Objective To optimize the prescription and preparation technology of brucine nanostructured lipid carriers (B-NLC). Methods The method of "the solvent emulsification ultrasound" was used to prepare B-NLC. The prescription and preparation was optimized using a single factor method combined with central composite design-response surface methodology (CCD-RSM). Results The resultant B-NLC was transparent liquid with light blue opalescence. The optimal conditions were that the dosage of drugs was 1.28 mg, the mass concentration of poloxamer 188 was 1.08%, and the ratio of solid lipid to liquid lipid was 1.45:1. The obtained NLC showed the average particle size of (136.89 ± 4.23) nm with a polydispersity index of 0.289 ± 0.005 and a zeta potential of (-34.46 ± 0.31) mV. The entrapment efficiency was calculated to be (68.98 ± 2.06)%, and the drug loading content was (1.90 ± 0.06)%. Conclusion B-NLC prepared by solvent emulsification ultrasound had a high entrapment efficiency and a narrow particle size distribution. The method was easy and simple and can be used to optimize the prescription and preparation of B-NLC, which provides a foundation for the further in vivo research of brucine.

Background: The side effects of glucocorticoid in treatment of systemic lupus erythematosus (SLE) have been the focus of debate, and our preliminary study indicates that ginsenosides can enhance the efficacy of dexamethasone. Objective: To observe the effects of ginsenosides combined with prednisone in SLE patients. Design, setting, participants and interventions: A total of 60 SLE patients from Department of Rheumatology and Immunology, Changhai Hospital, Second Military Medical University, were randomly divided into treatment group and control group, with 30 patients in each group. Patients in the treatment group were given routine treatment with prednisone plus ginsenosides, while those in the control group were given routine treatment with prednisone plus placebo. They were all treated for 3 months. Main outcome measures: After three-month treatment, syndrome score in traditional Chinese medicine (TCM), total response rate and symptom improvement rate were measured and evaluated. Results: Twenty-eight cases in treatment group and twenty-seven cases in control group were included in analysis. The total response rates in the treatment group and control group were 89.28% and 66.67% respectively, and there was a significant difference between the two groups (P<0.05). After treatment, the TCM syndrome scores in the two groups were lower than those before treatment (P<0.01), and prednisone plus ginsenosides was better in decreasing the TCM syndrome score than prednisone plus placebo (P<0.05). The symptoms were improved in the treatment group as compared with the control group (P<0.05). Conclusion: Prednisone combined with ginsenosides can increase the clinical effective rate and improve the clinical symptoms of SLE patients.

Objective To evaluate the histologic changes in the dermis and the changes of the rate of type Ⅲ and type Ⅰ collagen by the radiofrequency device. Methods The effects of radiofrequency current on the dermis were observed. Ten rabbits were treated by radiofrequency, and the histologic change in the dermis were observed by H-E staining and Sirius red staining. Results After RF treatment, the fibers in the dermis appeared more compact and the quantity of the type Ⅲ (red) and type Ⅰ (green) collagen were both increased. The fibers in the dermis appeared more compact and the rate of type Ⅲ and type Ⅰ collagen was increased. It was also found that a significant proliferation of dermal collagen was observed in 8 days after treatment. As time went by, the proliferation of dermal collagen was more pronounced, and the rate of type Ⅲ was increased. Conclusion The radiofrequency current can increase the quantity of collagen in the dermis and increase the rate of type Ⅲ and type Ⅰ collagen, which may be one of the key mechanisms of facial rejuvenation by RF.

Antibodies ; immunology ; therapeutic use ; Antigens, CD20 ; immunology ; Antilymphocyte Serum ; therapeutic use ; CD3 Complex ; immunology ; Diabetes Mellitus, Type 1 ; drug therapy ; immunology ; Humans ; Immunotherapy ; methods

AIMTo test the stability of marine polysaccharide drug sulfated polyguluronic acid ester.

METHODSFour methods including high performance gel chromatography (HPGC), poly-acrylamide gel electrophoresis (PAGE), UV scan of absorbance between 200 and 800 nm and gelatin nephelometry were established. Samples were tested in high temperature, high humidity, strong light and accelerated test conditions. The methods were used to test the changes of the parameters including molecular weight, molecular weight distribution, absorbance between 200 and 800 nm, free sulfate, with which we could estimate the stability of sulfated polyguluronic acid ester could be estimated.

RESULTSThe four methods were suitable to test the stability of sulfated polyguluronic acid ester and the sample were stable in the conditions as before except in high temperature.

CONCLUSIONSulfated polyguluronic acid ester has good stability.

Chromatography, Gel ; methods ; Drug Stability ; Electrophoresis, Polyacrylamide Gel ; methods ; Molecular Weight ; Polysaccharides, Bacterial ; chemistry ; Spectrophotometry, Ultraviolet ; methods ; Temperature

OBJECTIVETo explore the effects of bisphenol A (BPA) exposure on toxicity characteristic and OCT4 and SOX2 gene expression of mouse embryonic stem cells (mESC).

