Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 65
Filtrar
1.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-883092

RESUMO

Objective:To investigate the influence of peer education based on WeChat support on the social support, mental flexibility and rehabilitation self-efficacy of patients after laparoscopic radical prostatectomy.Methods:A total of 82 patients after laparoscopic radical prostatectomy who were admitted from January 2018 to December 2019 were selected as the research objects. According to the operation time, they were divided into the treatment group (January 2019-December 2019) with 43 cases and the control group (January 2018-December 2018) with 39 cases. The control group was given regular health education, and the treatment group jointly applied peer education based on WeChat support. Followed up for 2 months, the two groups of patients were evaluated the degree of social support, psychological flexibility, and rehabilitation self-efficacy by Social Support Rating Scale, Connor-Davidson Resilience Scale, General Self-Efficacy Scale and compared.Results:There was no significant difference in the degree of social support, psychological flexibility, and rehabilitation self-efficacy before intervention between the two groups( P>0.05). The objective support, support utilization, and social support scores in the treatment group after intervention were (9.12±1.12), (10.45 ± 0.75), (32.49 ± 4.56) points, and the control group were (7.45 ± 1.36), (8.74 ± 1.43), (29.84 ± 4.45) points, and the differences were statistically significant ( t values were 6.091, 6.681, 2.658, P<0.01). The scores of toughness, self-improvement, optimism, and mental resilience in the treatment group after intervention were (28.21 ± 4.25), (20.32 ± 3.54), (9.36 ± 1.12), and (57.89 ± 7.21) points, and the control group were (24.36 ± 4.34), (17.14 ± 3.21), (7.84 ± 1.23), (49.34 ± 6.55) points, and the differences were statistically significant( t values were 4.056-5.857, P<0.01). The scores of physical exercise self-efficacy, coping self-efficacy, and rehabilitation self-efficacy in the treatment group after intervention were (43.43 ± 5.38), (54.45 ± 6.32), (97.88 ± 7.45) points, and the control group were (37.45 ± 5.42), (48.65 ± 6.45), (86.10 ± 9.12) points, and the differences were statistically significant ( t values were 5.009, 4.110, 6.430, P<0.01). Conclusions:Peer education based on WeChat support helps to enhance the degree of social support for patients after laparoscopic prostate cancer surgery, improve the level of mental flexibility, and promote the development of rehabilitation self-efficacy.

2.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-806318

RESUMO

Objective@#To investigate the cellular and humoral immune responses induced by combined immunization with the fusion protein of human papillomavirus type 18 (HPV18) and the recombinant vaccinia virus.@*Methods@#Purified HPV18L231-600E7E6 fusion protein, expressed by prokaryotic expression system, were immunized in combination with the recombinant vaccinia virus vaccine expressing HPV18E7E6 fusion protein (rVV18E7E6) by using various prime-boost regiments in C57BL/6 mice. Cellular and humoral immune responses were analyzed by enzyme-linked immunospot assay (ELISPOT), enzyme-linked immunosorbent assay (ELISA), and pseudovirus neutralization assay.@*Results@#Higher levels of cellular immune responses were induced in mice primed with the HPV18L231-600E7E6 fusion protein/adjuvant CpG and boosted with the recombinant vaccinia virus rVV18E7E6, than in other immunized mice. Higher binding antibody level was induced, and low level neutralizing antibody against pseudovirus was detected simultaneously.@*Conclusions@#Priming with HPV18L231-600E7E6 fusion protein/CpG and boosting with the recombinant vaccinia virus rVV18E7E6 could induce higher cellular and humoral immune response in immunized mice, which might be taken as vaccine candidate for treatment of HPV18 chronic infection and postoperative adjuvant treatment for cervical cancer.

