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1.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-636511

RESUMO

5-aminosalicylic acid (5-ASA) is drug of choice for the treatment of ulcerative colitis (UC). In this study, the efficacy of topical versus oral 5-ASA for the treatment of UC was examined as well as the action mechanism of this medication. A flexible tube was inserted into the rat cecum to establish a topical administration model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced UC. A total of 60 rats were divided into sham operation group (receiving an enema of 0.9% saline solution instead of the TNBS solution via the tube), model group, topical 5-ASA group, oral Etiasa group (a release agent of mesalazine used as positive control) and oral 5-ASA group (n=12 each). Different treatments were administered 1 day after UC induction. The normal saline (2 mL) was instilled twice a day through the tube in the sham operation group and model group. 5-ASA was given via the tube in the topical 5-ASA group (7.5 g/L, twice per day, 100 mg/kg), and rats in the oral Etiasa group and oral 5-ASA group intragastrically received Etiasa (7.5 g/L, twice per day, 100 mg/kg) and 5-ASA (7.5 g/L, twice per day, 100 mg/kg), respectively. The body weight was recorded every day. After 7 days of treatment, blood samples were drawn from the heart to harvest the sera. Colonic tissues were separated and prepared for pathological and related molecular biological examinations. The concentrations of 5-ASA were detected at different time points in the colonic tissues, feces and sera in different groups by using the high pressure liquid chromatography (HPLC). The results showed that the symptoms of acute UC, including bloody diarrhea and weight loss, were significantly improved in topical 5-ASA-treated rats. The colonic mucosal damage, both macroscopical and histological, was significantly relieved and the myeloperoxidase activity was markedly decreased in rats topically treated with 5-ASA compared with those treated with oral 5-ASA or Etiasa. The mRNA and protein expression of IL-1β, IL-6, and TNF-α was down-regulated in the colonic tissue of rats topically treated with 5-ASA, significantly lower than those from rats treated with oral 5-ASA or Etiasa. The concentrations of 5-ASA in the colonic tissue were significantly higher in the topical 5-ASA group than in the oral 5-ASA and oral Etiasa groups. It was concluded that the topical administration of 5-ASA can effectively increase the concentration of 5-ASA in the colonic tissue, decrease the expression of proinflammatory cytokines, alleviate the colonic pathological damage and improve the symptoms of TNBS-induced acute UC in rats.

2.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-321348

RESUMO

This paper aims to summarize the achievements during the implementation process of good agricultural practice (GAP) in Chinese Materia Medica (CMM), and on basis of analyzing the existing problems of GAP, to propose further implementation of GAP in TCM growing. Since the launch of GAP in CMM growing ten years ago, it has acquired great achievements, including: (1) The promulgation of a series of measures for the administration of the GAP approval in the CMM growing; (2) The expanded planting area of CMM; (3) The increased awareness of standardized CMM growing among farmers and enterprises; (4) The establishment of GAP implementation bases for CMM growing; (5) The improvement of theory and methodology for CMM growing; (6) The development of a large group of experts and scholars in GAP approval for CMM production. The problems existing in the production include: (1) A deep understanding of GAP and its certification is still needed; (2) The distribution of the certification base is not reasonable; (3) The geo-economics effect and the backward farming practices are thought to be the bottlenecks in the standardization of CMM growing and the scale production of CMM; (4) Low comparative effectiveness limits the development of the GAP; (5) The base of breeding improved variety is blank; (6) The immature of the cultivation technique lead to the risk of production process; (7) The degradation of soil microbial and the continuous cropping obstacle restrict the sustainable development of the GAP base. To further promote the health and orderly GAP in the CMM growing, the authors propose: (1) To change the mode of production; (2) To establish a sound standard system so as to ensure quality products for fair prices; (3) To fully consider the geo-economic culture and vigorously promote the definite cultivating of traditional Chinese medicinal materials; (4) To strengthen the transformation and generalization of basic researches and achievements, in order to provide technical support for the CMM production; (5) To deepen the understanding of GAP, to vigorously promote ecological planting and precision agriculture, in order to overcome the continuous cropping obstacle. The authors think that despite the fact that we are still facing with a huge array of management and technological problems, the GAP in the CMM growing has already enjoyed widespread support and showed great potential. In the future, with people's deeper understanding of GAP and the great progress of the science and technology, the GAP will constantly be fused with the theory, methodology and technology in the modern agriculture like precision agriculture, eco-agriculture and etc.