METHODSmESC were cultured, and treated with the doses of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L respectively of BPA and DMSO (the solvent control group)for 24 hours, and three groups of cells were treated with the same method. The morphological changes of mESC in the control and exposure groups were observed through an inverted microscope. Cell counting kit 8 (CCK8) was used to detect the effects of BPA on proliferation of mESC, and based on the results, the half inhibitory concentration (IC50) was calculated. Real-time fluorescent quantitative polymerase chain reaction (RT-QPCR) and western blotting were used to detect the expression of OCT4 and SOX2.

RESULTSBPA had certain toxicity on mESC, the treatment of BPA significantly increased cell toxicity in a concentration-dependent manner, and the IC50 was 4.3×10(-4) mol/L, combined with the BPA exposure concentration of the environment and the related literature, eventually taking the five concentrations of 10(-8), 10(-7), 10(-6), 10(-5), 10(-4) mol/L as the experimental groups. The mESC morphology were effected after the treatment of BPA for 24 h, compared with the control group, the number of cells decreased, appearing some floating cells, and the cell cloning became irregular and differentiation in the higher concentration groups. The OCT4 mRNA expression level in the 10(-7) mol/L (1.146 ± 0.087), 10(-6) mol/L (1.156 ± 0.030), 10(-5) mol/L (1.158 ± 0.103) and the 10(-4) mol/L (1.374 ± 0.053) dose group were all significantly higher than the control group (1.000 ± 0.000) (t values were -2.384, -2.953, -3.203, -4.021 respectively, P value all < 0.05). Meanwhile, the SOX2 mRNA expression level in the 10(-4) mol/L (1.113 ± 0.052) were higher than the control group (1.000 ± 0.000) (t value was -2.765, P value < 0.05). Moreover, the OCT4 protein expression level in the 10(-5) mol/L (1.360 ± 0.168) and 10(-4) mol/L (1.602 ± 0.151) were all significantly higher than the control group (1.000 ± 0.000) (t values were -3.538, -4.002 respectively, P value all < 0.05), while no obvious change of the SOX2 protein expression level was detected in all treated groups.

CONCLUSIONBPA in a certain dose range could upregulate the expression of OCT4 gene in mouse embryonic stem cells while had no significant effect on the expression of SOX2 gene.

Animals ; Benzhydryl Compounds ; toxicity ; Cells, Cultured ; Embryonic Stem Cells ; drug effects ; metabolism ; Gene Expression ; Mice ; Octamer Transcription Factor-3 ; genetics ; Phenols ; toxicity ; SOXB1 Transcription Factors ; genetics ; Signal Transduction ; drug effects

ObjectiveTo evaluate the adverse events occurred during tumour necrosis factor (TNF)-αblocker treatment in Chinese Han population patients with ankylosing spondylitis (AS).MethodsThis study had enrolled 369 Chinese Han population patients with ankylosing spondylitis.They all received TNF-αblocker treatment in the hospital.All 1011 administration were recorded in total.All of them were evaluated for adverse events 2 hours after injection,126 of them had received long-term TNF-α blocker injection,and they were followed-up at week 8,12,52,104.Mild immediate adverse events and long-term adverse events were all counted.SPSS 10.0 software package was used for Fisher's exact test.ResultsThree hundred and sixty-nine patients had 1011 administrations in total,652 had received rhTNFR:Fc,316 had infliximab,21had etanercept,22 had adalimumab injections.Adverse events 2 hours after injection were:17 (2.6％) for rhTNFR:Fc,12 (3.8％) for infliximab,0 for etanercept,1 (4.5％) for adalimumab.Twenty adverse events were mild(12 for rhTNFR:Fc,9 for infliximab),5 events were moderate(3 for rhTNFR:Fc,1 for infliximab,1 for adalimumab),4 events were severe(2 for rhTNFR:Fc,2 for infliximab).The frequency of adverse events were comparable between rhTNFR:Fc and Infliximab injection in immediate adverse reactions (P=0.31).One hundred and twenty-six (69 rhTNFR:Fc,57 infliximab) patients had long-term usage,and were followed-up at week 8,12,52,104,39 patients had adverse reactions:20 (51.3％) for rhTNFR:Fc,19(48.7％) for infliximab.Thirty-seven patients had infectious events(94.9％ ),1 neurological event(2.6％),and 1 patient had tuberculosis relapse (2.6％).Outcomes were comparable with rhTNFR:Fc and infliximab in long-term usage(P=0.69).ConclusionAttention should be paid to the above events in Chinese Han patients with ankylosing spondylitis who were treated with TNF-α blocker treatment.Special attention should be paid to those patients who are in their third or fourth injection.The occurrence of immediate reaction or long-term adverse events between rhTNFR:Fc and infliximab are comparable.

OBJECTIVETo investigate the perfusion restoration of type 1 diabetic mouse under the setting of surgically induced hind limb ischemia and the number and function of bone marrow endothelial progenitor cells (EPCs).

