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Objective:To report thigh muscle magnetic resonance imaging (MRI)tests of four Chinese patients with dystrophinopathy with edema changes in adductor longus muscles that mimics adductor en-thesopathy.Methods:Four boys,who were from four unrelated families and aged from 5 to 11 years, were investigated because of the clinical manifestations including myalgia or muscle weakness or the inci-dental findings of elevated serum creatine kinase levels,and were diagnosed with dystrophinopathy by gene test of Duchenne muscular dystrophy (DMD).Their creatine kinase levels were increased from 4 087 IU /L to 32 700 IU /L (Normal range:75 -175 IU /L).The muscle biopsy of three patients all demonstrated a dystrophic pattern including necrosis,regeneration,hypertrophy,atrophy and connective tissue proliferation,with different proportions of dystrophin-negative muscle fibers.The gene test of DMD showed an out-frame deletion of exons in three of the four patients,involving either exons 45 or exons 49 -52 deletion or exon 62 duplication,and c.2665 C >T with nonsense mutation in the other one. Muscle MRI tests of the bilateral thighs were performed with T1 weighed sequence and slow tau inversion recovery sequence.The degree of fatty infiltration changes was scored.Results:MRI of the thigh mus-cles showed mild to severe fatty infiltration changes in T1 weighed sequence with the total scores from 2 to 13.The most severe fatty infiltration changes were in the long head of biceps femoris and adductor mag-nus.Obvious hyperintensities appeared mainly in the adductor longus muscles on slow tau inversion re-covery (STIR)images in all the patients without any abnormal signals in the attachment of the ligament, indicating edema changes of the adductor longus muscles which mimiced adductor enthesopathy.Two of the four patients presented with edema changes in the bilateral adductor longus muscles,while the other two were with only unilateral changes.Furthermore,other thigh muscles,including adductor magnus, semitendinosus,sartorius and rectus femoris muscles,could also have mild edema changes in two of the four patients.Conclusion:Dystrophinopathy can manifest as edema changes in the adductor longus mus-cles in thigh muscle MRI tests,which is a typical lesion in adductor enthesopathy.The adductor longus muscles in the dystrophinopathy patients may be easy to be impaired due to traction injury during sports.
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Objective:To describe a method based on analysis of the histogram of intensity values pro-duced from the magnetic resonance imaging (MRI)for quantifying the degree of fatty infiltration. Methods:The study included 25 patients with dystrophinopathy.All the subjects underwent muscle MRI test at thigh level.The histogram Mvalues of 250 muscles adjusted for subcutaneous fat,representing the degree of fatty infiltration,were compared with the expert visual reading using the modified Mercuri scale.Results:There was a significant positive correlation between the histogram Mvalues and the scores of visual reading (r =0.854,P <0.001).The distinct pattern of muscle involvement detected in the pa-tients with dystrophinopathy in our study of histogram M values was similar to that of visual reading and results in literature.The histogram M values had stronger correlations with the clinical data than the scores of visual reading as follows:the correlations with age (r =0.730,P <0.001 )and (r =0.753, P <0.001);with strength of knee extensor (r =-0.468,P =0.024)and (r =-0.460,P =0.027) respectively.Meanwhile,the histogram Mvalues analysis had better repeatability than visual reading with the interclass correlation coefficient was 0.998 (95% CI:0.997 -0.998,P <0.001)and 0.958 (95%CI:0.946 -0.967,P <0.001)respectively.Conclusion:Histogram Mvalues analysis of MRI with the advantages of repeatability and objectivity can be used to evaluate the degree of muscle fatty infiltration.