3.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-806032

RESUMO

Objective@#To detect the expression level of early and late protein of vaccinia virus and to preliminarily explore replication-defective mechanism of highly attenuated NTV strain of vaccinia virus Tiantan.@*Methods@#We constructed prokaryotic expression vector, expressed and purified homologous early protein E3 and late protein A27 closely related to replication and prepared mouse polyclonal antiserum by immunizing mice with homologous proteins. Early and late protein expression levels of NTV were detected.@*Results@#We have expressed and purified vaccinia virus proteins respectively in E. coli expression system and prepared homologous mouse polyclonal antiserum. Early protein E3 and late protein A27 could be highly efficient expression in NTV infected non-permissive Hela cells, while expression of late protein F17 was blocked detected by Western blot.@*Conclusions@#The expression limitation of late protein F17 may be an explanation for the replication-defective mechanism of NTV.

4.
Chinese Traditional Patent Medicine ; (12): 1009-1014, 2018.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-710259

RESUMO

AIM To study the reproductive toxicity of Triptolide,one of the main components of Tripterygium wilfordii Hook.f,on male Caenorhabditis elegans,and its potential mechanism.METHODS Expose L4 larva of him-5 nematodes to 48 h 0.02,0.2,2.0 mg/L of Triptolide.The effect of Triptolide on the fertility of male C.elegans was assessed by crossing test,and the relevant mechanism was explored through measuring spermatids morphology development,spermatids activation,sperm motility and the relative mRNA expression levels of the related spermatogenesis genes.RESULTS Compared with the solvent control group,the 48 h Triptolide treatment induced significantly decreased brood size in 2.0 mg/L Triptolide group,significant reduction in spermatid diameter,spermatid cross-sectional area,rates of spermatid activation,rates of motile sperm in 0.2 and 2.0 mg/L Triptolide groups,and decreased expression levels of spe-10 in 0.02-2.0 mg/L Triptolide groups,decreased spe-15 in 0.2-2.0 mg/LTriptolide groups,and decreased fer-1 and folt-1 in 2.0 mg/L Triptolide groups as well.CONCLUSION Triptolide decreases expression levels of the related genes in spermatogenesis,contributes to an array of indicators inhibition,the spermatids morphology development,the sperm morphology and function development,which leads to decreased fertility of male C.elegans.Therefore,spermatogenesis damage is one of the main pathway that Triptolide adversely affects the fertility.

5.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-808469

RESUMO

Objective@#To prepare strains of influenza A (H7N9) pseudovirus derived from different districts of China for vaccine efficacy evaluation.@*Methods@#Phylogenetic tree was built based on hemagglutinin (HA) amino acid sequence analyses from 29 influenza A (H7N9) virus strains and 6 influenza A (H7N9) virus strains with HA determinants variation were selected. 293FT cells were co-transfected with plasmid pNL4-3-Luc.R-E-, pVRC-HA and pVRC-NA with codon-optimized hemagglutinin (HA) and neuraminidase (NA) derived from the six influenza A (H7N9) virus strains, respectively. Transmission electron microscopy assay and Western blot analysis were performed to demonstrate morphology and specificity of these particles, luciferase activity assay and hemagglutinin titers detection were used to determine their infectivity and hemagglutinin activity. And finally, pseudovirus-based neutralization assays were evaluated with HA immunized mice serum.@*Results@#Six influenza A (H7N9) peseudovirus particles derived from different districts of China were selected and prepared. All of the particles bearing HA and NA were characterized with classic influenza virus morphology, with TCID50 titer ranged from 104TCID50/50 μl to 105TCID50/50 μl and with hemagglutinin activity ranged from 64 to 512. Neutralization efficacies on influenza A/Shanghai/1/2013(H7N9) HA vaccine serum against 100TCID50 dose of these pseudovirus particles indicated their potential application in the vaccine cross-protective evaluation in future.@*Conclusions@#Six influenza A (H7N9) pseudovirus derived from different districts of China with potential antigenic variation on HA were constructed successfully, established foundation for their further application in vaccine cross-reactive efficacy evaluation.