Assuntos
Humanos , Agricultura , Economia , Métodos , Padrões de Referência , China , Medicamentos de Ervas Chinesas , Economia , Padrões de Referência , Materia Medica , Química , Economia , Padrões de Referência , Plantas Medicinais , Química
3.
Artigo em Inglês | WPRIM (Pacífico Ocidental) | ID: wpr-251360

RESUMO

5-aminosalicylic acid (5-ASA) is drug of choice for the treatment of ulcerative colitis (UC). In this study, the efficacy of topical versus oral 5-ASA for the treatment of UC was examined as well as the action mechanism of this medication. A flexible tube was inserted into the rat cecum to establish a topical administration model of 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced UC. A total of 60 rats were divided into sham operation group (receiving an enema of 0.9% saline solution instead of the TNBS solution via the tube), model group, topical 5-ASA group, oral Etiasa group (a release agent of mesalazine used as positive control) and oral 5-ASA group (n=12 each). Different treatments were administered 1 day after UC induction. The normal saline (2 mL) was instilled twice a day through the tube in the sham operation group and model group. 5-ASA was given via the tube in the topical 5-ASA group (7.5 g/L, twice per day, 100 mg/kg), and rats in the oral Etiasa group and oral 5-ASA group intragastrically received Etiasa (7.5 g/L, twice per day, 100 mg/kg) and 5-ASA (7.5 g/L, twice per day, 100 mg/kg), respectively. The body weight was recorded every day. After 7 days of treatment, blood samples were drawn from the heart to harvest the sera. Colonic tissues were separated and prepared for pathological and related molecular biological examinations. The concentrations of 5-ASA were detected at different time points in the colonic tissues, feces and sera in different groups by using the high pressure liquid chromatography (HPLC). The results showed that the symptoms of acute UC, including bloody diarrhea and weight loss, were significantly improved in topical 5-ASA-treated rats. The colonic mucosal damage, both macroscopical and histological, was significantly relieved and the myeloperoxidase activity was markedly decreased in rats topically treated with 5-ASA compared with those treated with oral 5-ASA or Etiasa. The mRNA and protein expression of IL-1β, IL-6, and TNF-α was down-regulated in the colonic tissue of rats topically treated with 5-ASA, significantly lower than those from rats treated with oral 5-ASA or Etiasa. The concentrations of 5-ASA in the colonic tissue were significantly higher in the topical 5-ASA group than in the oral 5-ASA and oral Etiasa groups. It was concluded that the topical administration of 5-ASA can effectively increase the concentration of 5-ASA in the colonic tissue, decrease the expression of proinflammatory cytokines, alleviate the colonic pathological damage and improve the symptoms of TNBS-induced acute UC in rats.


Assuntos
Animais , Masculino , Ratos , Administração Oral , Administração Tópica , Anti-Inflamatórios não Esteroides , Farmacologia , Colite Ulcerativa , Tratamento Farmacológico , Colo , Metabolismo , Patologia , Regulação para Baixo , Esquema de Medicação , Expressão Gênica , Imuno-Histoquímica , Interleucina-1beta , Genética , Metabolismo , Interleucina-6 , Genética , Metabolismo , Mucosa Intestinal , Metabolismo , Patologia , Mesalamina , Farmacologia , Peroxidase , Metabolismo , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , Resultado do Tratamento , Ácido Trinitrobenzenossulfônico , Fator de Necrose Tumoral alfa , Genética , Metabolismo
4.
Chinese Journal of Neuromedicine ; (12): 466-469, 2013.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1033768

