In order to solve the difficucties of renaturation and immunogenicity of new bifunctional fusion protein GAD,inclusion bodies of GAD were modified by PEG-maleimide.Conformational changes of the modified GAD were compared by circular dichroism and tryptophan fluorescence spectroscopy.The biological activity was verified by oral glucose tolerance test and lipid scavenging.The results showed that PEG-maleimide completed the specific-point modification of GAD,and improved its refolding efficiency.The secondary and tertiary structures of mPEGylated GAD were consistent with that of GAD.PEG-GAD has significant hypoglycemic and lipid-lowering effects (P ＜0.001) with longer half life in vivo and lower immunogenicity (P ＜0.01).This study provides effective strategies for the development of strongly hydrophobic peptide drugs.

Objective To investigate the role of chemokine ligand 21 (CCL21) in the spinal cord in tibia bone cancer pain (BCP) in rats.Methods Forty adult female SD rats weighing 160-180 g were randomly divided into 5 groups (n=8 each):sham operation group (group Ⅰ ); sham operation + CCL21 neutralizing antibody group (groupⅡ); BCP group (group [); BCP + PBS group (group Ⅳ); BCP + control IgG group (groupⅤ)and BCP + CCL21 neutralizing antibody group (group Ⅵ).BCP was induced by inoculating Walker-256 mammary gland carcinoma cells into the rat tibia medullary cavity in groups Ⅲ-Ⅵ.PBS 15 μl,IgG 10 μg and CCL21 neutralizing antibody 10 μg were injected intrathecally (IT) at 14 days after intra-tibial injection of Walker-256 mammary gland cancer cells in groups Ⅳ- Ⅵ respectively.Mechanical withdrawal threshold to yon Frey filament stimulation (MWT) was measured at 1 day before (To,baseline) ; 7 and 14 d after Walker-256 cell injection (T1,T2)and at 0.5,1,2,4,8,12,24 and 48 h after intrathecal injection (T3-10 ).Results Intra-tibial injection of Walker-256 mammary gland cancer cells significantly decreased MWT as compared with the baseline values in administration of CCL21 neutralizing antibody at T5-8 as compared with MWT before intrathecal administration at T2 in group Ⅵ.MWT was significantly lower in groups Ⅲ- Ⅳ than in groups Ⅰ and Ⅱ.MWT was significantly higher at T5-8 in group Ⅵ than in groups Ⅲ - Ⅴ.Conclu]sion CCL21 in the spinal cord is involved in the maintenance of tibia BCP in rats.

Acute Disease ; Child ; Cytidine Deaminase ; genetics ; Humans ; Leukemia ; genetics ; Polymorphism, Single Nucleotide ; genetics

Objective To investigate effects of intrathecal methotrexate on mechanical allodynia in rats with tibial bone cancer pain.Methods Forty-eight female SD rats weighing 150-180 g were randomly divided into 6 groups ( n =8 each):group Ⅰ sham operation + artificial cerebrospinal fluid(SA group),group Ⅱ sham operation + methotrexate 200 μg(SM group),group Ⅲ bone cancer pain + artificial cerebrospinal fluid(CA group),group Ⅳ-Ⅵ bone cancer pain + different doses of methotrexate (CM1-3 groups).The model of tibial bone cancer pain was induced by injecting Walker-256 cell into the tibial marrow cavity.CA and CM1-3 groups were intrathecal injected artificial cerebrospinal fluid,methotrexate 50,100 and 200 μg.SA and SM200 groups were intrathecal injected artificial cerebrospinal fluid and methotrexate 200 μg.The mechanical withdrawl threshold (MWT) was measured at day 1 before Walker-256 injection (baseline),7 day after injection (T0 ) and 2,4,8,24 hour and 1,3,5,7 days after intrathecal injection ( T1-8 ).Results Compered with the baseline,MWT was decrease in CA and CM1-s groups.Competed with To,MWT was decreased at T5-8 in CA group,MWT was increased at T3-5 in CM1 group,at T2-6 in CM2 group and at T2-7 in CM3 groups.MWT was decrease in CA and CM1-3 groups as compered with SA group; MWT was increased at T4-7 in CM1 group and at T3-7 in CM2 and CM3 groups.Conclusion Intrathecal injection of methotrexate can reduce tibial bone cancer pain in rats.