METHODSForty mice were randomly divided into two groups: one group was injected with alloxan through tail vein to induce type 1 diabetes mellitus, and another group was set as control group. All mice were surgically induced to hind limb ischemia. Blood flow was monitored with Laser Doppler perfusion imaging for 4 weeks after artery ligation. Ten mice in each group were sacrificed and muscle tissues were harvested for histological detection. The remaining mice were sacrificed 7 days after surgery, bone marrow mesenchymal stem cells were harvested and EPCs were analyzed by flow cytometry and then were collected to culture for functional detection.

RESULTSAll mice received alloxan injection showed typical symptoms of type 1 diabetes mellitus. Restoration of blood flow was significantly slower in type 1 diabetic mice with lower level of vascular density in ischemic muscles than control group (P < 0.001, P < 0.05). The number and function of EPCs (CD34 and vascular endothelial growth factor receptor 2 double positive cells) in type 1 diabetic mice were significantly lower than that in control mice (P < 0.05).

CONCLUSIONSThe spontaneous angiogenesis is attenuated with a decreased number and function of EPCs in the setting of type 1 diabetes mellitus. This may partly explain why diabetic patients with peripheral artery diseases have more aggressive disease and poorer outcome.

Animals ; Antigens, CD34 ; analysis ; Cell Count ; Diabetes Mellitus, Experimental ; complications ; pathology ; Diabetes Mellitus, Type 1 ; complications ; Hindlimb ; blood supply ; Ischemia ; complications ; physiopathology ; Mesenchymal Stromal Cells ; chemistry ; Mice ; Reperfusion ; Vascular Endothelial Growth Factor Receptor-2 ; analysis

We separated proteins of the middle silk gland through high resolution two-dimensional polyacrylamide gel electrophesis, and identified the high expressional proteins using MALDI-TOF-MS and analyzed the PMF in protein database by GPMAW( General Protein/Mass Analysis for Windows) . The protein database was forecasted through silkworm genome database. More than 500 spots were obtained from each gel by silver stain and more than 100 protein spots were obtained from gel by Coomassie brilliant blue stain. Most of them were distributed in the area from 15kD to 90kD with pH 3.5-7. Among the 25 Coomassie brilliant blue stained spots identified by MALDI-TOF-MS, more than 60% have relatively strong signal. Comparing with the result of using Mascot, the method using PMF database of silkworm not only can identify some known proteins, but also can identify some unknown proteins that have been forecasted in silkworm genome database. Accordingly, we founded a complete set of method that fit for researching proteome of silkworm.
Amino Acid Sequence ; Animals ; Bombyx ; metabolism ; Databases, Protein ; Electrophoresis, Gel, Two-Dimensional ; Hydrogen-Ion Concentration ; Insect Proteins ; analysis ; chemistry ; Molecular Sequence Data ; Molecular Weight ; Proteome ; analysis ; chemistry ; Proteomics ; methods ; Silk ; metabolism ; Software ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVESTo explore the relationships between the peripheral blood levels of CD61, CD63, PAC-1 and the incidence of acute rejection and tubular necrosis after renal transplantation, and recovery of the graft function.

METHODSThe peripheral blood levels of CD61, CD63, and PAC-1 of 86 patients with uremia in different stages before and after transplantations were analyzed by flow cytometry. The patients were divided into three groups: (1) twenty-nine patients with normal grafts function, (2) hirty with acute rejection and (3) twenty-seven with acute tubular necrosis. The patients with acute rejection were randomly divided into treatment group with anticoagulants and cntrol group.

RESULTSThe peripheral blood levels of CD61, CD63 and PAC-1 significantly increased (P < 0.05) in the patients with acute rejection, in comparison with those with normal grafts function and those with acute tubular necrosis. The peripheral blood levels of CD61, CD63 and PAC-1 in patients with acute rejection in anticoagulants therapy was lower, recovery time of the grafts function was shorter, one-year survival rates of patients and grafts were higher, as compared with those of controls.

CONCLUSIONSThe patients with acute rejection have significantly high peripheral blood levels of CD61, CD63 and PAC-1 before transplantation, however, these values in patients with acute tubular necrosis are not high, this suggesting that acute rejection might relate to platelet activation, while acute tubular necrosis might not relate to it. After anticoagulants therapy in patients with acute rejection, the grafts function might recover faster and their one-year survival rates and grafts might be higher in those with CD61, CD63 and PAC-1 decreasing remarkably.

Adult ; Aged ; Antigens, CD ; blood ; Dual Specificity Phosphatase 2 ; Female ; Graft Rejection ; Humans ; Integrin beta3 ; blood ; Kidney ; physiopathology ; Kidney Transplantation ; Male ; Middle Aged ; Platelet Activation ; Platelet Membrane Glycoproteins ; Protein Phosphatase 2 ; Protein Tyrosine Phosphatases ; blood ; Tetraspanin 30