6.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-808156

RESUMO

Objective@#To generate monkeypox virus specific monoclonal antibodies for further establishing monkeypox virus immunofluorescence assay.@*Methods@#Monkeypox virus A29 protein, vaccinia ortholog A27 protein and cowpox ortholog 162 protein were expressed in E. coli BL21 to screen antibodies. Synthetic monkeypox virus A2917 ~ 49 polypeptide was used to immune BALB/c mice. Monkeypox virus monoclonal antibodies were generated through fusion, cloning and screening techniques. Indirect ELISA was performed to test antibodies specificity and subtype.@*Results@#A29, A27 and 162 proteins were highly expressed in E. coli and detected by Western blot. The three his-tagged proteins were purified using His-Bind affinity chromatography column. The purity of the proteins was all more than 90%. And 8 strains monkeypox virus specific monoclonal antibodies were screened by the three purified proteins. Two mAbs of 8 were IgG3 subtype and the rest were IgG1 subtype.@*Conclusions@#Eight strains of monkeypox virus specific monoclonal antibodies were generated, they can be used to further establish monkeypox virus immune immunofluorescence assay.

7.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-660943

RESUMO

Objective To compare the value of shear wave elastography (SWE)and mammography in differential diagnosis of benign and malignant breast neoplasms.Methods Totally 96 patients with breast tumor were randomly chosen and underwent SWE andmammography.The elastic maximum value (Emax)was obtained. Taking histological diagnosis as the golden standards,we compared the two techniques' sensitivity,specificity, accuracy,positive and negative predictive value in diagnosis of breast tumors.Results The sensitivity,specificity, accuracy,positive and negative predictive value of Emax were 91.3%,94.0%,92.7%,93.3% and 92.1%, respectively.The sensitivity and accuracy of E-max were significantly better than those of x-ray (both P <0.05 ). The area under the ROC curve of Emax was 0.983 (95% CI,0.963 to 0.998).Conclusion SWE outperforms mammography in differential diagnosis of benign and malignant breast tumors.

8.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-658155

RESUMO

Objective To compare the value of shear wave elastography (SWE)and mammography in differential diagnosis of benign and malignant breast neoplasms.Methods Totally 96 patients with breast tumor were randomly chosen and underwent SWE andmammography.The elastic maximum value (Emax)was obtained. Taking histological diagnosis as the golden standards,we compared the two techniques' sensitivity,specificity, accuracy,positive and negative predictive value in diagnosis of breast tumors.Results The sensitivity,specificity, accuracy,positive and negative predictive value of Emax were 91.3%,94.0%,92.7%,93.3% and 92.1%, respectively.The sensitivity and accuracy of E-max were significantly better than those of x-ray (both P <0.05 ). The area under the ROC curve of Emax was 0.983 (95% CI,0.963 to 0.998).Conclusion SWE outperforms mammography in differential diagnosis of benign and malignant breast tumors.

9.
J Cancer Res Ther ; 11(2): 485-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26148625

RESUMO

To analyze the therapeutic effect of bevacizumab on radiation-induced brain necrosis. Four radiation-induced brain necrosis patients, administered with bevacizumab at a dose of 7.5 mg/kg every 3 weeks, 2 times. One case of brain metastasis of lung cancer and one case of nasopharyngeal carcinoma with brain necrosis after radiotherapy. However, their physical signs disappeared after the treatment with bevacizumab. One case of brainstem lesion and one case of brain glioma patient showed a transient improvement in signs and symptoms after treatment with bevacizumab. Bevacizumab can significantly alleviate the radiation-induced brain edema, and can improve the symptoms successively.


Assuntos
Inibidores da Angiogênese/uso terapêutico , Bevacizumab/uso terapêutico , Neoplasias Encefálicas/tratamento farmacológico , Encéfalo/efeitos dos fármacos , Necrose/tratamento farmacológico , Lesões por Radiação/tratamento farmacológico , Adulto , Encéfalo/patologia , Neoplasias Encefálicas/patologia , Carcinoma , Criança , Feminino , Glioma/patologia , Humanos , Neoplasias Pulmonares/patologia , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/patologia , Necrose/patologia , Lesões por Radiação/patologia
10.
Chinese Journal of Virology ; (6): 554-560, 2014.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-280328

