AIMTo produce poly (lactic-co-glycolic acid) (PLGA) microspheres, containing a staphylokinase variant (K35R, DGR) with reduced immunogenecity and antiplatelet aggregation activities, which allowed the preservation of protein stability during both particle processing and drug release.

METHODSDGR-loaded microspheres were fabricated using a double emulsion-solvent evaporation technique. The effects of preparative parameters, such as stirring rate, polymer concentration, and the excipients of both internal and external aqueous phase (W2), on DGR encapsulation efficiency and microsphere characteristics were investigated. In vitro and in vivo release of DGR were conducted and the cause for instability of DGR during release was also investigated.

RESULTSModerate ultrasonic treatment of aqueous DGR/dichloromethane mixtures caused approximately. Eighty four per cent DGR denaturation. However, the activity recovery of DGR almost amounted to 100% when 2% polyvinyl alcohol (PVA) was addled into the aqueous phase. It was found that NaCl in the external water phase significantly increased DGR encapsulation efficiency. Furthermore, NaCl in the external water phase played a role in determining size and surface morphology of microsphere. In vitro release test showed a burst release of DGR from microspheres, followed by sustained release of 50% total activity over 15 days. In vivo experiments showed that DGR released from microspheres sustained 5 days. Denaturation of DGR within microspheres might be resulted from acidic microclimate.

CONCLUSIONThe stability of DGR was effectively protected during microencapsulation and a relatively high encapsulation efficiency of DGR was obtained. PLGA microspheres could be an effective carrier for DGR.

Animals ; Area Under Curve ; Drug Carriers ; Drug Compounding ; Drug Delivery Systems ; Escherichia coli Proteins ; administration & dosage ; genetics ; pharmacokinetics ; Genetic Variation ; Lactic Acid ; Male ; Metalloendopeptidases ; administration & dosage ; genetics ; pharmacokinetics ; Microspheres ; Particle Size ; Polyglycolic Acid ; Polymers ; Rabbits

Objective To study effective treatment for deep burn on the face caused by coal-dust burning and blasting to prevent severe disfigurement.Methods Early conservative eschar-scraping, delayed skin graft,early strain-diminution for the eyelids in both sides,and wound expansion with skin graft were used to treat 12 patients with deep burn on their faces caused by coal-dust burning and blasting. Results Facial wound of all the 12 patients healed within three weeks after burn.No significant cicatricial hyperplasia and deformity were found on their faces during three-month follow-up,with natural facial expression and abundant emotion.Conclusions Early eschar-scraping and delayed skin graft for deep facial burn can promote fast repair of burn-wound,diminish cicatricial hyperplasia and prevent deformity on the face.

Objective To explore the effect of dendritic cells (DC) modified with transforming growth factor β1 (TGF-β1) gene on the expressions of CD28/CTLA-4:B7 costimulatory molecules in peripheral blood mononuclear cells (PBMC) in the Lewis rats with experimental autoimmune myasthenia gravis (EAMG). Methods Thirty inbreeding line, healthy, female Lewis rats were divided randomly into 6 groups: normal group, EAMG group, DC treatment group, pcDNA3-TGF-β1-DCtreatment group, pcDNA3-DC control group and normal saline group. The rats were immunized with the AChR protein extracted from electric organ of Narcine timilei and CFA in the groups except normal group. 2×106 pcDNA3-TGF-β1-DCs/rat were injected subcutaneously into the backs of the rats which had been immunized 5 d earlier with AChR+CFA. The rats in DC treatment group, pcDNA3-DC control group and normal saline group were injected in parallel with untreated DC, pcDNA3-DC and normal saline, respectively. Seven weeks after the first immunization, the expressions of CD28 mRNA and CTLA-4 mRNA were detected by reverse transcription-polymerase chain reaction (RT-PCR), and the levels of B7-1 and B7-2 on the surface of PBMC were examined using flow cytometry. Results (1)The low expression of CD28 mRNA and rare expression of CTLA-4 mRNA were found in the normal rats, and both expressions increased markedly in EAMG rats (P＜0.001). Compared to those in EAMG group, the expression of CD28 mRNA decreased and CTLA-4 mRNA was upregulated after the treatment with pcDNA3-TGF-β1-DC (P＜0.05). There was no significant difference in the expressions of CD28 mRNA and CTLA-4 mRNA among the EAMG group, DC treatment group, pcDNA3-DC control group and normal saline group (P＞0.05). (2) The expressions of CD28, CTLA-4, B7-1 and B7-2 on the surface of PBMC were rare in normal rats, which increased significantly in EAMG rats (P＜0.001). The levels of CD28, B7-1 and B7-2 in pcDNA3-TGF-β1-DC group were lower than those in EAMG group (P＜0.01), but the level of CTLA-4 was higher than that in EAMG group (P＜0.05). They showed no statistically difference among the EAMG group, DC treatment group, pcDNA3-DC control group and normal saline group (P＞0.05). Conclusions The expressions of CD28/CTLA-4:B7 costimulatory molecules are abnormal in the rats with EAMG. The regulation of CD28/CTLA-4:B7 costimulatory pathways may play a critical role in the mechanism of the treatment with DC transfected with pcDNA3-TGF-β1 in the incipient EAMG rats.

