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In this paper, two recombinant plasmids (pVIR-P12AIL18-3C and pVIR-P12A-3C) containing foot and mouth disease virus (FMDV) capsid polypeptide, 3C coding regions of O/NY00 and using/or not swine IL18 as a genetic adjuvant were constructed, and evaluated for their ability to induce humoral and cellular responses in mice and swine. In addition, the ability to protect swine against homologous virus challenge was examined. Mice and swine were given booster vaccination twice and once, respectively, and swine were challenged 10 days after the booster vaccination. Control groups were inoculated with pVAX1 or phosphate-buffered saline (PBS). All animals vaccinated with pVIR-P12AIL18-3C and pVIR-P12A-3C developed specific anti-FMDV ELISA antibody and neutralizing antibody and T lymphocyte proliferation and CTL cytotoxic activity was observed. In addition, we found that pVIR-P12AIL18-3C possessed stronger immunogenicity than pVIR-P12A-3C. The pVIR-P12AIL18-3C and pVIR-P12A-3C provided full protection in 3/4 and 2/4 swine from challenge with FMDV O/NY00, respectively. The results demonstrate the potential viability of a DNA vaccine in the control and prevention of FMDV infections.
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Adjuvantes Imunológicos , Proteínas do Capsídeo/imunologia , Cisteína Endopeptidases/imunologia , Vírus da Febre Aftosa/imunologia , Febre Aftosa/imunologia , Febre Aftosa/prevenção & controle , Interleucina-18/imunologia , Plasmídeos/imunologia , Vacinação , Proteínas Virais/imunologia , Vacinas Virais/imunologia , Proteases Virais 3C , Animais , Anticorpos Antivirais/sangue , Proteínas do Capsídeo/biossíntese , Proteínas do Capsídeo/genética , Divisão Celular , Cisteína Endopeptidases/biossíntese , Cisteína Endopeptidases/genética , Testes Imunológicos de Citotoxicidade , Ensaio de Imunoadsorção Enzimática , Feminino , Febre Aftosa/sangue , Imunização Secundária , Injeções Intramusculares , Interleucina-18/biossíntese , Interleucina-18/genética , Camundongos , Camundongos Endogâmicos BALB C , Testes de Neutralização , Plasmídeos/metabolismo , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/imunologia , Suínos , Linfócitos T/imunologia , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia , Proteínas Virais/biossíntese , Proteínas Virais/genética , Vacinas Virais/administração & dosagemRESUMO
Objective:To discuss the feasibility, safety and efficacy of kissing cover stents technique in the management of pseudoaneurysm near the bifurcation of innominate artery (the distance <1 cm).Methods:A retrospective study of 10 patients with pseudoaneurysm near the bifurcation of innominate artery were performed in the Affiliated Wuxi People’s Hospital of Nanjing Medical University from May 2013 to August 2018. There were 6 males and 4 females, aged from 25.0 to 77.0 years old, with a median age of 50.5 years old. All patients were treated with Viabahn stents combined with kissing stents. The adverse reactions were observed during the perioperative period, and the patients were followed up to evaluate the stents position and patency. Results:Among 10 patients, 2 were in the distal of innominate artery, 5 were in the bifurcation of innominate artery, 2 were in the root of right common carotid artery and 1 was in the origin of right subclavian artery. The distance from the injury site to the bifurcation of innominate artery was less than 5 mm in 4 cases and 8 mm in 1 case. All of them were successfully treated with Viabahn stents combined with kissing stents. Operation time ranged from 100.0 to 150.0 (122.0±14.9) min. The postoperative hospitalization time was (6.3±1.5) d, ranged from 4.0 to 9.0 d. During the operation, 8 mm×100 mm Viabahn stents were implanted in the right subclavian artery and 10 mm×100 mm Viabahn stents were implanted in the right common carotid artery. There were no severe adverse reactions during the perioperative period. Follow-up time ranged from 12 to 24 months. During the follow-up, color Doppler ultrasound and carotid CT angiography showed that the stents were in appropriate position, the blood flow was unobstructed, and there was no occlusion and displacement of stents. Viabahn stents remained at the level of proximal end of the innominate artery.Conclusion:Viabahn stents combined with kissing stents is a safe and effective treatment for pseudoaneurysm near the bifurcation of innominate artery.