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Objective To analyze the characteristics of fat infiltration into the muscles of patients with Becker and Duchenne muscular dystrophy (DMD) so as to provide a guide for rehabilitation therapy.Methods Twenty-three children with Becker muscular dystrophy (BMD) and 47 with DMD who had never been treated with glucocorticoids were enrolled.MRI was performed on both of their thigh muscles.T1 weighted images were used to assess the fat infiltration of their thigh muscles using a 0-5 modified version of Mercuri's scale.The progression of fatty infiltration of the thigh muscles in BMD was analyzed using descriptive statistics.The differences in fat infiltration between BMD and DMD were analyzed using rank sum tests.Results In patients with BMD the adductor magnus most often showed severe fat infiltration,followed by the biceps femoris,quadriceps,semimembranosus and semitendinosus,while the sartorius,gracilis and adductor longus had the lowest percentages of severe fat infiltration.Among the BMD patients the adductor magnus,biceps femoris and quadriceps showed moderate to severe involvement at the age of 8 to 9.The semimembranosus and semitendinosus showed moderate to severe involvement at the age of 10 to 11,and the sartorius,gracilis and adductor longus showed mild to moderate involvement after 15 years of age.Among the age groups of 8,9,10 and 11 years old,the median total fat infiltration scores were 10,22,28 and 25 respectively among the BMD patients,and 29,34,34 and 30 respectively among the DMD patients.At age 8 significant differences between the BMD and DMD patients were observed in the infiltration scores of the adductor magnus,biceps femoris,vastus lateralis,rectus femoris,vastus medialis,vastus intermedius and in the total scores.At age 9 significant differences persisted in the scores of the adductor magnus,rectus femoris,vastus medialis,vastus intermedius and the total scores.Conclusions The muscle MRIs showed significant differences in the degree of fatty infiltration between BMD and DMD patients.These findings may be useful when designing therapeutic regimens and rehabilitation programs for patients with BMD and DMD.
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Objective:To study the effects of Anti-DR5 mAb on inducing synovial cells of adjuvant arthritis (AA) rats and its mechanisms.Methods:AA was induced by CFA in rats.The synovial cells of rats were separated and cultured in vitro system.The apoptosis induced by anti-DR5 mAb on synovial cells was measured by MTT analysis,DNA fragmentation and flow cytometry.Caspase-3 and Bcl-2 expression in synovial cells were detected by Western blot.The involvement of the apoptotic pathway was further proved by a caspase inhibition assay.Results:MTT result showed that Anti-DR5 mAb could inhibit synovial cells growth.And flow cytometry suggested that the cell death mode was apoptosis.The protein level of caspase-3 in synovial cells treated with anti-DR5 mAb was raised,while Bcl-2 level declined.When the caspase inhibitor was added to the synovial cells treated with anti-DR5 mAb,it was showed in a dose-dependence inhibition effect,indicating that anti-DR5 mAb inducing apoptosis might be through the caspase pathway.Conclusion:Anti-DR5 mAb could induce synovial cell apoptosis through caspase pathway.And this effect may be related to the protein level of Caspase-3 and Bcl-2.
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Objective: To construct the expressing vector of the extracellular domain of death receptor 5 (DR5), express it E.coli, identify the purified DR5 protein, and study its biological activity. Methods: The extracellular domain of DR5(eDR5) was assembled by overlapping PCR. The expression vector pET-22b(+)/ DR5 was constructed and transformed into E.coli BL21(DE3). The expression of eDR5 protein was induced by IPTG and purified by Ni 2+ -affinity chromatographic column. The purity and specificities were detected by SDS-PAGE and ELISA, respectively. The blocking effects of purified eDR5 on FMU1.5-induced apoptosis of U343, U373 cells were observed. Results: The extracellular domain of DR5 was obtained by overlapping PCR. The eDR5 protein was expressed in both supernatants and inclusion bodies with a yield more than 30% of total bacterial proteins. The purity of eDR5 was more than 95% and the yield reached 9 mg/ml. The result of ELISA showed the purified protein was eDR5. Purified eDR5 partially blocked the apoptosis of U343 cells induced by FMU1.5 and TRAIL. Conclusion: The successful construction, expression, and purification of the extracellular domain of DR5 protein lays a foundation for further study of DR5 function.
