitivity ; Female ; Forensic Pathology ; Humans ; Immunoglobulin E/blood* ; Immunohistochemistry ; Infant ; Lung/pathology* ; Male ; Middle Aged ; Myocardium/pathology* ; Tryptases/metabolism* ; Young Adult

BACKGROUNDNatural articular cartilage has a limited capacity for spontaneous regeneration. Controlled release of transforming growth factor-beta1 (TGF-beta1) to cartilage defects can enhance chondrogenesis. In this study, we assessed the feasibility of using biodegradable chitosan microspheres as carriers for controlled TGF-beta1 delivery and the effect of released TGF-beta1 on the chondrogenic potential of chondrocytes.

METHODSChitosan scaffolds and chitosan microspheres loaded with TGF-beta1 were prepared by the freeze-drying and the emulsion-crosslinking method respectively. In vitro drug release kinetics, as measured by enzyme-linked immunosorbent assay, was monitored for 7 days. Lysozyme degradation was performed for 4 weeks to detect in vitro degradability of the scaffolds and the microspheres. Rabbit chondrocytes were seeded on the scaffolds containing TGF-beta1 microspheres and incubated in vitro for 3 weeks. Histological examination and type II collagen immunohistochemical staining was performed to evaluate the effects of released TGF-beta1 on cell adhesivity, proliferation and synthesis of the extracellular matrix.

RESULTSTGF-beta1 was encapsulated into chitosan microspheres and the encapsulation efficiency of TGF-beta1 was high (90.1%). During 4 weeks of incubation in lysozyme solution for in vitro degradation, the mass of both the scaffolds and the microspheres decreased continuously and significant morphological changes was noticed. From the release experiments, it was found that TGF-beta1 could be released from the microspheres in a multiphasic fashion including an initial burst phase, a slow linear release phase and a plateau phase. The release amount of TGF-beta1 was 37.4%, 50.7%, 61.3%, and 63.5% for 1, 3, 5, and 7 days respectively. At 21 days after cultivation, type II collagen immunohistochemical staining was performed. The mean percentage of positive cells for collagen type II in control group (32.7% +/- 10.4%) was significantly lower than that in the controlled TGF-beta1 release group (92.4% +/- 4.8%, P < 0.05). Both the proliferation rate and production of collagen type II in the transforming growth factor-beta1 microsphere incorporated scaffolds were significantly higher than those in the scaffolds without microspheres, indicating that the activity of TGF-beta1 was retained during microsphere fabrication and after growth factor release.

CONCLUSIONChitosan microspheres can serve as delivery vehicles for controlled release of TGF-beta1, and the released growth factor can augment chondrocytes proliferation and synthesis of extracellular matrix. Chitosan scaffolds incorporated with chitosan microspheres loaded with TGF-beta1 possess a promising potential to be applied for controlled cytokine delivery and cartilage tissue engineering.

Animals ; Cartilage ; metabolism ; Cell Proliferation ; Chitosan ; administration & dosage ; Chondrocytes ; cytology ; Drug Carriers ; Microspheres ; Rabbits ; Tissue Engineering ; methods ; Transforming Growth Factor beta1 ; administration & dosage ; chemistry

This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Base Sequence ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Lentivirus ; genetics ; metabolism ; Molecular Sequence Data ; RNA Interference ; RNA, Small Interfering ; genetics ; metabolism ; Receptors, KIR ; biosynthesis ; genetics