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BACKGROUND:Previous studies have reported that rapamycin can affect the proliferation, migration and adhesion abilities of endothelial progenitor cels, but there is no report on the effect of autophagy, as wel as the interaction between autophagy and apoptosis. OBJECTIVE: To observe the effect of rapamycin activated autophagy activation on the proliferation, apoptosis, and cycle of endothelial progenitor cels. METHODS:Density gradient centrifugation was used to obtain mononuclear cels from bone marrow, and the mononuclear cels were inoculated on human fibronectin-coated culture plate.Then after cultured for 7 days the adherent cels colected were the endothelial progenitor cels. Different concentrations of rapamycin (0.01, 0.1, 1 and 10 μg/L) were added and cultured for 24 hours. Western blot was used to detect the LC3-II protein expression and monitor the induction of autophagy, flow cytometry was used to observe the cel cycle progression and apoptosis changes, and methylthiazolyldiphenyl-tetrazolium bromide colorimetric assay was used to observe the proliferation ability. Meanwhile, the ultrastructural changes were observed under transmission electron microscope. RESULTS AND CONCLUSION:Compared with the control group, there was no significant increasing of LC3-II protein expression of endothelial progenitor cels in 0.01 μg/L rapamycin group, and the LC3-II protein expression was in the high level. The LC3-IIprotein expression in the 1 μg/L and 10 μg/L rapamycin groups was higher than that in the control group, but lower than that in the 0.01 μg/L rapamycin group, which indicated that autophagywas particularly active when the concentration of rapamycin was 0.01 μg/L. The apoptosis of endothelial progenitor cels was increased with the increasing of concentration of rapamycin, and the proliferation rate was decreased with the increasing of concentration of rapamycin. The results indicate that activation of autophagy by bapamycin can promote the cel apoptosis, change the cel cycle significantly, and can inhibit the proliferation of endothelial progenitor cels.
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Objective To explore the expression of V-ATPase in colon cancer and its clinical significance. Methods Detecting the expression of V-ATPase mRNA in 20 paired of colon tumor tissues and normal tissues by using reverse transcription-polymerase chain reaction ( RT-PCR) and real-time quantitative polymerase chain reaction( Real-time PCR) , and testing the expression of V-ATPase protein by immu-nohistochemistry of EnVinsion. Results The expression of V-ATPase mRNA in tumor tissues and its paired normal tissues were (5. 37 ± 0. 44) and (2. 03 ± 0. 35)(P<0. 01). The positive immunohistochemistry of V-ATPase in tumor tissues and its paired normal tissues were 69. 1%(47/68) and 5. 8%(4/68) respectively, and the positive expression were primarily in cytoplasm and cytomembrane. Overexpression of V-ATPase was associated with tumor stage (P<0. 05), lymph node metastasis (P=0. 044), distant metastasis (P=0. 049), vessel in-vasion (P=0. 044) and differentiation (P<0. 001). Conclusion Overexpression of V-ATPase plays a significant role in the carcinogene-sis and the progression of colon cancer, which might be an important postoperative therapeutic target.
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Objective To evaluate the strategies and effect of surgical treatment for concomitant abdominal aortic aneurysm (AAA) and colorectal cancer (CRC). Methods Literatures on concomitant AAA and CRC published from January 1988 to December 2008 were retrieved from Pubmed, Sciencedirect, Ovid, CBMdisc, CNKI and et al, and correlated indexes were extracted for analysis. Differences among the groups were analyzed using the t test, chi-square test and fisher's exact test. Results A total of 367 cases of concomitant AAA and CRC treated by operation were retrieved. The length of operation delay of patients who received radical resection of CRC first was (115 ± 21 )days, which was significantly longer than (42 ± 8 )days of patients who received open abdominal aortic aneurysm repair (OAAR) first (t = 18. 9, P <0.05). The 30-day complication rate and accumulative length of hospital stay of patients who received one-stage radical resection of CRC + OAAR were 10.5% ( 12/114 )and (23 ±6) days, and 26.0% (47/181) and ( 16 ±4)days of patients who received two-stage radical resection of CRC + OAAR, with a significant difference ( χ2 = 10.42, t = 12. 01, P <0.05 ). The accumulative length of hospital stay of patients who received radical resection of CRC + endovascular aneurysm repair (EVAR) was (12 ±4) days, which was significantly shorter than that of patients who received radical resection of CRC + OAAR [ ( 19 ±5 ) days ] ( t = 9.48, P < 0. 05 ). The 4-year survival rate of patients who received two-stage radical resection of CRC + OAAR was 43.5% (27/62), which was significantly lower than that of patients who received two-stage radical resection of CRC + EVAR [69.2% (18/26) ] or one-stage radical resection of CRC + OAAR [73.7%(14/19) ] (χ2 =4.83, 5.28, P<0.05). Conclusions If the diameter of AAA is under 5 cm, radical resection of CRC should be firstly carried out; but if the diameter of AAA is above 5 cm, OAAR should be firstly carried out to prevent the rapture of tumors. One-stage surgery is better than two-stage surgery if patients could tolerate it.
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Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1)and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of infiuenza virus(H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus(H1N1)positiVe serum samples of mice.Results:Three Th/B cell epitopes containing HA_(73-87),HA_(125-139),HA_(188-205) were acquired Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus(H1N1)isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1)positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus(H1N1).Our study also establish the foundations for the further research of influenza virus infectlon and immunity mechanism,the recognition of influenza virus(H1N1)functional epitope and the development of epitope vaccines.