RESUMO

Objective To explore the effect of Yindan Xinnaotong capsules (YDXNT) on the reactive nitrogen intermediates in the cerebral cortex and learning and memory abilities of rats with cerebral ischemia.Methods Thirty rats were randomly divided into sham-operated group,vehicle group and Yindan Xinnaotong capsule treatment group.The animal models of cerebral ischemia in the later two groups were established by two vessel occlusion.Yindan Xinnaotong capsule 60 mg/(kg·d) via inttragastric administration was given to the Yindan Xinnaotong capsule treatment group for a consecutive four weeks.The learning and memory ability changes were tested with Morris water maze and the contents of nitric oxide (NO) and inducible nitric-oxide synthase (iNOS) in the cerebral cortex were measured by colorimetric method.Results As compared with the vehicle group,the Yindan Xirnaotong capsule treatment group and sham-operated group had significantly shorter escape latency and longer searching time in the quadrant of primary platform (P<0.05); As compared with the vehicle group,the Yindan Xinnaotong capsule treatment group had significant decreased contents of NO and iNOS in the cerebral cortex (P<0.05).Conclusion Yindan Xinnaotong capsules can improve the learning and memory abilities of rats with cerebral ischemia by lowering the activity of NO and iNOS in the cerebral cortex.

5.
Chinese Journal of Oncology ; (12): 249-255, 2013.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-284197

RESUMO

<p><b>OBJECTIVE</b>To investigate the effects of a proliferation-inducing ligand (APRlL) on colorectal cancer (CRC) cell growth and migration, and to observe the role of APRIL in CRC biological behavior.</p><p><b>METHODS</b>The siRNA plasmid vector targeting APRIL gene (APRIL-siRNA) was transfected into human colorectal cancer SW480 cells and recombinant human APRIL (rhAPRIL) was used to stimulate human colorectal cancer HCT-116 cells. Cell proliferation activity was analyzed using cell counting kit-8 (CCK-8), cell cycle was detected by flow cytometry, and the protein expression of cyclin D1, p21 and Bcl-2 was detected by Western blot analysis. Tumor cell migration and invasion were measured by Transwell chambers. RT-PCR was applied to examine the mRNA expression level of MMP-2 and MMP-9. APRIL-siRNA was used to transfect directly SW480 cells, which were injected subcutaneously into nude mice, then the tumor growth and metastasis were observed.</p><p><b>RESULTS</b>Cell proliferation ability of APRIL-siRNA-transfected SW480 cells was drastically repressed, and the percentage of G0/G1 phase cells was significantly increased (t = 4.12, P < 0.05), accompanied with depressed cyclin D1, Bcl-2 expression and elevated p21 expression. Cell proliferation ability of rhAPRIL-stimulated HCT-116 cells was promoted with a decreased G0/G1 phase ratio (t = 3.31, P < 0.05). cyclin D1 and Bcl-2 protein expression was up-regulated while p21 was down-regulated by rhAPRIL stimulation. Metastatic and invasive capacities of APRIL-siRNA-transfected SW480 cells were significantly inhibited compared with their respective controls (both P < 0.05), accompanied with the deregulated MMP-2 and MMP-9 mRNA expression. Metastatic and invasive capacities of rhAPRIL-stimulated HCT-116 cells were promoted with up-regulated MMP-2 and MMP-9 mRNA expression(both P < 0.05). Tumor growth in the group transfected with APRIL-siRNA appeared to be slower than that in the control groups and the expression of MMP-2, MMP-9 in tumor tissues was depressed in the APRIL-siRNA group.</p><p><b>CONCLUSIONS</b>APRIL facilitates tumor growth and metastasis, and is associated with carcinogenesis and prognosis. Our findings suggest that APRIL might be used as a novel target for the intervention and therapy of colorectal cancer.</p>


Assuntos
Animais , Feminino , Humanos , Camundongos , Ciclo Celular , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Neoplasias Colorretais , Metabolismo , Patologia , Ciclina D1 , Metabolismo , Vetores Genéticos , Células HCT116 , Células HT29 , Metaloproteinase 2 da Matriz , Metabolismo , Metaloproteinase 9 da Matriz , Metabolismo , Camundongos Nus , Invasividade Neoplásica , Metástase Neoplásica , Transplante de Neoplasias , Plasmídeos , Proteínas Proto-Oncogênicas c-bcl-2 , Metabolismo , Proteínas Proto-Oncogênicas p21(ras) , Metabolismo , RNA Mensageiro , Metabolismo , RNA Interferente Pequeno , Genética , Transfecção , Carga Tumoral , Membro 13 da Superfamília de Ligantes de Fatores de Necrose Tumoral , Genética , Metabolismo
6.
Chinese Journal of Virology ; (6): 178-184, 2012.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-354751

