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The aim of this study was to refold the OvisAries leukocyte antigen (OLA) class Ⅰ protein with peptides derived from sheeppox virus (SPPV) to identify SPPV T cell epitopes. Two pairs of primers were designed based on the published sequence of a sheep major histocompatibility complex Ⅰ to amplify the heavy chain gene of OLA Ⅰ α-BSP and the light chain gene of OLA Ⅰ-β2m. Both genes were cloned into a pET-28a(+) expression vector, respectively, and induced with ITPG for protein expression. After purification, the heavy chain and light chain proteins as well as peptides derived from SPPV were refolded at a ratio of 1:1:1 using a gradual dilution method. Molecular exclusion chromatography was used to test whether these peptides bind to the OLA Ⅰ complex. T-cell responses were assessed using freshly isolated PBMCs from immunized sheep through IFN-γ ELISPOT with peptides derived from SPPV protein. The results showed that the cloned heavy chain and light chain expressed sufficiently, with a molecular weight of 36.3 kDa and 16.7 kDa, respectively. The protein separated via a SuperdexTM 200 increase 10/300 GL column was collected and verified by SDS-PAGE after refolding. One SPPV CTL epitope was identified after combined refolding and functional studies based on T-cell epitopes derived from SPPV. An OLA Ⅰ/peptide complex was refolded correctly, which is necessary for the structural characterization. This study may contribute to the development of sheep vaccine based on peptides.
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Animals , Capripoxvirus , Epitopes, T-Lymphocyte/genetics , Peptides/genetics , Poxviridae Infections , Sheep , Sheep DiseasesABSTRACT
Objective To assess the effects of tacrolimus (FKS06) on podocyte in type 2 diabetic model rats and to explore the potential mechanism.Methods The model rats were fed with high fat and high sugar food and combining with a low-dose of streptozotocin (STZ).They were then randomly divided into a diabetic mellitus group (DM group) and a FK506 group.A normal control group (NC group) was also set.The rats in FK506 group were given with 0.5 mg· kg-1· d-1 FK506 for 8 weeks.The biochemical parameters were measured.The changes of renal pathology and ultrastructure of podocyte were observed by the light and electron microscopy.The expression of nephrin and LC3-Ⅱ was determined by immunohistochemistry and Western blotting.Results (1) Compared with those in NC group,KW/BW,systolic blood pressure (SBP),fasting blood glucose (FBG),triglyceride (TG),total cholesterol (TC),urinary albumin excretion rate (UAE) and creatinine clearance rate (Ccr) in DM group were significantly increased (all P < 0.05).And the KW/BW,UAE and Ccr were decreased in FK506 group compared to those in DM group (all P < 0.05),while other parameters had no significant difference (all P > 0.05).(2) Compared with those in NC group,the glomerular volume,mesangial cell proliferation and accumulation of mesangial matrix were increased,and the foot process became disorder and fusion in DM group,while these changes were significantly reduced in FK506 group.(3) Compared with that in NC group,the expression of nephrin and LC3-Ⅱ was decreased in DM group (all P < 0.05),and both of parameters were higher in FK506 group than those in DM group (all P < 0.05).Conclusion FK506 may enhance podocyte autophagy in type 2 diabetic model rats and attenuate podocyte injury.