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We designed and constructed a fuse expression gene OAAT and staphylococcal enterotoxin A (SEA) on the basis of the OAAT designed and constructed which consists of the structural protein VP1 genes from serotypes A and O FMDV, 5 major VP1 immunodominant epitopes from two genotypes of Asia1 serotype, and 3 Th2 epitopes originating from the non-structural protein, 3ABC gene and structural protein VP4 gene. The recombinant plasmids pEA was constructed using SEA as a genetic adjuvant. Expressions of target gene from the pEA in Hela cell were verified by IFA and Western blotting. The experiment of BALB/c mice immunized with the DNA vaccines showed that pA and pEA could induce simultaneously specific antibodies against serotypes A, Asia1, and O FMDV, and the highest antibody titres were found in the pEA and inactivated vaccine groups compared to pA vaccinating mice. Compared with the control, the levels of IL-2, IFN-gamma, IL-4, and IL-10 expression by splenic lymphocytes from mice immunized with pA and pEA were significantly increased. In addition, we found that the levels of IL-2, IFN-gamma and IL-4 from the mice immunized with pEA was higher than mice immunized with pA did. The results of viral challenge in guinea pigs showed the pA, pEA and inactivated vaccine provided full protection in 2/4, 2/4, 3/4, 3/4 and 4/4, 4/4 guinea pigs from challenge with FMDV O/NY00 and Asial/YNBS/58, respectively. The results demonstrated fuse protein OAAT and SEA may be potential immunoge against FMDV, furthermore, SEA may be an effective genetic adjuvant for DNA vaccine.
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Animais , Humanos , Camundongos , Adjuvantes Imunológicos , Genética , Antígenos Virais , Alergia e Imunologia , Proteínas do Capsídeo , Genética , Alergia e Imunologia , Enterotoxinas , Genética , Alergia e Imunologia , Epitopos , Alergia e Imunologia , Febre Aftosa , Alergia e Imunologia , Vírus da Febre Aftosa , Alergia e Imunologia , Cobaias , Células HeLa , Camundongos Endogâmicos BALB C , Fragmentos de Peptídeos , Genética , Alergia e Imunologia , Vacinas de DNA , Alergia e Imunologia , Proteínas Estruturais Virais , Genética , Alergia e Imunologia , Vacinas Virais , Alergia e ImunologiaRESUMO
Objective To observe the effects of endovascular radiation (ER) on the proliferation and apoptosis of medial smooth muscle cells (SMC) and to discuss the possible mechanisms of radiation in the prevention of vascular restenosis (RS) in rabbits after carotid endarterectomy (CEA).Methods Forty rabbits undergoing CEA were randomly divided into four groups (each group=10) and given a radiation dose of 0, 10, 20 and 40 Gy 32 P respectively. Rabbits were killed on the 3rd, 7th, 14th, 28th and 56th day after operation. The specimens were collected and histopathologic examinations were done.Results Proliferation apparently occurred in the intima and media of carotid the lumen became narrow in the control group on the 14 th, 28 th and 56 th day after operation. While in the radiation groups, proliferation was apparently suppressed and the lumen was much less narrowed ( P
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Objective:To find the possible mechanisms of endovascular radiation in the prevention of restenosis in rabbits following carotid endarterectomy and to observe the dose effect relationship.Methods:Twenty four rabbits were randomly divided into three groups following carotid endarterectomy (each n=8) and were allocated to receive a radiation dose of 0,10,20Gy( 32 P)respectively.Changes in the levels of plasma nitric oxide(NO) and endothelin 1(ET 1)were measured 3 day before operation and 3,7,14,28 day after operation.Results:The plasma NO was markedly increased and ET 1 was markedly decreased in endovascular radiation groups compared to control groups.The results were similar in the two groups(10Gy versus 20Gy).Conclusion:The endovascular radiation may inhibit smooth muscle cell proliferation and migration,and thus prevent restenosis by increasing NO and decreasing the level of ET 1.Changes of the levels of plasma NO and ET 1 is a practicable methods to detect the effect of therapy.
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Objective:To predict Th/B cell epitopes in HA of influenza virus(H1N1) and analyze antigenicity of the candidate epitopes in order to develop epitope-bacterin by the way of bioinformatics.Methods:The HA amino acid sequences of influenza virus (H1N1),which the viral infection was prevalent recently,were downloaded from Genbank.The Th/B cell epitopes were predicted and analyzed by bioinformatics methods.Then,specificity and conservation of the candidate epitopes were estimated.Finally,antigenicity of the candidate epitopes was identified by influenza virus (H1N1) positive serum samples of mice.Results:Three Th/B cell epitopes containing HA73-87,HA125-139,HA188-205 were acquired.Two of the candidate epitopes were in a relatively conserved domain of HA1,and a deal of 2006-2009 influenza virus (H1N1) isolates contained the sequences.Moreover,the candidate epitopes were showedin a distinct antibody combining reactivity with the influenza virus (H1N1) positive serum of mice,which inferred the predicted epitopes to be functional ones.Conclusion:The selected epitopes are able to be functional HA Th/B cell epitopes of influenza virus (H1N1).Our study also establish the foundations for the further research of influenza virus infection and immunity mechanism,the recognition of influenza virus (H1N1) functional epitope and the development of epitope vaccines.