RESUMO

To compare different adjuvant formulation and explore the impact of Calcineurin B subunit(CnB) as adjuvant with a novel HBV protein particle (HBSS1) vaccine in mice, female C57BL/6 mice were immunized HBSS1 with Al(OH)3 only, or a normal dose (5 μg) CnB only, or (CnB+ Al(OH)3) mixture as the adjuvant. All immunized groups were primed twice at 4-week intervals; followed by boosting with recombinant adenoviral based HBV vaccine(rAdSS1) at 10-week intervals. We detected the antigen specific humoral response in mice, including total IgG antibody and IgG subtyping. Then, we characterized the specific cell-mediated immune (CMI) response by detection of γ-interferon secreting splenocytes after stimulaton with S or PreS1 peptide pools. No enhancement of immunity was found among the mice with 5 μg of CnB alone or combined with Al(OH), adjuvanted vaccine,which could not induce higher level of anti-PreS1 and anti-S antibodies and CMI than that of HBSS1 alone or Al(OH)3 adjuvanted vaccines. We concluded that CnB is not an effective adjuvant for a novel HBV subunit vaccine.


Assuntos
Animais , Feminino , Camundongos , Adjuvantes Imunológicos , Farmacologia , Calcineurina , Farmacologia , Vacinas contra Hepatite B , Alergia e Imunologia , Camundongos Endogâmicos C57BL , Subunidades Proteicas
11.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-318091

RESUMO

<p><b>OBJECTIVE</b>To express HPV31 and 52 L2 fusion protein and detect its immunogenicity.</p><p><b>METHODS</b>According to the amino acid sequences of HPV31 and 52 L2 11-200AA published in the GenBank database, weartificially synthesized the HPV31 and 52 L2 fusion gene which was optimized according to Escherichia coli codon usage and encodes 11-200 amino acid of HPV31 and HPV52 L2, then cloned it into pET-9a vector. The HPV31 and 52 L2 fusion protein was expressed in Prokaryotic expression system and the mice were immunized with the fusion protein after purification. The immunogenicity was characterized in vaccinated mice.</p><p><b>RESULTS</b>HPV31 and 52 L2 fusion protein was highly expressed in E. coli, the amount of fusion protein is nearly 20% of the total bacterial protein. The purified fusion protein with aluminum adjuvant could induce specific high titer of IgG antibodies detected by ELISA, and also induce the neutralizing antibodies against pseudovirus of HPV31 and HPV52 and cross-neutralizing antibodies against pseudovirus of HPV45, 58, 16, 18.</p><p><b>CONCLUSION</b>HPV31 and 52 L2 fusion protein could induce neutralizing and cross-neutralizing antibodies against HPV pseudovirus. It provides laboratory basis for development of HPV L2 protein vaccine.</p>


Assuntos
Animais , Feminino , Camundongos , Anticorpos Antivirais , Sangue , Escherichia coli , Genética , Camundongos Endogâmicos BALB C , Proteínas Oncogênicas Virais , Genética , Alergia e Imunologia , Papillomaviridae , Alergia e Imunologia , Vacinas contra Papillomavirus , Alergia e Imunologia , Proteínas Recombinantes de Fusão , Alergia e Imunologia
12.
Chinese Journal of Oncology ; (12): 28-32, 2013.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-284245

RESUMO

<p><b>OBJECTIVE</b>To map the frequency and types of EGFR gene mutations present in lung cancer tissues. To evaluate the clinical applicability of a novel real-time double-loop probe PCR of which the ADx-EGFR kit is based, and to compare its performance with traditional Sanger DNA sequencing in the detection of somatic mutations of tumor genes.</p><p><b>METHODS</b>A total of 208 formalin-fixed paraffin-embedded (FFPE) tumor samples were tested. Genomic DNA of the tissue samples was extracted and purified, and subjected to both traditional PCR amplification, Sanger sequencing of EGFR gene in exon 18, 19, 20, 21, and ADx's EGFR mutation detection kit. The mutation rates for EGFR gene in exon 18, 19, 20, 21, as well as the frequency of each mutation detected by the two methods, were analyzed.</p><p><b>RESULTS</b>The traditional Sanger DNA sequencing technique was successfully performed in 196 out of 208 (94.2%) lung cancer samples, and 22 samples (11.2%) showed EGFR gene mutations. ADx-EGFR kit was successfully used in the lung cancers of all of the 208 cases (100.0%), and 40 samples (19.2%) showed mutations. In the lung cancer samples analyzed, mutations were mainly detected in the exon 19 and exon 21 L858R point mutation, i.e. 4.8% (10/208) and 11.6% (23/208) of total mutations, respectively, and the remaining mutations were rare.</p><p><b>CONCLUSIONS</b>The success rate of ADx-EGFR real-time PCR for formalin-fixed and paraffin-embedded tissues samples is significantly higher than that of Sanger sequencing (P < 0.01). There are significant differences between the two methods. ADx-EGFR real-time PCR shows a much higher successful detection rate and mutation rate of lung cancer tissues compared with that of Sanger sequencing. As a result, the real-time PCR with ADx-EGFR kit is proved to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing. It is an effective and reliable tool for clinical screening of somatic gene mutations in tumors.</p>


