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1.
Zhonghua Yi Xue Za Zhi ; 100(18): 1376-1379, 2020 May 12.
Artigo em Zh | MEDLINE | ID: mdl-32392986

RESUMO

Objective: To evaluate the efficacy of coronary artery bypass grafting (CABG) surgery in left ventricular dysfunction patients complicated with different degrees of ischemic mitral regurgitation (IMR). Methods: The clinical data of 525 patients (428 males and 97 females) undergoing CABG in Fuwai Hospital Chinese Academy of Medical Sciences Shenzhen, and Tianjin Medical University General Hospital between January 2015 and December 2018 were collected. The average age was (61±7) years old. Among them, the patients with moderate to serve IMR and left ventricular ejection fraction(LVEF)≤40% were further selected, and the outcomes of CABG were analyzed. Results: In total, 67 patients (48 males and 19 females) with moderate to severe IMR and LVEF≤40% were enrolled, among which 52 patients had moderate IMR, with a LVEF of 38%(35%, 40%). Transesophageal echocardiography (TEE) of 52 cases displayed no damage of papillary muscles, and ventricular wall motion was improved after CABG. Therefore, no treatment on the mitral valve was performed in this group. Six patients were with moderate-severe mitral insufficiency, with a LVEF of 38%(35%, 39%). After surgery, TEE found that the ventricular wall motion and regurgitation were improved, and the mitral valve structures were well. Thus, mitral valves were not treated in these patients. Nine patients were with severe mitral regurgitation, with a LVEF of 38%(35%, 39%). Two of them received valve repair because the papillary muscle function and the ring were well. Another 7 patients received valve replacements because the valve ring was dilatated and the leaflet was prolapsed. All patients recovered well. The LVEF increased significantly at 6 months after surgery [47%(45%, 48%) vs 38%(35%, 39%), P=0.024], and the left ventricular end diastolic diameter also became smaller [57(56, 59) mm vs 61(59, 64) mm, P=0.002]. Conclusions: For patients suffered from left ventricular dysfunction complicated with IMR, TEE is crucial to evaluate the valve function. To those with moderate-severe regurgitation, if papillary muscle function and the ring were seriously affected by ischemia, the valve replacement could facilitate the improvement of postoperative cardiac function.


Assuntos
Insuficiência da Valva Mitral , Isquemia Miocárdica , Disfunção Ventricular Esquerda , Idoso , Ponte de Artéria Coronária , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência da Valva Mitral/cirurgia , Isquemia Miocárdica/cirurgia , Volume Sistólico , Resultado do Tratamento , Função Ventricular Esquerda
2.
Zhonghua Yu Fang Yi Xue Za Zhi ; 52(10): 1039-1044, 2018 Oct 06.
Artigo em Zh | MEDLINE | ID: mdl-30392324

RESUMO

Objective: To establish a triple-color pseudovirion-based neutralization assay (PBNA) and evaluate its capability of detecting immunogenicity of the sera generated by the immunization of HPV 9-valent vaccine. Methods: HPV pseudovirus (PsVs) 6/11/16/18/31/33/45/52/58 with the encapsidated fluorescence expressing red fluorescent plasmid N31-MCHREEY, green fluorescent N31-EGFP or blue fluorescent N31-mTagBFP were generated. The concentration of HPV PsVs and the infection titers of HPV PsVs were detected by double-antibody sandwich ELISA and TCID(50), respectively. The single- and triple color HPV 16/33/45 PsVs were used to detect the neutralization titers of mice sera immunized with HPV 9-valent vaccine and confirmed the accuracy and specificity of the triple-color PBNAs. Then, the single- and triple color HPV 6/11/18/31/33/45/52/58 PsVs were employed to detect the neutralization titers of cynomolgus macaques sera immunized with HPV 9-valent vaccine and determined whether the triple-color PBNAs could be applied to evaluate the immunogenicity of the sera generated by the immunization of HPV9-valent vaccine. Results: The concentration of HPV16 PsVs encapsulating green, red or blue fluorescent plasmid was 5.0 to 6.0 µg/ml and HPV6/11/18/31/33/45/52/59 triple-color HPV PsVs was about 1.0 to 3.0 µg/ml. 9 types HPV PsVs containing EGFP, Mcherry or mTagBFP reporter plasmid were obtained and the concentration can meet the need of neutralization detection. 9 types single-color fluorescent HPV PsVs had similar infectivity against 293FT cells with the infection titer values between 1×10(4) and 1×10(5). The results of PBNAs showed that there was no significant difference in the anti-HPV neutralization titers of mice sera induced by HPV 9-valent vaccine between single-color and triple-color HPV16/33/45 PsVs (P>0.05). Similarly, there was also no significant difference in the anti-HPV neutralization titers of cynomolgus macaques sera induced by HPV 9-valent vaccine between single-color and triple-color HPV6/11/18/31/33/45/52/58 PsVs (P>0.05). Conclusion: We successfully established the triple-color PBNAs and verified the accuracy and specificity of triple-color PBNAs consistent with single-color PBNAs. The triple-color PBNAs can be applied to evaluate the immunogenicity of HPV 9-valent vaccine's immune serum.