OBJECTIVETo provide an cell model of immortalized lymphoblstoid B-cell lines for studying the biological characteristics of full-length hepatitis B virus (HBV) genome carrying the hot-spot mutations V60, G87, and L97.

METHODSV60, G87, and L97 mutation points were introduced into HBV p3.8 II plasmid containing 1.2 copy of HBV genome by means of site-directed mutagenesis. The HBV genome was amplified by PCR from p3.8 II and p3.8 II-V60, G87, L97 plasmid, and the PCR product was inserted into EBO-plpp eukaryotic expression vector. The recombinant vectors and the EBO-plpp vector were transfected into immortalized human lymphoblasts with lipofectamine 2000 and selected with hygromycin. Steady expression of the target genes was determined by RT-PCR, Western blotting and microparticle enzyme immunoassay.

RESULTSDNA sequence analysis indicated that the desired mutation was introduced into wild-type HBV DNA. HBsAg, HBeAg and HBcAg could be detected in EBO-HBV-transfected cell lysate or culture supernatant.

CONCLUSIONTransfectants that stably express HBV mutant antigen may provide a cell model to study the biological characteristics of HBV carrying hot-spot mutation in vitro.

B-Lymphocytes ; cytology ; virology ; Base Sequence ; Blotting, Western ; Cell Line, Transformed ; Cell Transformation, Viral ; DNA, Viral ; genetics ; Eukaryotic Cells ; metabolism ; Gene Expression Regulation, Viral ; Genetic Vectors ; Genome, Viral ; genetics ; Hepatitis B Core Antigens ; genetics ; metabolism ; Hepatitis B virus ; genetics ; metabolism ; Humans ; Molecular Sequence Data ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo study the effects of genotypes of HBV and HBeAg on the response to PEG-interferon alpha (PEG-IFN) in chronic hepatitis B (CHB) patients.

METHODSPCR-RFLP and S gene sequencing were conducted in 42 CHB patients.

RESULTSThe sustained response (SR) rates were 66.7% in genotype B and 27.3% in genotype C group. The P value was 0.039 by the Pearson Chi-square test, while it was 0.06 by the Fisher's exact test. The results suggested a trend that patients with genotype B HBV compared to genotype C had better SR to PEG-IFN therapy, although the difference was not significant. Results also showed that SR rate in patients with HBeAg-negative CHB (7/8 87.5%) was significantly higher than that in HBe+ CHB patients (8/21 38.1%, P < 0.05).

CONCLUSIONOur results indicate that HBV genotype and HBeAg, especially the later, are main factors for predicting PEG-IFN therapy response in CHB patients.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Genotype ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; Treatment Outcome

OBJECTIVETo study the immunologic function of dendritic cells (DCs) cultured in two kinds of hepatoma cell line's supernatant and the enhancing effects of carboxymethylpachymaran (CMP) on DCs.