Dengue virus (DENV) is a re-emerging disease transmitted by the Aedes mosquitoes and has become a major public health problem in southern China. Currently, no antiviral drug or effective vaccine exist to control this disease. The chimeric DENV structural protein vaccine cannot elicit balanced levels of protective immunity to each of the four viral serotypes; therefore, non-structural protein components may be required to construct an effective DENV vaccine. The Dengue virus non-structural 1 (DENV NS1) protein plays a critical role in viral pathogenesis and protective immunity. Therefore, immunity to Dengue 1-4 NS1 subtypes may be crucial for the prevention of severe disease. This review attempts to provide an overview about the structure and function of DENV NS1.
Animals ; Dengue ; immunology ; prevention & control ; virology ; Dengue Vaccines ; chemistry ; genetics ; immunology ; Dengue Virus ; chemistry ; genetics ; immunology ; Humans ; Viral Nonstructural Proteins ; chemistry ; genetics ; immunology

Objective To make a Meta-analysis of the effectiveness and safety of Shenling Baizhu Powder (SBP) for the treatment of ulcerative colitis(UC),thus to provide evidence for the clinical treatment of ulcerative colitis.Methods Randomized controlled trials (RCTs) of SBP combined with western medicine vs western medicine in treating ulcerative colitis were included.The quality of RCTs was assessed by the Cochrane scale.A Meta-analysis was performed for the clinical efficacy,improvement of disease activity index (DAI) levels of the included trials.Results A total of 19 RCTs were included,involving 1498 cases.The results of Metaanalysis showed that compared with the western medicine group,the combined risk ratio(RR) of clinical efficacy in the SBP combined group was 1.55,95％ confidence interval (CI) being (1.39,1.72).The subgroup analysis based on control medicine showed that the combined RR of combined use of sulfasalazine or mesalazine/Olsalazine was 1.46,1.59 [95％CI (1.19,1.77) vs 95％CI (1.40,1.80)].The subgroup analysis based on different treatment courses showed that the combined RR of 1-30 days,31-60 days,61-90 days was 1.42,1.69,1.47 [95％CI (1.18,1.70) vs 95％CI (1.44,1.97) vs 95％CI (1.15,1.88)] respectively.The differences were significant (P ＜ 0.05).(2) The differences of the two groups on the improvement of DAI and inflammatory factors levels of interlekin-17(IL-17),IL-23,tumor necrosis factor alpha(TNF-α),C-reactive protein(CRP) were statistically significant(P ＜ 0.05).(3)The sensitivity analysis of the primary outcomes showed a higher homogeneity in the literatures and the funnel plot analysis showed no evidence of publication bias.Conclusion Compared with western medicine,SBP combined with western medicine has better clinical efficacy for the treatment of UC,and the combined use of mesalazine/Olsalazine medicated for 30-60 days is more effective on improving DAI and inflammatory factors levels.However,for the low quality of the included literatures and insufficient experimental design,the conclusion needs more evidence from large sample-size randomized double-blind controlled trials.

Objective To examine the inter- rater reliablity of Myoton-3 Myometer in assessment of muscle tone in healthy subjects.Methods 20 healthy volunteers were assessed their muscle tone of biceps brachii and flexor carpi radialis muscles in relaxed state with Myoton-3 by 2 testers within 24 h. The frequency of damping oscillations (F value) measured by Myoton-3 was as the characteristic of the muscletone. The intraclass correlation coefficient (ICC) and Bland-Altman analysis were performed. Results The ICC was 0.72~0.88 and 0.79~0.89 in triple scan and ten-time scan pattern, respectively. The Bland-Altman analysis revealed no systematic errors between testers. ConclusionThe Myoton-3 Myometer is reliable between testers for measuring the muscle tone in healthy adults.