Assuntos
Fluorimunoensaio/métodos , Testes de Neutralização/métodos , Papillomaviridae/imunologia , Animais , Anticorpos Antivirais/sangue , Cor , Papillomavirus Humano 16/isolamento & purificação , Humanos , Camundongos , Infecções por Papillomavirus , Vacinas contra Papillomavirus/imunologia , Reprodutibilidade dos Testes
3.
J Theor Biol ; 320: 86-99, 2013 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-23261980

RESUMO

Antiangiogenic drugs were developed with the aim to inhibit the formation of intratumoral blood vessels and in consequence the growth of solid tumors. As these drugs are generally combined with classical cytotoxic drugs in the treatment of cancer patients, finding the optimal combinations remains a complex challenge due to possible interactions of the antiangiogenic compound with the hemodynamic property of the treated tumor. To analyze this problem, we developed a multi-scale model of vascular tumor growth combining a molecular model of VEGF signaling pathways and a tissue model of the tumor expansion including the dynamics of cellular and tissue processes of tumor growth and response to treatments. We addressed the potential impact of antiangiogenic drug by defining a new index of vasculature quality which depends on the balance between stable and unstable vessels within the tumor mass. Our goal was to investigate the interactions between a chemotherapy and a antiangiogenic treatment, and, by simulating the model, to identify the optimal delay of chemotherapy delivery after the administration of the antiangiogenic compound. This theoretical analysis could be used in the future to optimize antiangiogenic drug delivery in preclinical settings and to facilitate the translation from preclinical to clinical studies.


Assuntos
Inibidores da Angiogênese/farmacologia , Modelos Biológicos , Neoplasias Experimentais/tratamento farmacológico , Neovascularização Patológica/tratamento farmacológico , Animais , Citotoxinas/farmacologia , Humanos , Camundongos , Proteínas de Neoplasias/metabolismo , Neoplasias Experimentais/patologia , Neoplasias Experimentais/fisiopatologia , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Neovascularização Patológica/fisiopatologia , Transdução de Sinais/efeitos dos fármacos , Fator A de Crescimento do Endotélio Vascular/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto/métodos
4.
Cancer Res ; 54(7): 1843-8, 1994 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-8137299

RESUMO

We previously showed that the Epstein-Barr virus, which encodes the BARF1 gene, could transform rodent fibroblasts. In this work, the expression of the BARF1 gene was studied in the human Louckes B-lymphocyte cell line. Introduction of the BARF1 open reading frame under the control of the Mo-MuLV LTR promotor into nontumorigenic Louckes lymphoid cells led to the activation of the c-myc protooncogene and increased expression of the B-cell surface proteins, the transferrin receptor, CD21, and CD23. BARF1-expressing cells induced a diffuse lymphoma-like tumor in newborn rats treated with anti-thymocyte serum that was, however, transient and regressed after 3-4 weeks as the immune system recovered. The tumor induction was similar to that observed with lymphoid cell lines in vitro generated by infection with the B95-8 virus strain, in which lytic antigens are expressed at low levels. After long-term culture, Louckes cell clones lost expression of the BARF1 gene and were unable to induce tumors.


Assuntos
Linfócitos B/metabolismo , Regulação Viral da Expressão Gênica , Genes Virais , Genes myc , Herpesvirus Humano 4/metabolismo , Proteínas Virais/biossíntese , Antígenos CD/biossíntese , Linfoma de Burkitt , Linhagem Celular , Células Clonais , Imunofluorescência , Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Herpesvirus Humano 4/genética , Humanos , Vírus da Leucemia Murina de Moloney/genética , Fases de Leitura Aberta , Regiões Promotoras Genéticas , Receptores de Complemento 3d/biossíntese , Receptores de IgE/biossíntese , Receptores da Transferrina/biossíntese , Células Tumorais Cultivadas , Proteínas Virais/genética
5.
Cancer Res ; 53(13): 3125-8, 1993 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-8319220

RESUMO

The ability of interleukin-4 (IL-4) to mediate an antitumor response to human gliomas was studied in vivo in nude mice. To allow the effect of IL-4 to be exerted over a relatively short distance and at an optimal concentration, a transfected tumor cell line expressing a high level of IL-4 was used in mixed tumor transplantation assays. There was a significant inhibition of growth of the U87 human glioma line when the IL-4-secreting cell line, LT-1, was implanted s.c. with the glioma in 5 nude mice when compared to contralateral control tumors consisting of the U87 glioma and IL-4-negative control cells. In addition, there was a prolongation of survival when U87 along with IL-4-secreting cells were implanted intracerebrally in 12 nude mice compared to 12 control nude mice implanted with U87 and IL-4-negative control cells and 11 control animals receiving U87 alone. Histological analysis 4 days after i.c. inoculation revealed the presence of a dramatic eosinophil infiltrate and tumor necrosis. The absence of viable glioma cells as well as resolution of inflammation 19 days after treatment suggests the potential for complete tumor regression without ongoing inflammatory sequelae resulting from cytokine treatment.