METHODSDCs were harvested after stimulation by granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4 from umbilical cord blood using density-gradient centrifugation method. Cultured supernatant of two hepatoma cell lines (HepG2 and HepG2.2.15) were collected for condition medium (CM) according to a volume ratio of supernatant to incomplete RPMI-1640 medium, which was 3:1. CMP was dissolved in incomplete RPMI-1640 medium. Experimental groups were divided according to the culture medium, either CM or with CMP in it. DCs subsets CD83, CD86, CD1a, and d-related human leukocyte antigens (HLA-DR) were analyzed by flow cytometry. The proliferation ability of allogeneic T cells in mixed lymphocyte reaction (MLR) stimulated by DCs was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. IL-12p70, interferon-γ (IFN-γ), and nuclear factor κB (NF-κB) were detected by enzyme-linked immunosorbent assay analysis.

RESULTSThe proliferation of lymphocytes and secreting level of IL-12 and expression of phenotype of DCs cultured in two kinds of CM were lower than those of normal group (P <0.01). Compared with the normal group, groups treated with CMP showed a higher expression level of DCs subsets, lymphocyte reproductive activity, as well as IL-12 and IFN-γ secretion levels. Groups treated with CMP also demonstrated higher levels of DCs phenotype expression and IL-12 and IFN-γ secretion in supernatant of MLR and higher lymphocyte reproductive activity compared with CM group (P <0.05). Compared with the normal group, the expression level of NF-κB in DCs nuclear was higher in CMP groups but lower in two CM groups (P <0.05). After CMP was added, the NF-κB expression levels of two CM groups were increased compared with levels before CMP was added (P <0.05). However, there was no significant difference between the two CM groups (P >0.05).

CONCLUSIONSTwo kinds of hepatoma cell line's supernatant can inhibit the immunologic function of DCs. This suppressive effect may be related to the inhibition of NF-κB/Rel pathway. CMP may up-regulate the DCs function by activating the NF-κB/Rel pathway.

Carcinoma, Hepatocellular ; pathology ; ultrastructure ; Cell Line, Tumor ; Cell Shape ; Dendritic Cells ; drug effects ; immunology ; Glucans ; pharmacology ; Humans ; Immunophenotyping ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Liver Neoplasms ; pathology ; ultrastructure ; Lymphocyte Culture Test, Mixed ; Signal Transduction ; drug effects ; Subcellular Fractions ; drug effects ; Transcription Factor RelA ; metabolism

OBJECTIVETo study the prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi.

METHODSA total of 7407 people with Zhuang nationality, aged 16 and over were surveyed. Questionnaire was administered to each subject under survey, while X-ray and serum rheumatoid factor were carried out to all those who gave positive answers. Physical examinations were done to the suspicious cases by experts on rheumatism.

RESULTSwere compared with those in 6826 people of Han ethnicity living in the same district.

RESULTSThe prevalence rates of rheumatoid arthritis in Zhuang nationality population was 0.27% when comparing to the Han population which was 0.28% (P > 0.05). Rates of awareness on rheumatoid arthritis in Zhuang and Han population were 5.0% and 10.5% (P > 0.05). After the diagnosis of rheumatoid arthritis was made and among patients who had received the treatment, the rates were 0% vs. 5.25%.

CONCLUSIONThe prevalence rate of rheumatoid arthritis in Zhuang nationality population of Nanning, Guangxi was not significantly different than that in Han ethnic group. However, the rates on awareness and the treatment of rheumatoid arthritis were still under satisfaction.

Adult ; Aged ; Arthritis, Rheumatoid ; epidemiology ; Asian Continental Ancestry Group ; ethnology ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Prevalence

OBJECTIVETo obtain the coronary artery and coronary sinus (CS) and its tributaries imaging with multislice computed tomography (MSCT), measure the distance between coronary artery and CS and its tributaries and analyze their spatial relationships.

METHODSThe MSCT scans of 117 patients (67 men, 50 women, age 56 +/- 10 years) were obtained, 3D image reconstructed and the vessels courses evaluated. The concomitant distances and spatial relationships of the vessels were determined.