Assuntos
Humanos , Análise Mutacional de DNA , Métodos , Éxons , Genes erbB-1 , Neoplasias Pulmonares , Genética , Inclusão em Parafina , Mutação Puntual , Reação em Cadeia da Polimerase em Tempo Real , Métodos
13.
Journal of Leukemia & Lymphoma ; (12): 484-486, 2012.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-472326

RESUMO

Objective To investigate the influence of indomethacin to the unite treatment effect on chronic phase myeloid leukemia (CML-CP) with interferon alpha-2b (IFNα-2b) and low dose cytarabine (LD-Ara-C).Methods 22 CML-CP patients were randomly divided into two groups.Control groups (10 cases) injected with IFNα-2b (3 million units),injection frequency was q.o.d,the duration of treatment was about 3-18 months,cytarabine (Ara-C) by slowly intravenous driped (30 mg/d).In this treatment schedule,every course of treatment sustained 10 days,and with a 2 weeks interval.During this process,patients in treatment group were treated with hydroxyl urea only when their WBC in peripheral blood exceed 20×109/L,otherwise,discontinue it.Treatment group (12 cases),on the first day of treatment with IFNα-2b and Ara-C,added indomethacin (25 mg) through oral administration,the frequency was t.i.d.During treatment in the two group,the end point of observation was completely hematology ease,at the same time,these indicators in the two groups needed to be compared,the time when WBC begin to fall,the time when WBC fall to normal range,the time when immature cells returned to normal,the time which complete hematological remission and the highest temperature of patients after IFNα-2b was subcutaneous injected.Results The time when WBC begin to fall in treatment group was (4.2±2.7) d,and the time was (5.0±2.5) d in control group (t =0.714,P > 0.05).In treatment group,the time when WBC fall to normal range was(10.0±4.5) d,and the time was (12.0±4.5) d in control group (t =1.036,P > 0.05).The time when immature cells returned to normal in treatment group was (14.2±4.8) d,and the time was (19.0±3.6) d in control group (t =2.609,P < 0.02).The time which complete hematological remission was achieved in control group was (45.8±5.6) d,but it was (53.9±10.5) d in control group (t =2.314,· P < 0.05).Meanwhile,the fever degree after IFNα-2b was subcutaneous injected obviously achieved improvement in treatment group (x2 =12.041,P < 0.005).Conclusion The advantage of indomethacin to the unite treatment with IFNα-2b and LD-Ara-C on CML mainly lays in which cound alleviated the adverse reaction such as flu-like of IFN,and more,there are synergy effect in antagonist CML.

14.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-428757

RESUMO

ObjectiveTo construct one recombinant adenovirus AdE7E6 expressing HPV16 E6 and E7 fusion protein as candidate for HPV16 therapeutic vaccine.MethodsThe codon-optimized E6 and E7 gene,were fused to create one open reading frame,then inserted into adenovirus vector pCD316.A strain of recombinant adenovirus was constructed through homologous recombinant in 293 cells,and identified by PCR and Western blot.Finally,it was employed to study it's immunogenicity and the activity of the tumor growth regression.ResultsThe PCR result showed that E6E7 fusion gene had been integrated in recombinant Ad5 DNA.Western blot test confirmed that the E6E7 fusion protein was highly expressed in 293 cells infected with Ad5E7E6 recombinant adenovirus.The recombinant adenovirus elicited significant E7 specific CD8+ T lymphocyte response in vaccinated mice.These responses could completely prevent the TG-1 tumor cell bearing mice treated with AdE7E6 from developing into tumor.ConclusionThese results suggested that rAd5E7E6 could be a potent vaccine candidate for the treatment of HPV16-associated tumors and their precancerous transformations.