Assuntos
Neoplasias Encefálicas/terapia , Glioma/terapia , Imunoterapia , Interleucina-4/metabolismo , Plasmocitoma/metabolismo , Adjuvantes Imunológicos/farmacologia , Animais , Feminino , Humanos , Injeções Intraventriculares , Injeções Subcutâneas , Interleucina-4/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Transplante de Neoplasias , Plasmocitoma/patologia , Ratos , Proteínas Recombinantes/metabolismo , Proteínas Recombinantes/farmacologia
6.
Cancer Res ; 54(22): 5745-51, 1994 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-7954393

RESUMO

Survival of rats harboring cerebral 9L gliosarcomas can be significantly extended by an intratumoral inoculation with a herpes simplex virus vector, designated as hrR3. This vector, which bears the lacZ reporter gene, is defective in the gene encoding ribonucleotide reductase, allowing for replication in dividing tumor cells but not in postmitotic neural cells. It also possesses an intact viral thymidine kinase (TK) gene, which confers chemosensitivity to ganciclovir. In this study, the ability of ganciclovir to potentiate the antitumor effect of hrR3 was evaluated. In culture, there was a 23% decrease in the growth of 9L cells treated with hrR3 plus ganciclovir compared to hrR3 alone (P < 0.01). The combination of hrR3 plus ganciclovir led to the long-term survival of 48% of rats harboring intracerebral 9L gliosarcomas compared to 20% survival in the hrR3 group (P < 0.05). Ganciclovir treatment had no effect on the growth of tumor cells in vitro or in vivo when a herpes simplex virus vector with a defective TK gene was used. Immunocytochemistry confirmed selective expression of the TK gene in cells within the tumor. These findings indicate that the TK gene can potentiate the antitumor effect of the hrR3 herpes simplex virus vector and provide the basis for placing additional therapeutic genes in the genome of hrR3.


Assuntos
Neoplasias Encefálicas/terapia , Ganciclovir/uso terapêutico , Terapia Genética/métodos , Gliossarcoma/terapia , Simplexvirus/genética , Timidina Quinase/genética , Animais , Neoplasias Encefálicas/enzimologia , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/mortalidade , Neoplasias Encefálicas/patologia , Terapia Combinada , Vetores Genéticos/genética , Gliossarcoma/enzimologia , Gliossarcoma/genética , Gliossarcoma/mortalidade , Gliossarcoma/patologia , Masculino , Ratos , Ratos Endogâmicos F344 , Simplexvirus/enzimologia , Timidina Quinase/análise , Células Tumorais Cultivadas
7.
Cancer Res ; 58(16): 3529-32, 1998 Aug 15.
Artigo em Inglês | MEDLINE | ID: mdl-9721854

RESUMO

We examined the efficacy of suicide gene therapy for nitrosomethylurea-induced mammary tumors in rats. Individual tumors were directly injected with a retrovirus-producing cell line that releases retroviral vectors that transduce the herpes simplex virus type 1 thymidine kinase (HSV1-TK) gene. HSV1-TK specifically converts the nucleoside analogue ganciclovir (GCV) into a toxic metabolite. Compared to control rats receiving saline, we observed a significant tumor regression of the injected tumors following GCV administration, accompanied by a stromal inflammation and an extensive lymphocyte infiltration invading the tumor epithelium. It is noteworthy that the neighboring uninjected tumors also regressed, demonstrating the occurrence of a distant bystander effect. This is the first demonstration that HSV1-TK/GCV can efficiently treat multiple solid tumors directly generated from an epithelial tissue.


Assuntos
Antimetabólitos/uso terapêutico , Ganciclovir/uso terapêutico , Terapia Genética/métodos , Herpesvirus Humano 1/enzimologia , Neoplasias Mamárias Experimentais/terapia , Timidina Quinase/uso terapêutico , Animais , Antimetabólitos/metabolismo , Carcinógenos , Feminino , Ganciclovir/metabolismo , Herpesvirus Humano 1/genética , Neoplasias Mamárias Experimentais/induzido quimicamente , Neoplasias Mamárias Experimentais/patologia , Metilnitrosoureia , Neoplasias Primárias Múltiplas/induzido quimicamente , Neoplasias Primárias Múltiplas/patologia , Neoplasias Primárias Múltiplas/terapia , Ratos , Ratos Sprague-Dawley , Timidina Quinase/genética , Timidina Quinase/metabolismo , Transfecção
8.
Oncogene ; 14(25): 3073-81, 1997 Jun 26.
Artigo em Inglês | MEDLINE | ID: mdl-9223671