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Objective To investigate the in vitro anti-candidal activity of the essential oil of Illicium verum (EOIV) alone and in combination with fluconazole. Methods The minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of EOIV were determined in 130 clinically isolated Candida strains by NCCLS-M27-A microdilution method, and fluconazole was used as positive control. Meanwhile the checkerboard microdilution method was applied to assay the combined effect of EOIV and fluconazole in 18 candidal strains. Results For the 18 candidal strains the MICs and the MFCs of EOIV were 1 517.16 ?g/mL and 2 248.55 ?g/mL for C. albicans, 1 169.24 ?g/mL and 2 338.49 ?g/mL for C. glabrata, 1 320.03 ?g/mL and 1 741.79 ?g/mL for C. parapsilosis, 1 203.50 ?g/mL and 2 407.01 ?g/mL for C. tropicalis, 1 516.32 ?g/mL and 2 144.40 ?g/mL for C. krusei, and 1 072.64 ?g/mL and 2 144.40 ?g/mL for C. guilliermondii, respectively. Significant synergistic and additive effects were observed after the combination of EOIV with fluconazole, and no antagonism was found. There was no significant difference in the mean fractional inhibitory concentration index (FICI) between the fluconazole-susceptible and the fluconazole-resistant candidal strains (P = 0.671). Conclusion EOIV has antifungal effects on medically important Candida spp.. The combination of EOIV with fluconazole presents a synergistic and additive effects.
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Objective:To investigate possible correlation between nutritional status and forced expiratory volume in 1 second expressed as percent predicted (FEV1%) and serum levels of complement components C3 and C4 in patients with chronic obstructive pulmonary disease (COPD).Method:Body weight,FEV1% and serum levels of C3 and C4 in 27 healthy subjects and 40 stable COPD patients were determined and studied the correlation between them.Results:Body weight and C4 serum level in COPD group were much lower than that health group,while C3 serum level was no significant.A significant and positive correlation was present between FEV1% and decrease of body weight and C4 level(γ=0.517 9,0.527 3,respectively).Morever,there was significant correlation between body weight and C4 level (γ=0.487 3) in patients with COPD.Conclusion:In patents with stable COPD,pulmonary funciton was associated with the decrease of nutritional depletion (ND) and serum level of complement components C4,which was an indicator of showing the severe degree of COPD and prognosis.These results suggest that nutritional supplementation and immunological supporting treatment directly influence the curative effect of COPD.
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Objective:To express and purify a new fusion protein(RGD/tTF) for targeting therapy of cancer, and analyze its activities.Methods:The fused gene RGD/tTF was constructed by PCR, and then was inserted into vector pET22 b(+),expressed in E.coli BL21(DE3).The fusion protein was purified through Nickel-affinity chromatography column. The tTF activity of the fusion protein was analyzed by clotting assay and FⅩ activation assay. The specific binding of RGD/tTF to ?_v?_3 was analyzed by indirect ELISA.Results:The recombinant plasmid pET22 b(+)/RGD/tTF with correct sequence was obtained. The fusion protein was expressed with high yield in E.coli BL21(DE3). The purified fusion protein could activiate FⅩ and cause blood coagulation, and bind to ?_v?_3 specifically.Conclusion:The recombinant plasmid pET22 b(+)/RGD/tTF was constructed.The fusion protein retained TF activity and binding specificity to ?_v?_3, lays the foundation for studying the function of inducing thrombosis in tumor vasculature in vivo.
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Objective:To study the cytotoxic effects on tumor cell lines Hela,BGC823,MCF-7,L342,H9101,D6 induced by Fas ligand and anti-human DR5 monoclonal antibodies(Anti-DR5 mAb) and the underlying mechanism.Methods:Fas/DR5 mRNA were detected by RT-PCR. Cytotoxicity exerted by FasL/Anti-DR5 mAb on tumor cell lines was measured by MTT colorimetry and the induced apoptosis was determined by agarose gel electrophoresis.Results:The expression of DR5 on BGC823 and Hela cells were higher and DR5 didn’t express in D6. The expression of Fas on H9101 and L342 were higher and Fas didn’t express in D6. Cell line H9101 and L342 were sensitive to Anti-DR5 mAb and FasL in a dose dependent manner; Cell line MCF-7 and BGC823 were sensitive to FasL but were partially sensitive to Anti-DR5 mAb; Cell line Hela was partially sensitive to FasL but was sensitive to Anti-DR5 mAb; Cell line D6 was insensitive to two apoptosis inductions.Conclusion:Apoptosis induced by Fas ligand and Anti-DR5 mAb vary among tumor cell lines. The underlying mechanism may be relevant to Fas/DR5 mRNA expression,the release of Caspase-8 and Bcl-2.