RESUMO

The transmembrane protein (TM) encoded by gp37 gene plays a critical role when virus fusion with cell membrane occurs. Several highly conserved regions in TM are important targets for antivirus studies. Studies on structure and function of TM will provide basic information for anti-retrovirus, especially for avian leukosis virus. In the study, gp37 gene was amplified by PCR from the Chinese strain ALV-J-WS0701. The gp37 gene was cloned into pMD18-T vector, and was sequenced. Then, pFast-BacHTb-gp37 vector was constructed and expressed by baculovirus expression vector system. The expression product of gp37 gene was analyzed by indirect immunofluorescence assay and Western blot. The results showed that positive green fluorescence was present in sf9 cells infected with recombinant virus and a protein band with a molecular weight of 21kD was present in Western blot. It is concluded that gp37 gene was expressed in sf9 cells infected with recombinant virus successfully.


Assuntos
Animais , Leucose Aviária , Virologia , Vírus da Leucose Aviária , Classificação , Genética , Linhagem Celular , Galinhas , Clonagem Molecular , Expressão Gênica , Spodoptera , Proteínas do Envelope Viral , Genética , Metabolismo
7.
Chinese Journal of Neuromedicine ; (12): 369-371, 2012.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1033511

RESUMO

Objective To investigate the effect ofYindanxinnaotong (YDXNT) capsule on learning and memory abilities, super oxide dismutase (SOD) activity and malondialdehyde (MDA) content in the hippocampus of rat models of vascular dementia (VD). Methods Animals were randomly divided into sham-operated group,VD vehicle group (Group VD) and YDXNT treatment group (Group VD+YDXNT); the 4-vessel occlusion was employed to establish the VD rat models in the Group VD and Group VD+YDXNT,while rats in the sham-operated group were not performed occlusion of the bilateral common carotid artery.Rats of the Group VD+YDXNT were given 1 mL liquid (9 granules of YDXNT into 360 mL normal saline) through intragastric administration 3 times daily for a consecutive 4 weeks; and rats of the other 2 groups were given the same volume of saline.The changes of learning and memory abilities were observed by Morris water maze test; the activity of SOD and the content of MDA in the hippocampus were measured. Results As compared with those in rats of the Group VD, the learning and memory abilities in rats of the Group VD+YDXNT were significantly improved (P<0.05):time in finding the platform was obviously decreased and swimming distance in finding the platform was obviously shortened in rats of Group VD+YDXNT.As compared with those in rats of the Group VD,the activity of SOD in the hippocampus was significantly increased and the content of MDA was obviously decreased in rats of the Group VD+YDXNT (P<0.05). Conclusion YDXNT could increase oxygen radical metabolism to improve the learning and memory abilities of rats with VD.

8.
Chinese Journal of Neuromedicine ; (12): 590-593, 2012.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1033552

RESUMO

Objective To study the effect of Yindan Xinnaotong Capsules (YDXNT) on impairment of learning and memory in rats with vascular dementia (VD) and acetylcholinesterase (AchE) level of the hippocampus,and observe the mechanism of YDXNT in VD. Methods Bilateral carotid arteries were repeatedly blocked and performed reperfusion with nitroprusside sodium by intraperitoneal injection to establish the rat models of VD.VD rat models were randomly divided into VD model group (n=10) and YDXNT treatment group (n=10),and another 10 condition-matched rats were selected as the sham-operated group.At 30 min after regaining consciousness and before execution,the neurological scale was performed on these rats; the effect of YDXNT on learning and memory in rats were tested by step-down avoidance tests; rats' brains were dissected out on the ice for detecting the activity of AchE.Results The scores of neurological scale in the YDXNT treatment group and VD model group were obviously higher than those in the sham-operated group (P<0.05); the scores of neurological scale in the YDXNT treatment group were significantly lower than those in the VD model group (P<0.05).In the step-down avoidance tests,the rats in the YDXNT treatment group had significantly shorter reaction time,less number of errors and longer latency than rats in the VD model group (P<0.05).As compared with that in the VD model group,the activity of AChE in the hippocampus of YDXNT treatment group was significantly decreased (P<0.05). Conclnsion YDXNT can ameliorate the learning and memory deficits of VD model rats and the mechanism may be its involvement in inhibiting the AChE activity.