RESUMO

We previously reported that the BARF1 (BamH1-A right frame 1) gene product from Epstein-Barr Virus (EBV) may have oncogenic properties since injection into new-born rats of transfected cell lines resulted in the development of BARF1 expressing tumors, which were aggressive in the case of murine fibroblasts and transient in that of human B lymphocytes. As EBV has been associated with nasopharyngeal carcinoma (NPC) and evidence of BARF1 transcription in this cancer was emerging from our biopsy analyses, we examined the effects of BARF1 transfection into primate primary epithelial cells. The expression of the BARF1 open reading frame in primary monkey kidney epithelial cells led us to the establishment of continuously dividing lines. The BARF1 transfectants showed the major characteristics of immortalized cells: morphological change, short cell doubling time, ability to divide at low cell density and continuous growth over 50 passages. Injection of BARF1 transfectants into nude mice did not induce any tumor. Established subclones were shown to be epithelial cells expressing known keratins as well as the BARF1 coded mRNA and protein. This is the first report indicating that expression of the BARF1 gene product in primary epithelial cells may contribute to the establishment of cell lines.


Assuntos
Linhagem Celular , Transformação Celular Viral/genética , Herpesvirus Humano 4/genética , Rim/patologia , Proteínas Virais/genética , Animais , Testes de Carcinogenicidade , Divisão Celular/genética , Células Cultivadas , Epitélio/patologia , Haplorrinos , Queratinas/genética , Queratinas/metabolismo , Camundongos , Camundongos Nus , Fases de Leitura Aberta , Timidina/metabolismo , Transfecção , Proteínas Virais/metabolismo
9.
Clin Cancer Res ; 1(10): 1171-7, 1995 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9815909

RESUMO

Tumor cells become sensitive to the inert prodrug cyclophosphamide (CPA) after transfer of the gene encoding cytochrome P450 2B1. This enzyme activates CPA into 4-hydroxycyclophosphamide, which ultimately degrades into acrolein and phosphoramide mustard, the anticancer and DNA-alkylating metabolite. It is imperative that any prodrug-activating gene therapy strategy against cancer possess the capacity to affect the proliferation of tumor cells even when they do not express the transgene (bystander effect), because current methodologies cannot achieve gene transduction in all tumor cells. Prodrug-activating gene therapy schemes described to date exhibit a bystander effect that is not mediated by conditioned medium in culture and may depend on cell contact. In contrast, we find that CPA-sensitized, P450-expressing C6 glioma cells (C6-P450) transfer cytotoxicity to nonexpressing cells by releasing diffusible metabolites through the medium. A 3-h exposure to the prodrug is necessary and sufficient to achieve killing of the transfected cells, and medium conditioned by these cells can kill untransfected cells with similar potency. This bystander effect occurs in the presence of CPA even when only 10% of cells in culture express the P450 2B1 gene, and it is not reproduced by cells that have been irradiated. In an animal model of intracerebral brain tumors, expression of the P450 2B1 gene within the neoplastic cells enhanced significantly the antitumor effect of CPA, even when it was administered systemically. This study shows that CPA/P450 2B1 gene therapy represents a novel tumor-killing strategy that displays an expanded range of cytotoxic action both spatially and temporally within tumor cells and significantly potentiates the anticancer action of CPA when administered i.v.


Assuntos
Antineoplásicos Alquilantes/farmacologia , Ciclofosfamida/farmacologia , Citocromo P-450 CYP2B1/genética , Terapia Genética/métodos , Pró-Fármacos/farmacologia , Animais , Divisão Celular , Meios de Cultivo Condicionados/farmacologia , Citocromo P-450 CYP2B1/metabolismo , Expressão Gênica , Ratos , Ratos Endogâmicos F344 , Células Tumorais Cultivadas , Ensaio Tumoral de Célula-Tronco
10.
CPT Pharmacometrics Syst Pharmacol ; 4(12): 720-7, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26904386

RESUMO

We present a system of nonlinear ordinary differential equations used to quantify the complex dynamics of the interactions between tumor growth, vasculature generation, and antiangiogenic treatment. The primary dataset consists of longitudinal tumor size measurements (1,371 total observations) in 105 colorectal tumor-bearing mice. Mice received single or combination administration of sunitinib, an antiangiogenic agent, and/or irinotecan, a cytotoxic agent. Depending on the dataset, parameter estimation was performed either using a mixed-effect approach or by nonlinear least squares. Through a log-likelihood ratio test, we conclude that there is a potential synergistic interaction between sunitinib when administered in combination with irinotecan in preclinical settings. Model simulations were then compared to data from a follow-up preclinical experiment. We conclude that the model has predictive value in identifying the therapeutic window in which the timing between the administrations of these two drugs is most effective.