RESULTSRight coronary artery domination was found in 107 cases (91.4%), left coronary artery domination in 7 cases (6.0%), and co-domination in 3 cases (2.6%). Left circumflex artery (LCX) was concomitant with CS or the great cardiac vein (GCV) in 81 cases (69.2%), intersected with left posterior vein in 62 cases (53.0%) and with middle cardiac vein (MCV) in 5 cases (4.3%), respectively. The dominant coronary artery branched out into the posterior descending artery (PDA) and the left posterior artery (LPA) in 112 cases (95.7%). PDA was concomitant with MCV in 93 cases (79.5%) and intersected with MCV in 44 cases (37.6%). LPA was intersected with MCV in 106 cases (90.6%), and concomitant with CS in 50 cases (42.7%).

CONCLUSIONSMSCT is a reliable tool to visualize the relationship between coronary artery and CS and its tributaries. Owing to the multiple possibilities inherent to this technique, MSCT has broad potential for more clinical use.

Adult ; Aged ; Aged, 80 and over ; Coronary Angiography ; Coronary Sinus ; anatomy & histology ; diagnostic imaging ; Coronary Vessels ; anatomy & histology ; Female ; Humans ; Male ; Middle Aged ; Tomography, Spiral Computed

OBJECTIVETo evaluate the efficacy and safety of nateglinide, a new antidiabetic agent, in the treatment of type 2 diabetes.

METHODSA total of 219 treatment-naïve patients with type 2 diabetes from 6 centers were enrolled in this study and blindly divided into nateglinide group (n = 105) and repaglinide group (n = 114). In all patients, the disease was confirmed for at least three months. The whole observation lasted for 12 weeks. The efficacy indicators measured include glycohemoglobin A1c (HbA1c), fasting blood glucose, and 2 hours postprandial blood glucose, and the safety parameters measured included renal and hepatic function, serum lipids, and blood and urea profiles.

RESULTSSimilar decreases in fasting blood glucose, 2 hours postprandial blood glucose, and HbA1 c were found in both nateglinide group and repaglinide group without significant differences. No severe adverse events were noted. The hypoglycemia event reports were not significantly different between these two groups.

CONCLUSIONNateglinide is an effective and safe drug in treating type 2 diabetes.

Blood Glucose ; drug effects ; Cyclohexanes ; administration & dosage ; adverse effects ; therapeutic use ; Diabetes Mellitus, Type 2 ; drug therapy ; Drug Administration Schedule ; Female ; Humans ; Hypoglycemic Agents ; administration & dosage ; adverse effects ; therapeutic use ; Male ; Middle Aged ; Phenylalanine ; administration & dosage ; adverse effects ; analogs & derivatives ; therapeutic use ; Treatment Outcome

Objective To develop a detergent for decontamination of Co2+ and Mn2+ on skin.Methods Single-factor experimental and orthogonal experimental designs were performed to study the formula composition of the decontaminant.The skin irritation experiment was performed and assessed according to the standard method.The detergent was prepared according the conventional process of showering gel.The pH,ethylenediaminetetraacetic acid (EDTA) level,total active substances of the detergent,and its stability were evaluated according to the chemical method recommended in the national standard GB/T 13173-2008.The decontamination efficiency on stable isotopes of Co2+ and Mn2+ contamination was measured on the back of hand skin of volunteers.Results The formula composition of the decontaminant was obtained through the orthogonal experiment.The pH value of the detergent was 6.99,total active substance was 20.49％ and the content of EDTA was 5.99％.After being kept at-5 ℃ and 40℃℃ for 24h,the decontaminant showed no strange smell,no precipitation,no discoloration and still kept transparent.The decontamination effects on Co2+ and Mn2+ contaminated on hand skin were 103.13％ ± 0.05％ and 100.62％ ± 0.09％,respectively,which was significantly higher than that of distilled water (81.77％ ± 0.23％ and 79.63％ ± 0.23％,P＜0.01,respectively).Conclusion The decontaminant has a high effect on decontamination of Co2+ and Mn2+ polluted on skin,and is hopeful to be developed as an effective detergent on radioactive isotopes contamination.