9.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-321230

RESUMO

<p><b>OBJECTIVE</b>To explore the effects of postoperative enteral immunonutrition on inflammatory response and immunologic function in patients with gastrointestinal tumor.</p><p><b>METHODS</b>Clinical data of 106 gastrointestinal malignant tumor patients with malnutrition who were treated in the Department of General Surgery, the People's Hospital of Cangzhou in Hebei province from January 2008 to June 2010 were prospectively collected. Patients were randomized into two groups, including enteral immunonutrition group(n=53) and common enteral nutrition group(n=53). Related immunological indices and C-reaction protein were measured on preoperative day 5 and postoperative day 1 and 9.</p><p><b>RESULTS</b>The general information and preoperative immunological indices were comparable between the two groups(P>0.05). On postoperative day 9, levels of CD4, CD4/CD8, IgG, lymphocyte, NK cells, and complement C3, C4, and CH50 in the enteral immunonutrition group were higher than those in common enteral nutrition group. Serum C-reaction protein level was lower than that in control group, and the difference was statistically significant (P<0.05). Postoperative infection rate was 3.8%(2/53) in enteral immunonutrition group, significantly lower than that in control group with an infection rate of 15.1%(8/53)(P<0.05). The mean postoperative hospital stay of the two groups were (8.1±1.1) d and (9.2±2.1) d, respectively, and the difference was statistically significant(P<0.05).</p><p><b>CONCLUSION</b>For gastrointestinal malignant tumor patients with malnutrition, the use of enteral immunonutrition can alleviate the postoperative trauma and inflammatory response, improve the immune function, thus can reduce the occurrence of postoperative infection, and accelerate patient recovery.</p>


Assuntos
Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem , Proteína C-Reativa , Relação CD4-CD8 , Proteínas do Sistema Complemento , Alergia e Imunologia , Nutrição Enteral , Neoplasias Gastrointestinais , Alergia e Imunologia , Terapêutica , Inflamação , Alergia e Imunologia , Células Matadoras Naturais , Alergia e Imunologia , Desnutrição , Terapêutica , Período Pós-Operatório , Estudos Prospectivos , Resultado do Tratamento
10.
Chinese Journal of Neuromedicine ; (12): 847-849, 2010.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-1033073

RESUMO

Objective To investigate the effects of Yindanxinnaotong capsule on hemorrheology indexes and blood lipid levels in elderly patients with lacunar infarction (LI).Methods The hemorrheology indexes and blood lipid levels were measured in 62 elderly patients with LI, and then,compared with those of 50 healthy subjects. The hemorrheology indexes and lipid levels were re-measured to assess the therapeutic effect of Yindanxinnaotong capsule in 50 patients 45 d after the treatment. Results The hemorrheology indexes and the levels of total cholesterol, triglyceride, and low-density lipoprotein cholesterol in patient group were significantly higher than those in healthy control group before the treatment (P<0.05). The hemorrheology indexes and blood lipid levels after the treatment with Yindanxinnaotong capsule in patient group were significantly improved as compared with those before treatment (P<0.05). Conclusion The obvious abnormalities of hemorrheology indexes and blood lipid levels are found in aged patients with LI and Yindanxinnaotong capsule can improve these abnormalities.