11.
Hum Gene Ther ; 8(17): 2043-55, 1997 Nov 20.
Artigo em Inglês | MEDLINE | ID: mdl-9414253

RESUMO

The identification of transgenes with antitumor activity is critical to the development of gene therapy of cancer. Retrovirus-mediated transfer of the Escherichia coli gpt gene into rat C6 glioma cells without subsequent selection still inhibited the proliferation of this mixed polyclonal population upon addition of the prodrug, 6-thioxanthine, with an ID50 of 4.1 microM, whereas parental C6 cells were not affected at a concentration of 500 microM. In a time-course assay, effects of the prodrug on the mixed polyclonal cell proliferation required at least 10 days of exposure. In mixed co-cultures, a bystander effect was not present over the first 4 days of prodrug exposure, but required trypsinization of the co-cultures and replating at lower densities. This "modified" bystander assay thus revealed a 50% decrease in C6 cell proliferation, even when the initial ratio of gpt-expressing to parental C6 cells was as low as 1:19. In a nude mouse model of subcutaneous tumors, co-grafts of C6 glioma and gpt-retrovirus producer cells displayed retarded growth upon exposure to 6-thioxanthine (6-TX). In a nude mouse model of intracerebral tumors, grafting of the gpt-retrovirus producer cells leads to an 80% reduction in intracerebral tumor volumes after 6-TX treatment. This reduction results in a 28% increase in the mean time of survival of animals that harbor intracerebral tumors (p < 0.0005). These antitumor effects indicate that the gpt/6-TX enzyme/prodrug pair is a promising alternative to the thymidine kinase gene and ganciclovir combination in the gene therapy of cancer.


Assuntos
Antimetabólitos Antineoplásicos/uso terapêutico , Neoplasias Encefálicas/terapia , Escherichia coli/enzimologia , Terapia Genética , Glioma/terapia , Hipoxantina Fosforribosiltransferase/metabolismo , Pró-Fármacos/uso terapêutico , Xantinas/uso terapêutico , Animais , Antimetabólitos Antineoplásicos/toxicidade , Neoplasias Encefálicas/tratamento farmacológico , Modelos Animais de Doenças , Escherichia coli/genética , Estudos de Avaliação como Assunto , Técnicas de Transferência de Genes , Glioma/tratamento farmacológico , Hipoxantina Fosforribosiltransferase/genética , Camundongos , Camundongos Nus , Células Tumorais Cultivadas , Xantinas/toxicidade
12.
Hum Gene Ther ; 6(4): 437-43, 1995 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-7612701

RESUMO

Intratumoral grafting of genetically engineered cells that produce interleukin-4 (IL-4) has been shown to produce tumor regression as well as prolong survival of mice harboring intracerebral gliomas. We sought to determine whether retroviral-mediated gene delivery into tumor cells in situ resulted in enhanced tumor regression by IL-4. Two mouse fibroblast lines were obtained: they both secreted similar levels of IL-4 but one produced a retrovirus vector bearing the IL-4 gene (CRE-MFG-IL-4 cells), whereas the other did not (NIH3T3-IL-4 cells). In mixed transplantation assays in the subcutaneous flanks of athymic mice, CRE-MFG, IL-4 cells were more effective than NIH3T3-IL-4 cells in inhibiting the growth of rat C6 glioma cells (p < 0.005, ANOVA). Subcutaneous tumors injected with fibroblasts that produced a control retrovirus vector without producing IL-4 (CRE-MFG-LacZ cells) did not inhibit subcutaneous tumor growth. An intracranial assay was used to evaluate survival of athymic mice harboring intracranial gliomas. Three days after implanting rat C6 glioma cells into the right frontal lobes of athymic mice, NIH3T3-IL-4 cells (n = 10) or CRE-MFG-IL-4 cells (n = 10) were stereotactically inoculated into the tumor bed. The average survival of mice treated with CRE-MFG-IL-4 cells was 38 days (+/- 2.4, SE), whereas that of mice treated with NIH3T3-IL-4 cells was 31 days (+/- 0.8, SE) (p < 0.005, ANOVA; p < 0.001, log-rank analysis).(ABSTRACT TRUNCATED AT 250 WORDS)


Assuntos
Células 3T3/transplante , Neoplasias Encefálicas/terapia , Terapia Genética , Glioma/terapia , Fatores Imunológicos/uso terapêutico , Interleucina-4/uso terapêutico , Proteínas Recombinantes de Fusão/uso terapêutico , Células 3T3/metabolismo , Células 3T3/virologia , Animais , Neoplasias Encefálicas/patologia , Eosinofilia/etiologia , Lobo Frontal , Vetores Genéticos/genética , Vetores Genéticos/fisiologia , Glioma/patologia , Fatores Imunológicos/administração & dosagem , Fatores Imunológicos/genética , Fatores Imunológicos/metabolismo , Injeções Intralesionais , Interleucina-4/administração & dosagem , Interleucina-4/genética , Interleucina-4/metabolismo , Camundongos , Camundongos Nus , Proteínas Recombinantes de Fusão/administração & dosagem , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Técnicas Estereotáxicas , Replicação Viral
13.
Hum Gene Ther ; 5(2): 183-91, 1994 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8186298