15.
Chinese Journal of Oncology ; (12): 810-815, 2012.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-307288

RESUMO

<p><b>OBJECTIVE</b>To investigate the high expression of HPV16L2N120E7E6 fusion protein by prokaryotic expression system, and evaluate its immunogenicity and antitumor efficacy in vaccinated mice.</p><p><b>METHODS</b>The HPV16L2N120E7E6 fusion gene, its codons were optimized to increase the expression of the protein, was constructed by overlap extension PCR and inserted into prokaryotic expression vector pET9a. Then the fusion protein was expressed by inducing with IPTG in E. coli strain BL21 (DE3) harboring with plasmid pETL2N120E7E6, and further detected by SDS-PAGE and Western-blot. Finally, the humoral and cellular immune responses were measured by ELISA and ELISPOT, respectively, in vaccinated mice with the purified HPV16L2N120E7E6 fusion protein, and the antitumor efficacy was assessed in mice using the TC-1 tumor challenge model.</p><p><b>RESULTS</b>The codon-optimized HPV16L2N120E7E6 fusion gene was highly expressed in E. coli strain BL21 (DE3) harboring with plasmid pETL2N120E7E6, and the amount of fusion protein was nearly 48.6% of the total bacterial protein. The purified fusion protein could induce high titer of specific antibody against L2, E7 and E6 in vaccinated mice. When accompanied with the adjuvant CpG, the fusion protein was able to elicit strong and moderate cellular immune responses in vaccinated mice against peptide HPV16E7(49-57) and peptide pools of HPV16E6, respectively. Furthermore, the tumor therapeutic experiment showed that HPV16L2N120E7E6 + CpG could prevent the tumor formation in 80.0% (8/10) vaccinated mice.</p><p><b>CONCLUSIONS</b>The data of this study suggest that HPV16L2N120E7E6 fusion protein could be a promising candidate vaccine for treatment of chronic HPV16 infection and post-operative adjuvant therapy for cervical cancer.</p>


Assuntos
Animais , Feminino , Humanos , Camundongos , Adjuvantes Imunológicos , Farmacologia , Vacinas Anticâncer , Alergia e Imunologia , Usos Terapêuticos , Proteínas do Capsídeo , Genética , Alergia e Imunologia , Metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Códon , Escherichia coli , Alergia e Imunologia , Metabolismo , Imunização , Métodos , Imunoterapia , Métodos , Camundongos Endogâmicos C57BL , Transplante de Neoplasias , Oligodesoxirribonucleotídeos , Alergia e Imunologia , Proteínas Oncogênicas Virais , Genética , Alergia e Imunologia , Metabolismo , Proteínas E7 de Papillomavirus , Genética , Alergia e Imunologia , Metabolismo , Vacinas contra Papillomavirus , Alergia e Imunologia , Usos Terapêuticos , Plasmídeos , Proteínas Recombinantes de Fusão , Genética , Alergia e Imunologia , Metabolismo , Proteínas Repressoras , Genética , Alergia e Imunologia , Metabolismo
16.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-246187

RESUMO

<p><b>OBJECTIVE</b>To determine the antigen characteristics of different fragments of SARS-CoV N protein expressed in E. Coli and their application in the serological diagnosis.</p><p><b>METHODS</b>Based on preliminary analysis of 39 different segments of the N protein, We choosed six purified N protein for further antigenicity characterization in this study, including that PN360 (1 -360aa), PN301 (1-301aa), PN199 (30-228aa), PN185 (30-214aa), PN155b (60-214aa), and PN125 (90-214aa). We developed Western-Bolt and ELISA to detect antibody reactivity between truncated N fragments with sera from SARS-CoV-negative normal adults or SARS-CoV patient convalescent sera.</p><p><b>RESULTS</b>Western-Bolt results show that all the six fragments have reacted with the SARS patient convalescent sera, but the PN360 and PN301 showed obvious cross-reaction with sera from SARS-CoV-negative normal adults; sensitivity analysis using an ELISA coating with PN199, PN185, PN155b, PN125 as antigen showed that the PN185 and PN155b are better than PN125.</p><p><b>CONCLUSION</b>Truncated N protein PN185 and PN155b expressed in E. Coli are better antigen candidates used for detection of SARS-CoV specific antibody.</p>