11.
Chinese Journal of Pathology ; (12): 118-121, 2007.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-333956

RESUMO

<p><b>OBJECTIVE</b>To study the clinicopathologic features of sclerosing angiomatoid nodular transformation of spleen and its differential diagnosis.</p><p><b>METHODS</b>The clinicopathologic characteristics and immunophenotype of 4 cases of sclerosing angiomatoid nodular transformation of spleen were studied.</p><p><b>RESULTS</b>Histologically, all cases were characterized by multiple angiomatoid nodules of various sizes in a fibrosclerotic stroma. The nodules were round and sometimes convoluted. They were composed of slit-like, irregular-shaped or slightly dilated vascular spaces lined by plump endothelial cells and interspersed with a population of spindly or ovoid cells. Immunohistochemical study showed a heterogeneous staining pattern, with the lining cells of the small capillaries expressing CD34 and those of the sinusoid-like structures expressing CD8. CD31 highlighted both the lining cells and interspersed cells, resulting in a complex meshwork. The lining cells were also focally positive for CD68. Smooth muscle actin revealed conglomerates of spindly shaped cells around and between the vascular channels. These spindly shaped cells in the intervening stroma were focally positive for actin, but negative for desmin, CD21 and CD35.</p><p><b>CONCLUSIONS</b>Sclerosing angiomatoid nodular transformation is a rarely encountered benign lesion of the spleen, which should be distinguished from other angiomatoid tumors and tumor-like lesions.</p>


Assuntos
Adulto , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Angiomatose , Metabolismo , Patologia , Cirurgia Geral , Antígenos CD34 , Metabolismo , Antígenos CD8 , Metabolismo , Diagnóstico Diferencial , Seguimentos , Hamartoma , Patologia , Hemangioma , Patologia , Imuno-Histoquímica , Molécula-1 de Adesão Celular Endotelial a Plaquetas , Metabolismo , Esclerose , Patologia , Baço , Patologia , Esplenectomia , Esplenopatias , Metabolismo , Patologia , Cirurgia Geral , Neoplasias Esplênicas , Patologia
12.
Chinese Journal of Biotechnology ; (12): 315-319, 2005.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-305278

RESUMO

In order to differently diagnose avian influenza virus (AIV) subtypes, the HA gene of AIV H9 subtype was cloned, expressed and utilized in an enzyme-linked immunoad sorbent assay (ELISA). HA gene (1683bp) of H9N2 AIV was amplified by RT-PCR from a strain of field isolated H9N2 AIV, and its identity was confirmed by sequencing. The HA gene was subcloned into prokaryotic expression vector pGEX-KG with its secretion signal sequence removed. The expressed HA-GST fusion protein in E. coli BL21 was characterized by SDS-PAGE and western blotting analysis as a 90kD protein with immunogenicity. The fusion protein was present primarily in inclusion bodies and was purified via denaturation and renenaturation. The HA-GST fusion protein was used to establish an indirect ELISA for the detection of antibodies to H9 subtypes of AIV. The assay has 91.57% specificity to H9 AIV, 92.31% sensitivity and excellent reduplication. It could be used to differently detect antibodies to H9 AIV.


Assuntos
Humanos , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática , Escherichia coli , Genética , Metabolismo , Glicoproteínas de Hemaglutininação de Vírus da Influenza , Genética , Vírus da Influenza A Subtipo H9N2 , Genética , Influenza Humana , Diagnóstico , Virologia , Proteínas Recombinantes , Genética
13.
Artigo em Chinês | WPRIM (Pacífico Ocidental) | ID: wpr-685680

RESUMO

Objective To develop a convenient method efficiently expands the frequency of specific CTLS.Methods We used different concentrations of CMV-speeific epitope peptides pp65 to stimulate PBMCs for expansion of CMV-specific CTLs.CMV-specifie CTLs were doubly labeled by tetramers-PE and CD_8-FITC for FACS analysis.Results The method expands CMV-speeific CTLs efficiently.CMV-specific CTLs were expanded from 1% to 20% of PBMCs quickly(namely 40% of CD_8~+ T cells).The method provided a large number of cells with tetramer staining of CD_8~+ T cells for FACS analysis from a single blood sampling.Conclusions Peptides stimulation methods are convenient,easy to operate and expanded CMV- specific CTLs efficiently.The increased frequencies of CMV-specific CTLs allowed the data of different individuals to be easily compared and sequentially evaluated.The methods lay the base for adoptive immunotherapy to prevent CMV disease.

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