RESUMO

Three vectors derived from retrovirus, herpes simplex virus type 1 (HSV), and adenovirus were compared in cultured rat 9L gliosarcoma cells for gene transfer efficiency and in a 9L rat brain tumor model for histologic pattern and distribution of foreign gene delivery, as well as for associated tumor necrosis and inflammation. At a multiplicity of infection of 1, in vitro transfer of a foreign gene (lacZ from Escherichia coli) into cells was more efficient with either the replication-defective retrovirus vector or the replication-conditional thymidine kinase (TK)-deficient HSV vector than with the replication-defective adenovirus vector. In vivo, stereotactic injections of each vector into rat brain tumors revealed three main histopathologic findings: (i) retrovirus and HSV vector-mediated gene transfer was relatively selective for cells within the tumor, whereas adenovirus vector-mediated gene transfer occurred into several types of endogenous neural cells, as well as into cells within the tumor; (ii) gene transfer to multiple infiltrating tumor deposits without apparent gene transfer to intervening normal brain tissue occurred uniquely in one animal inoculated with the HSV vector, and (iii) extensive necrosis and selective inflammation in the tumor were evident with the HSV vector, whereas there was minimal evidence of tumor necrosis and inflammation with either the retrovirus or adenovirus vectors.


Assuntos
Adenovírus Humanos/genética , Neoplasias Encefálicas/terapia , Vetores Genéticos , Gliossarcoma/terapia , Proteínas Recombinantes de Fusão/biossíntese , Retroviridae/genética , Simplexvirus/genética , Animais , Neoplasias Encefálicas/genética , Neoplasias Encefálicas/patologia , Gliossarcoma/genética , Gliossarcoma/patologia , Inflamação , Masculino , Necrose , Células-Tronco Neoplásicas/metabolismo , Células-Tronco Neoplásicas/microbiologia , Neuroglia/metabolismo , Neuroglia/microbiologia , Neurônios/metabolismo , Neurônios/microbiologia , Ratos , Ratos Endogâmicos F344 , Proteínas Recombinantes de Fusão/genética , Células Tumorais Cultivadas
14.
Hum Gene Ther ; 5(8): 969-78, 1994 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-7948146

RESUMO

Most malignant tumors of the central nervous system do not respond well to chemotherapy. The anticancer drug cyclophosphamide (CPA) is largely ineffective against these neoplasms as its conversion to DNA-alkylating, cytotoxic metabolites is restricted primarily to the liver and these metabolites do not readily cross the blood-brain barrier. Here, we show that brain tumor cells can be sensitized to the cytotoxic effects of CPA, both in culture and in vivo, by introduction of the hepatic enzyme responsible for the activation of CPA, cytochrome P450 2B1. Stable transfection of rat C6 glioma cells with the P450 2B1 gene rendered the cultured tumor cells sensitive to CPA. Further, C6 cells bearing this gene were more sensitive than parental cells to the cytotoxic action of CPA when grown subcutaneously in the flanks of athymic mice. Murine fibroblasts producing a retrovirus vector encoding P450 2B1 and expressing this enzyme were then prepared and grafted into the brains of athymic mice seeded with rat C6 gliomas. Intrathecal administration of CPA prevented the development of meningeal neoplasia and led to partial regression of the parenchymal tumor mass. By contrast, C6 glioma-bearing mice receiving fibroblasts expressing the Escherichia coli lacZ gene and CPA exhibited extensive meningeal tumors and parenchymal solid brain tumors. The in situ activation of CPA by cytochrome P450 2B1 provides a novel approach not only for brain tumor gene therapy, but also for negative, drug-conditional selection of other defined cell populations.


Assuntos
Hidrocarboneto de Aril Hidroxilases , Neoplasias Encefálicas/terapia , Sistema Enzimático do Citocromo P-450/genética , Terapia Genética , Glioma/terapia , Neoplasias Meníngeas/terapia , Esteroide Hidroxilases/genética , Animais , Biotransformação , Neoplasias Encefálicas/tratamento farmacológico , Neoplasias Encefálicas/genética , Células Cultivadas , Terapia Combinada , Ciclofosfamida/farmacocinética , Ciclofosfamida/uso terapêutico , Sistema Enzimático do Citocromo P-450/metabolismo , Resistência a Medicamentos , Escherichia coli , Fibroblastos/transplante , Glioma/tratamento farmacológico , Glioma/genética , Óperon Lac , Neoplasias Meníngeas/tratamento farmacológico , Neoplasias Meníngeas/genética , Camundongos , Camundongos Nus , Ratos , Esteroide Hidroxilases/metabolismo , Transfecção , Células Tumorais Cultivadas
15.
Cancer Gene Ther ; 3(3): 155-62, 1996.
Artigo em Inglês | MEDLINE | ID: mdl-8725879