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Objective To investigate the relationship between the infiltration of lymphocytes in renal tissues and podocytes injury in patients with lupus nephritis (LN),and provide the evidence of mechanism of podocytes injury in LN.Methods Thirty cases of biopsy proven LN patients were enrolled into the study,10 cases were selected as a the controls.The clinical and pathological data and renal tissues were collected.The infiltration of T lymphocytes (CD4+,CD8+ cells) and B lymphocytes (CD20+ cells) in renal tissues were detected by immunohistochemical method.The nephrin expression was detected by immunofluorescence.The relationship between CD4+,CD8+,CD20+ cells in renal tissues and 24 h UP,nephrin expression were analyzed by Pearson's correlation analysis,respectively.Results ① Compared with the control group,the 24 h UP was increased [2.86±1.37 vs 0.10±0.22 (g/24 h)];the infiltrations of inflammatory cells were increased significantly,the podocytes injury could be observed,combined with effacement of podocytes foot processes or disappeared in the LN group.Moreover,the increase of 24 h UP of active LN group was more evident than that in the inactive group [3.91 ±1.45 vs 1.77±0.69 (g/24 h),F=24.15,P<0.05],and the infiltration of inflammatory cellsand effacement of podocytes foot processes were more severe in the active LN group.②Compared with the control group,the nephrin expression decreased and the infiltration of CD4+,CD8+ and CD20+ cells in the renal tissues increased significantly,which was mainly aggregated in renal interstitial tissue.And In addition,CD4+ [98±13 vs 40±12 (cells/glomerulus),F=240.18,P<0.05],CD8+[109.0±16.4 vs 53.3±12.1(cells/glomerulus),F=210.40,P<0.05] and CD20+ [149.4±1.4 vs 82.6±13.3 (cells/glomerulus),F=544.30,P<0.05] cells of the active LN group were more remarkable than that in the inactive group.③ The Pearson correlation analysis showed that CD4+,CD8+ and CD20+ cells in renal tissues were positively correlated with 24 h UP (r=0.688,0.748,0.702;P<0.01),and were negatively correlated with nephrin expression (r=-0.623,-0.793,-0.693;P<0.01).Conclusion The infiltrations of CD4+,CD8+ and CD20+cells in renal tissues are very closely related with podocytes injury in patients with LN,and the infiltration of lymphocytes may be an important mechanism for podocytes injury in LN patients.
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Objective To examine the reliability and validity of the Connor-Davidson Resilience Scale (CD-RISC) in nurses in China.Methods Item analysis and confirmatory factor analysis of CD-RISC was conducted.A total of 1 084 nurses were randomly divided into 2 groups using random number table.One group was used to revise CD-RISC and the other to evaluate its reliability and validity.Results Confirmatory factor analysis of the original RSA showed thatx2/df,RMSEA,GFI,AGFI,CFI were 6.714,0.103,0.804,0.764,and 0.718,respectively.The number of item was reduced to 21 based on exploratory factor analysis results,including three factors which explained 54.034% of the total variance.Factor loading ranged from 0.490 to 0.755.Coefficient of internal consistency ranged from 0.688 to 0.930 and the 4 week test-retest reliabilities ranged from 0.825 to 0.917.The factor scores were closely correlated with the total scores with correlation coefficient ranging from 0.683 to 0.943.The factors were closely correlated with each other with correlation coefficient ranging from 0.564 to 0765.The confirmatory factor analysis of the revised CD-RISC showed that x2/df,RMSEA,GFI,AGF,CFI were 3.304,0.065,0.902,0.878,and 0.913 respectively.The total scores and all factors were negatively correlated with depression and anxiety and somatization.Conclusions The revised CD-RISC has good psychometric properties and can be used to measure the resilience of nurses.
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A new algorithm to generate the medical record report based on XML and linked list is proposed in this paper. It decomposes and abstracts the content of report. Reporter can search and reuse the resources quickly, when writing or modifying the report. The experiment results prove that the algorithm reduces the reporting time and standardizes the content of report.
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Humans , Algorithms , Medical Record Linkage , Medical Records Systems, Computerized , Programming LanguagesABSTRACT
To obtain the recombinant 18 kD protein with high purity and normal bioactivity of Cysticercus cellulosae (rCE18), E. coli cells with the rCE18 were disrupted ultra-sonically, and the inclusion bodies were washed with a solution containing 0.2% deoxycholic acid sodium (DOC)and 2% DOC, respectively. Then they were denatured with 0.9% sodium lauroyl sarcosine (SKL) followed by dialysis and gel filtration to refold and purify the target protein. At the same time, this method was compared with GST-FF affinity chromatography and recovering from SDS-PAGE gel. Biological activity of purified rCE18 was analyzed with indirect ELISA, and the purity of the products was identified using SDS-PAGE. The purity of refolded inclusion bodies exceeded 60% and the total recovery of activated protein rCE18 was about 41.3%. The specificity of rCE18 reached up to 97.2% using indirect ELISA. An effective way for purifying and refolding rCE18 expressed in E. coli as inclusion bodies was established, rCE18 with higher purity and activity was obtained, which has the potential for developing diagnosis methods of porcine cysticercosis.