OBJECTIVE: To explore the conversion of resin monomer, the change of inorganic component and the influencing factors on the oxygen inhibition layer formed on the cured surface of resin cement. METHODS: Three kinds of resin cement were divided into three groups: (1) light-cured group: RelyX Veneer, NX3 (light-cured), Variolink N; (2) dual-cured group: RelyX U200 Automix, NX3 (dual-cured), Multilink Speed; (3) chemically-cured group, and the above 3 types of dual-cured resin cement cured without illumination could be used as chemically-cured resin cement. Each sample was provided with and without oxygen exposure of two matching surfaces, cured respectively, and the variables of light intensity and illumination time were set in the light-cured group and the dual-cured group. Scanning electron microscopy was used to observe the samples' surface morphology. Energy dispersive spectrometer was used to analyze the samples' composition of surface elements. Confocal Raman spectroscopy was used to measure the monomer conversion of resin cement and to obtain the thickness of the oxygen inhibition layer. RESULTS: (1) On the surface of cured resin cement, the weight percentage of oxygen element in the aerobic side was higher than that in the anaerobic side (P < 0.05), and the weight percentage of inorganic element was lower than that in the anaerobic side (P < 0.05). (2) The surface monomer conversion of the cured resin cement on the aerobic surface was significantly lower than that on the anaerobic surface (P < 0.05), and the surface monomer conversion on the aerobic surface and the anaerobic surface was the lowest in the chemically-cured group (P < 0.05), the dual-cured group was the highest (P < 0.05), and the light-cured group was between them. With the increase of light intensity or illumination time, the surface monomer conversion increased (P < 0.05). (3) The thickness of the oxygen inhibition layer was the thickest in the chemically-cured group [(40.27±2.81) μm](P < 0.05), the thinnest in the dual-cured group [(21.87±5.42) μm](P < 0.05) and light-cured group [(23.73±3.84) μm] was between them. With the increase of light intensity or illumination time, the thickness of the oxygen inhibition layer of resin cement decreased (P < 0.05). CONCLUSION When resin cement is exposed to oxygen, it will form an oxygen inhibition layer, its surface's inorganic filler is less, the surface monomer conversion is lower. The surface monomer conversion and the thickness of oxygen inhibition layer are affected by curing mode and illumination factors.
Materials Testing ; Oxygen ; Resin Cements

Objective To investigate the role of chemokine ligand 2 (CCL2) in the spinal cord expression in a rat model of tibia bone cancer pain.Methods Eighty-four female SD rats weighing 160-180 g were randomly divided into 3 groups ( n =28):control group (group C),sham operation group (group S) and tibia bone cancer pain group (group P).Tibia bone cancer pain was induced by intra-tibial inoculation of Walker-256 breast cancer cells.Paw withdral threshold to mechanical stimulation (MWT) was measured with von Frey filaments at 1 d before and at 1,3,7,10,14 and 21 d after inoculation.Six rats in each group were sacrificed after the measurement of MWT at 1 d before inoculation and at 7,14 and 21 d after inoculation.Lumbar 4-6 segments of the spinal cord were removed for determination of the expression of CCL2 by ELISA.The coexpression of CCL2 with Iba-1 (a specific marker of microglia),GFAP(a specific marker of astrocyte) and NeuN (a specific marker of neuron) was determined by double immunofluorescence assay after the measurement of MWT at 14 d after inoculation in group P.Results Compared with groups C and S,MWT was significantly decreased from 7 d to 21 d after inoculation,the expressive of CCI-2 in the spinal cord up-regulated at 7,14 and 21 d after inoculation in group P ( P ＜ 0.05).CCL2 was expressed in the microglia and astrocyte but not in neuron in the spinal cord dorsal horn in a rat model of tibia bone cancer pain.Conclusion Release of CCL2 from microglia and astrocytes in the spinal cord was involved in mechanical hyperalgesia in a rat model of tibia bone cancer pain.

Objective A comparative proteomic method was utilized to analyze serum proteins among pancreatic cancer patients,pancreatic benign tumor group,chronic pancreatitis group and normal control group to discover a new potential specific early diagnostic marker.Methods Comparative analysis on the pancreatic peripheral blood protein profiling from 40 pancreatic cancer patients,10 benign tumor patients,10 chronic pancreatitis patients and 40 cancer-free controls from May 2009 to April 2011 was carried out by 2D differential gel electrophoresis (2D-DIGE) and differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS).Results Three differentially expressed proteins,Hemopexin (Hpx),Ficolin 3 (FCN3) and Serum amyloid P-component (SAP) was identified.Those proteins were higher expression in pancreatic cancer group compared with benign tumor group,chronic pancreatitis group and normal control group.Each point in pancreatic cancer expression were 1.57,1.99,1.63 times than normal control expression,respectively (P ＜0.05).Conclusions In this study,the identified proteins,Hpx,FCN3 and SAP may be as potential specific early diagnostic markers of pancreatic carcinoma.2D-DIGE and MALDI-TOF-MS technology in screening specific serum tumor markers of pancreatic cancer has a well repeatability and stability.