RESUMO

Genes that encode enzymes that convert inactive "prodrugs" into anticancer metabolites may be therapeutically useful against brain tumors. Unlike other genes tested to date in brain tumor models, the Escherichia coli gpt gene is unique in that it not only sensitizes cells to the prodrug 6-thioxanthine (6TX) but also encodes resistance to a different regimen (mycophenolic acid, xanthine, and hypoxanthine), thus providing a means to select for gpt-positive cells. In the present study, rat C6 glioma cells were infected with a retrovirus vector that transduces this gene. A clonal line (C6GPT-7) was derived that exhibited significant 6TX susceptibility in vitro with an ID50 of 2.5 mumol/L, whereas 50% growth inhibition of parental C6 cells was not achieved at concentrations tested (up to 50 mumol/L). This line also exhibited significant sensitivity to 6-thioguanine (6TG), with an ID50 of 0.05 mumol/L, whereas 50% growth inhibition of parental C6 cells was achieved at 0.5 mumol/L. In a "bystander" assay, C6GPT-7 tumor cells efficiently transferred 6TX sensitivity to C6 cells at ratios as low as 1:9 (C6GPT-7:C6). This in vitro bystander effect was abrogated when C6GPT-7 and C6 cells were separated by a microporous membrane, suggesting that it was not mediated by highly diffusible metabolites. In vivo both 6TX and 6TG significantly inhibited the growth of subcutaneously transplanted C6GPT-7 cells but not that of C6 cells in athymic mice. In an intracerebral model, both 6TX and 6TG exhibited significant antiproliferative effects against tumors formed by C6GPT-7 cells. These findings provide a basis for exploring further gene therapy strategies based on in vivo transfer of the E coli gpt gene to provide chemosensitivity against 6TX and 6TG.


Assuntos
Antimetabólitos Antineoplásicos/toxicidade , Terapia Genética/métodos , Glioma/patologia , Hipoxantina Fosforribosiltransferase/biossíntese , Tioguanina/toxicidade , Transfecção/métodos , Xantinas/toxicidade , Animais , Divisão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Escherichia coli/enzimologia , Escherichia coli/genética , Glioma/tratamento farmacológico , Hipoxantina Fosforribosiltransferase/genética , Hipoxantina Fosforribosiltransferase/metabolismo , Camundongos , Camundongos Nus , Ratos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/metabolismo , Fatores de Tempo , Transplante Heterólogo
16.
World J Gastroenterol ; 3(2): 120, 1997 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-27041968

RESUMO

AIM: To study the method of constructing a pathological model of Piyinxu (spleen-yin deficiency) using Sprague-Dawley rats and to make preliminary observations. METHODS: Folium sennae (0.8 g) and tabellae thyroidei (80 mg) were given daily by stomach feeding to each rat for 12 d to establish the pathological model of Piyinxu. The activity of the rats and the characteristics of their stool were observed. RESULTS: We observed not only the symptoms of Pixu, such as diarrhea, poor appetite, abdominal distention, weight loss, but also Yinxuneire (endogenous heat), such as increase in water drinking, nervousness, and restlessness. These symptoms were all different from that of the control group of Piyangxu (spleen-yang deficiency). CONCLUSION: Combining folium sennae and tabellae thyroidei is an effective method for establishing a pathological model of Piyinxu.

17.
World J Gastroenterol ; 3(3): 188, 1997 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-27239148

RESUMO

AIM: To investigate various characteristics of saliva secreted by patients with TCM-Piyinxu (Spleen-yin deficiency). METHODS: Twenty-five individuals with Piyinxu (15 males and 10 females; age range 26-70 years, mean age = 45 years) diagnosed based on criteria used in traditional Chinese medicine, were compared with 20 individuals with Shenyinxu (Kidney-yin deficiency) (11 males, 9 females; age range 35-75 years, mean age = 50) and 30 normal individuals (17 males, 13 females; age range 35-65 years, mean age = 49 years). After acid stimulation, the saliva flow in each group was measured, and the levels of amylase and protein in saliva were determined using an automatic biochemical analyzer. The resultant data were analyzed using the Kruskal-Wallis test and one-way factorial ANOVA test. RESULTS: The flow rates of saliva and amylase in Piyinxu patients (0.27 ± 0.016 mL/min and 2134.13 ± 343.51 IU/min, respectively) were lower than those in normal subjects (0.46 ± 0.027 mL/min and 3501.63 ± 1099.63 IU/min, respectively, P < 0.01), but higher than those in the Shenyinxu group (0.13 ± 0.051 mL/min and 951.62 ± 383.17 IU/min, respectively, P < 0.01). The three groups showed no significant difference in their level of total salivary protein (Piyinxu group, 3.07 ± 0.60 g/L; Shenyinxu group, 3.01 ± 0.90 g/L, and control group, 2.94 ± 1.13 g/L, P = 0.869), amount of amylase per saliva volume, or their ratio of amylase to protein in secreted saliva (P = 0.173 and P = 0.436, respectively). CONCLUSION: Piyinxu patients showed altered rates of saliva and amylase secretion when compared with those parameters in patients with Shenyinxu and normal subjects.