Assuntos
Humanos , Anticorpos Antivirais , Sangue , Western Blotting , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Genética , Proteínas do Nucleocapsídeo , Alergia e Imunologia , Fragmentos de Peptídeos , Alergia e Imunologia , Proteínas Recombinantes , Alergia e Imunologia , Testes Sorológicos , Síndrome Respiratória Aguda Grave , Diagnóstico
17.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-351529

RESUMO

To develop novel and effective HBV therapeutic vaccine, we constructed an expression vector, pVRC-HBSS1, in which PreS1 (21-47aa) coding gene fused to the C-terminal of the S (1-223 aa) coding gene of HBV, and prepared the protein particle vaccine HBSS1 that consist of S and PreS1 fusion antigen derived from CHO system. We immunized mice by priming three times with DNA vaccine via different methods (i.e., intramuscular injection, intradermal injection with electroporation), then boosting once with protein particle vaccine. We analyzed the immune response among various vaccination groups. The higher level of S or PreS1 specific antibodies was detected in the group via intradermal injection with electroporation, compared with that of direct intramuscular injection. We further found that the specific cellular immune responses (IFN-gamma ELISpot analysis) in the group priming with DNA vaccines and boosting with protein subunit vaccine particles, was significantly higher than that of the DNA or protein particle subunit alone. Moreover, combination vaccination priming with intradermal injection DNA via electroporation and boosting with protein particle induced the strongest cellular immune response. These results provide a basis for rational design and application of the novel HBV therapeutic vaccine.


Assuntos
Animais , Feminino , Humanos , Camundongos , Eletroporação , Fusão Gênica , Antígenos de Superfície da Hepatite B , Genética , Alergia e Imunologia , Vacinas contra Hepatite B , Alergia e Imunologia , Imunidade Celular , Alergia e Imunologia , Imunização Secundária , Camundongos Endogâmicos BALB C , Precursores de Proteínas , Genética , Alergia e Imunologia , Vacinas de DNA , Alergia e Imunologia , Vacinas de Subunidades , Alergia e Imunologia , Proteínas do Envelope Viral , Genética , Alergia e Imunologia
18.
Chinese Journal of Virology ; (6): 594-598, 2011.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-354786

RESUMO

This study aimed to develop an effective experimental vaccine against highly pathogenic H5N1 Avian Influenza (HPAI) virus and to optimize their immunization programs. As reported previously, various DNA-based or recombinant vaccinia viral(Tiantan)-based H5N1 vaccine candidates, which containing a single cistronic construct (HAop, or NAop) or a bicistronic construct (HAop/M2 or NAop/M1) of H5N1 influenza virus (Anhui strain) were constructed and characterized in our lab. In this study, we further analysed the immunogenicity in mice of these vaccine candidates by various protocols (single or combined immunization). Our results showed that: comparing with immunization with DNA-based or rTTV-based H5N1 vaccine only, combined DNA-based with rTTV-based H5N1 vaccine immunization via prime-boost strategy enhanced immune response significantly against multi-H5N1 antigens detected by hemagglutination inhibition (HI) assay, NA- or M1- or M2-specific antibody detection, and micro-neutralizing antibody test and IFN-gamma ELISpot assay. Priming with DNA-based vaccine induced higher level of humoral response against HA or NA antigen than priming with rTTV-based vaccine; In contract, M1 and M2-specific antibody levels were higher among that of priming with rTTV -based vaccine. These findings provide a basis for further development of novel H5N1 vaccines and for the optimization of the immunization programs of combined multi-antigens vaccine candidates.