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Animals , Antigens, Helminth , Genetics , Allergy and Immunology , Chromatography, Gel , Cysticercus , Genetics , Allergy and Immunology , Metabolism , Escherichia coli , Genetics , Metabolism , Inclusion Bodies , Metabolism , Protein Renaturation , Recombinant Proteins , Genetics , Allergy and ImmunologyABSTRACT
Objective To investigate the effect of positive ventilation pressure on the radiotherapy of primary cancer of stage Ⅲ.Methods 19 patients diagnosed as of primary,cancer of stage Ⅲ were randomly divided into two groups:the combining therapy group and the simple radiotherapy group.The patients of combining therapy group were treated with positive pressure ventilation using BIPAP respirator and radiotherapy.The recently therapy results and the radiotherapy associated side effects were observed between these two groups.Results(1)The combination of radiotherapy and BI- PAP provides significant superiority of local effects over radiotherapy.P
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Aim To study the pharmacokinetics of curcumin-pvp solid dispersion ig administration in mice in comparison with the free curcumin suspension.Methods Drugs were administered at a dose of 300 mg?kg-1 via ig.The plasma concentration of curcumin was determined by HPLC,the pharmacokinetics were calculated by DAS ver1.0 program.Results The curcumin pharmacokinetics conforms to a two-compartment open model after a single ig dose of curcumin solid dispersion in mice.The parameters were as follows:T12? and T12? were 16.4 and 266.1 min,respectively.AUC was 89.6 mg?L-1?min-1,Vd was 763.9 L,V1 was 51.0 L,and CL was 1.99 L?min-1.The absorption rate of curcumin solid dispersion was 6.75 times as much as curcumin suspension.Conclusion The curcumin solid dispersion improves the absorption of curcumin in vivo in mice evidently.
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AIM: To study the relation of acute radiation-induced oral mucositis to fungal infection and evaluate the effect of fluconazole in the treatment of patients with fungal infection. METHODS: Seventy-one patients (M 42, F 29; 52 a± s 41 a,11-76 a) with large area of oral mucosas irradiated were into this study. When most serious mucositis, the patients' oral mucosas were checked, 32 patients with fungal infection were treated with fluconazole (fluconazole 100 mg, po, qd×5 d or 150 mg,iv,gtt×3 d). RESULTS: The rate of fungal infection was 45%, the most of patients were infected by candida albicans. Degrees of mucositis between the patients with fungal infection to the patients without fungal infection were significantly different (P<0.05). The patients with fungal infection were treated with fluconazole. Degree of mucositis between the pretreated patients and post-treated patients were significantly different (P<0.05). CONCLUSION: The serious acute radiation-induced oral mucositis has significantly related with fungal infection. The patients with fungus infection treated with antifungal agents (fluconazole) can significantly relieve the reaction of acute radiation-induced oral mucositis.
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Objective To change the two-tiered C/S architecture of traditional PACS.Methods Based on the deeply research of PACS architecture,combination with DICOM and distributed object technology,propose distributed PACS system architecture,the three-tire client/server framework of distributed object technology introduced into the architecture of PACS to design the new architecture.Results In comparison with traditional PACS that were based on DICOM networks,the distributed PACS were improved themselves in system architecture and performance,and PACS were more open and interoperable.Conclusion The new architecture not only enhances the performance of PACS,but also meets the need of open and high interconnectivity and interoperability.
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53 cases of tumor of the head received radiotherapy during 1993 - 1995. Their tongue had received a rather high dosage of radial therapy. 44/53 cases had their tongue for thickened (P 0, 05). After radiotherapy, 36/53 cases had their tongue fur became scanty or even totally uncoated (P
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Objective:To evaluate the effect of different dietary pattern and soybean oligosac-charides supplementation on the amount and proportion of short-chain fatty acids (SCFA). Method: Twelve healthy students aged 20 to 25 years old were selected in the medical college. The study included 3 periods. In every period the students accepted different dietary patterns in 1st week [1. low animal food diet (LAFD),2. balanced food diet (BD), 3. high animal food diet (HAFD)]. Soybean oligosaccharides (5g/d) were added to different diets in 2nd week. The diet in 1st week was recovered in 3rd week. The study lasted for 9 w. Feces were collected once a week and SCFA was measured by capillary gas chromatography. Results: The total SCFA in feces were increased after taking LAFD, more prominent in acetic acid and butyric acid (P