18.
Parasitol Int ; 49(2): 171-4, 2000 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-10882907

RESUMO

The prevalence of antibodies to Toxoplasma gondii in the sera of rare wildlife in the Shanghai Zoological Garden, PR China, was examined using a modified agglutination test (MAT) and an enzyme-linked immunosorbent assay (ELISA). Forty-one (35%) of 117 animals belonging to two classes, 10 orders, 18 families, 37 genera and 52 species (including sub-species) were sero-positive for MAT. By MAT, T. gondii antibodies were found in 11.1% (4/36) of birds, in 25% (4/16) of primates, in 69.4% (25/36) of carnivores and in 27.6% (8/29) of herbivores. Thirty-three (33.7%) of 98 animals tested by protein A ELISA were sero-positive. By ELISA, T. gondii antibodies were found in none of 36 birds, in 33.3% (4/12) of primates, in 87.1% (27/31) of carnivores and in 10.5% (2/19) of herbivores.


Assuntos
Animais de Zoológico , Anticorpos Antiprotozoários/sangue , Toxoplasma/imunologia , Toxoplasmose/epidemiologia , Toxoplasmose/imunologia , Testes de Aglutinação , Animais , Antígenos de Protozoários/imunologia , China/epidemiologia , Ensaio de Imunoadsorção Enzimática , Prevalência , Estudos Soroepidemiológicos , Toxoplasma/crescimento & desenvolvimento , Toxoplasmose/parasitologia
19.
J Parasitol ; 90(2): 437-8, 2004 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-15165081

RESUMO

Antibodies to Toxoplasma gondii were investigated in serum samples of field mice, Microtus fortis, from Yuanjiang, Hunan Province, People's Republic of China. The modified agglutination test (MAT) incorporating formalin-fixed whole tachyzoites and mercaptoethanol was used to determine antibodies. Antibodies to T. gondii (MAT > or = 1:20) were found in 36 (29%) of 124 trapped mice. The antibody titers of positive sera (percentage in parentheses) were 1:20 (8.9), 1:40 (3.2), 1:80 (3.2), 1:160 (1.6), 1:320 (1.6), 1:640 (1.6), 1:1,280 (1.6), 1: 2,560 (0.8), and > 1:2,560 (6.5). No antibody to T. gondii was found in 104 sera of laboratory-bred M. fortis infected with Schistosoma japonicum between 1 and 45 days after infection.


Assuntos
Anticorpos Antiprotozoários/sangue , Arvicolinae/parasitologia , Doenças dos Roedores/epidemiologia , Toxoplasma/imunologia , Toxoplasmose Animal/epidemiologia , Testes de Aglutinação/veterinária , Animais , China/epidemiologia , Doenças dos Roedores/parasitologia , Sensibilidade e Especificidade , Estudos Soroepidemiológicos
20.
Artigo em Zh | MEDLINE | ID: mdl-12567476

RESUMO

OBJECTIVE: To evaluate the diagnostic value of three agglutination tests used in three countries for detection of antibodies to Taxoplasma gondii. METHODS: A total of 288 human serum samples were assayed using modified agglutination test (MAT-1, using selfmade antigen), latex agglutination test (LAT, using Japanese Kit) and modified agglutination test (MAT-2, using French antigen). RESULTS: The positive rates of MAT-1 (> or = 1:20), LAT (> or = 1:32) and MAT-2 (> or = 1:20) were 9.7% (28/288), 8.9% (10/112) and 8.1% (17/210), respectively. No significant statistical difference was found among these positive rates (chi 2 = 0.392, P > 0.05). High agreements were found between MAT-1 and LAT (93.7%), LAT and MAT-2 (94.5%), and MAT-2 and MAT-1 (97.3%). Significant correlation were demonstrated in MAT-1 and LAT (r = 0.613), LAT and MAT-2 (r = 0.551), and MAT-2 and MAT-1 (r = 0.841), p < 0.001. CONCLUSION: The detection efficiency of the three agglutination tests is in good agreement and could alternatively be used for the diagnosis of toxoplasmosis.


Assuntos
Testes de Aglutinação/métodos , Anticorpos Antiprotozoários/sangue , Toxoplasma/imunologia , Toxoplasmose/diagnóstico , Animais , Humanos , Sensibilidade e Especificidade
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