Assuntos
Animais , Feminino , Camundongos , Antígenos Virais , Genética , Alergia e Imunologia , Imunização , Métodos , Virus da Influenza A Subtipo H5N1 , Genética , Alergia e Imunologia , Vacinas contra Influenza , Genética , Alergia e Imunologia , Camundongos Endogâmicos BALB C , Vacinação , Métodos , Vacinas de DNA , Genética , Alergia e Imunologia , Vacinas Sintéticas , Genética , Alergia e Imunologia , Vaccinia , Genética , Alergia e Imunologia
19.
National Journal of Andrology ; (12): 880-883, 2011.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-305772

RESUMO

<p><b>OBJECTIVE</b>To investigate the relationship between vitamin D receptor (VDR) gene Fok I polymorphisms and BPH complicated by histological prostatitis (BPH + HP).</p><p><b>METHODS</b>We detected Fok I polymorphisms in the peripheral blood of 79 patients with BPH + HP and 81 controls with BPH only using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), and analyzed the frequency distribution of the genotypes and alleles of the two groups of patients.</p><p><b>RESULTS</b>Obvious differences were found in the distribution of genotypes and alleles between the BPH + HP patients (FF: 27% [21/79], Ff: 30% [24/79], ff: 43% [34/79]) and the controls (FF: 33% [27/81], Ff: 36% [29/81], ff: 31% [25/81]), with statistical significance in the distribution of the genotype ff (P < 0.05). The histological prostatitis group showed a significant difference from the controls in the frequency of the f allele (58% [196/338] vs 49% [153/312], P < 0.05), but not in that of the F allele (42% [142/338] vs 51% [159/312] , P > 0.05).</p><p><b>CONCLUSION</b>VDR Fok I polymorphisms may be correlated with BPH complicated by histological prostatitis.</p>


Assuntos
Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Alelos , Estudos de Casos e Controles , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único , Hiperplasia Prostática , Genética , Prostatite , Genética , Receptores de Calcitriol , Genética
20.
Chinese Journal of Virology ; (6): 34-43, 2011.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-286081

RESUMO

The purpose is to screen and identify the specific H-2d restricted T-cell epitopes. These epitopes are used to investigate the cellular immune response of BALB/c (H-2d) mice immunized with a HIV-1 vaccine which expresses six antigens of gp160, gag, pol, rev, tat and nef of HIV subtype B'/C. A replicating DNA vaccine and a non-replicating recombinant vaccinia virus vector, both expressing the six antigens mentioned above, were used to immune BALB/c (H-2d) mice in a prime-boost regiment. The six peptide libraries of HIV B'/C corresponding respectively to the six complete antigens were pooled according to a designed matrix format and used to test for IFN-gamma production from splenocytes of immunized mice by an enzyme-linked immunospot (IFN-gamma ELISPOT) assay. The ELISPOT data indicated that two of seven Gag-specific T-cell epitope peptides were identified to be the novel epitopes. One of three Pol-specific T-cell epitope is unreported. One novel epitope was confirmed in two gp160-specific T-cell epitope peptides. One Nef-specific T-cell epitope was identified. Three Tat-specific T-cell epitope peptides were continuous sequences in Tat peptide library and all contained either complete or partial sequence reported. Rev-specific T-cell epitope was not be found. The specific T-cell epitopes (H-2d restricted) were identified by IFN-7 ELISPOT assay, which could be used to detect the cellular immune response of BALB/c mice immunized with the HIV-1 vaccine expressing six antigens of gp160, gag, pol, rev, tat and nef of HIV subtype B'/C.


Assuntos
Animais , Feminino , Humanos , Camundongos , Sequência de Aminoácidos , ELISPOT , Métodos , Epitopos de Linfócito T , Química , Genética , Alergia e Imunologia , Antígenos H-2 , Química , Genética , Alergia e Imunologia , Antígenos HIV , Química , Genética , Alergia e Imunologia , Infecções por HIV , Alergia e Imunologia , Virologia , HIV-1 , Classificação , Genética , Alergia e Imunologia , Antígeno de Histocompatibilidade H-2D , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Mapeamento de Peptídeos , Métodos
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...