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1.
J Cardiovasc Electrophysiol ; 31(4): 960-963, 2020 04.
Artigo em Inglês | MEDLINE | ID: mdl-32077548

RESUMO

We present a case of wide-complex tachycardia in which the clinical electrophysiological diagnosis was considered to be bundle branch re-entry ventricular tachycardia. A series of ventricular entrainment attempts were performed from the left and right ventricular septum to confirm the diagnosis. Entrainment pacing with a general current output (10 mA) was performed from the right ventricular septum with manifest fusion and a post-pacing interval similar to tachycardia cycle length. Thereafter, another entrainment attempt with a greater current output (20 mA) was performed from the same site. Paradoxically, concealed fusion was demonstrated by selective RB capture only, though there was no clear "RB" potential seen. In this case, we attempt to explain and illustrate the mechanism of paradoxical near-field inability to capture with increasing current strength.


Assuntos
Potenciais de Ação , Fascículo Atrioventricular/fisiopatologia , Estimulação Cardíaca Artificial , Eletrocardiografia , Técnicas Eletrofisiológicas Cardíacas , Frequência Cardíaca , Taquicardia Ventricular/diagnóstico , Adulto , Feminino , Humanos , Valor Preditivo dos Testes , Taquicardia Ventricular/fisiopatologia , Fatores de Tempo
2.
Heart Rhythm ; 13(7): 1460-7, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-26961304

RESUMO

BACKGROUND: In patients with idiopathic left ventricular tachycardia (ILVT), the arrhythmogenic substrate is poorly understood. OBJECTIVE: The purpose of this study was to elucidate the ILVT characteristics and outcome of radiofrequency catheter ablation in patients with ILVT. METHODS: Twenty-four patients with ILVT and 15 patients with left accessory pathways (control) underwent high-density mapping of the left His-Purkinje system during sinus rhythm (SR) using 3-dimensional electroanatomic mapping. RESULTS: Fragmented antegrade Purkinje potential (FAP) was represented at the left ventricular septum slightly inferoposterior to the left posterior fascicle (LPF) in 23 patients with ILVT. In control subjects, no FAPs could be recorded at the same region, FAPs were identified at the proximal portion of the LPF (4 patients) and at the distal LPF (1 patient). The finding of any FAPs in ILVT patients was significantly higher than that in control patients (23/24 vs 5/15, P < .01). Radiofrequency ablation at the area of FAP resulted in successful ablation in 23 patients with ILVT. No ILVT recurred during follow-up of 16.3 ± 7.2 months. CONCLUSION: In patients with ILVT, FAP located at the left ventricular septum slightly inferoposterior to the LPF is a novel finding using 3-dimensional electroanatomic mapping. The FAP may represent an arrhythmogenic substrate in ILVT and may be used for guiding successful ablation.


Assuntos
Mapeamento Potencial de Superfície Corporal/métodos , Ventrículos do Coração , Taquicardia Ventricular , Adolescente , Adulto , Ablação por Cateter/métodos , Ecocardiografia Tridimensional/métodos , Técnicas Eletrofisiológicas Cardíacas/métodos , Feminino , Ventrículos do Coração/patologia , Ventrículos do Coração/fisiopatologia , Humanos , Masculino , Avaliação de Processos e Resultados em Cuidados de Saúde , Ramos Subendocárdicos , Reprodutibilidade dos Testes , Cirurgia Assistida por Computador/métodos , Taquicardia Ventricular/diagnóstico , Taquicardia Ventricular/etiologia , Taquicardia Ventricular/fisiopatologia
3.
BMC Cardiovasc Disord ; 15: 98, 2015 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-26324443

RESUMO

BACKGROUND: To explore the association between atrial fibrillation (AF) and serum uric acid (SUA) in a general population in rural China. METHODS: From January 2013 to August 2013, we performed a cross-sectional study involving 11,956 permanent residents ≥ 35 years old in the rural Liaoning province of China. All participants completed a questionnaire, had a physical examination, and underwent an electrocardiogram (ECG) and echocardiogram. AF was diagnosed from ECG findings and/or a history of physician-confirmed AF. Blood samples were drawn for laboratory analyses and hyperuricemia was defined as an SUA level > 7.0 mg/dL in men and > 5.7 mg/dL in women, based on the NHANES-III laboratory definition. Logistic regression analyses were performed to estimate the crude and independent associations between hyperuricemia and the prevalence of AF. RESULTS: A total of 139 participants were diagnosed with AF, of which, 72 were self-reported, 45 were ECG-diagnosed, and 22 were both. There was a higher prevalence of AF in participants with hyperuricemia than those with normal SUA levels (2.4 vs. 1.0 %; P < 0.001). The odds ratios (OR) and 95 % confidence intervals (CI) were 2.37 (1.61-3.49) when compared to participants with normal SUA. After adjustment for other cardiovascular and AF risk factors, the independent association remained (OR = 1.94, 95 % CI: 1.26-3.00). Similar associations were observed between SUA as a continuous variable and AF prevalence (adjusted OR = 1.20, 95 % CI: 1.06-1.36). The independent associations were significant in men (Ps < 0.05) but not in women (Ps > 0.05), although the interaction logistic regression analyses presented these differences as not being statistically significant (Ps > 0.05). CONCLUSIONS: SUA is positively associated with the prevalence of AF in rural China.


Assuntos
Fibrilação Atrial/complicações , Fibrilação Atrial/epidemiologia , Hiperuricemia/complicações , Adulto , Fatores Etários , China/epidemiologia , Estudos Transversais , Eletrocardiografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Fatores de Risco , População Rural , Fatores Sexuais
4.
Int J Cardiol ; 182: 13-7, 2015 Mar 01.
Artigo em Inglês | MEDLINE | ID: mdl-25576710

RESUMO

OBJECTIVES: To evaluate the prevalence of atrial fibrillation (AF) in physical laborers in rural China and identify contributing risk factors. METHODS: A cross-sectional study of 11,956 permanent residents of Liaoning Province in rural China≥35y of age (primarily physical laborers) was conducted between January and August 2013 (response rate 85.3%). All participants completed a questionnaire and underwent a physical exam, echocardiography and electrocardiography. Blood samples were drawn for laboratory analyses, and AF was diagnosed on the basis of history and electrocardiograph findings. Risk factors for AF were evaluated with a stepwise logistic regression analysis. RESULTS: The prevalence of AF was 1.2% overall, but rose steeply with age (0.1% in those 35-44y of age, and 4.6% in those≥75y); there was no significant gender difference at any age. Independent risk factors for AF were age (odds ratio [OR] 1.89; P<0.001), diabetes (OR 2.07; P=0.001), history of myocardial infarction (OR 5.91; P<0.001), low left ventricular ejection fraction (OR 1.85; P=0.005), and low physical activity (OR 1.72; P=0.003), whereas obesity, hypertension, cholesterol and triglyceride levels, current smoking and drinking, left ventricular hypertrophy, and family history of AF were not significant contributors. CONCLUSIONS: Although the prevalence of AF in physical labors in rural China is low, age, diabetes, history of myocardial infarction, low left ventricular ejection fraction, and low physical activity are independent risk factors.


Assuntos
Fibrilação Atrial/epidemiologia , Medição de Risco , População Rural , Adulto , Fibrilação Atrial/diagnóstico , China/epidemiologia , Estudos Transversais , Eletrocardiografia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Razão de Chances , Prevalência , Estudos Retrospectivos , Fatores de Risco
5.
J Mol Evol ; 67(5): 448-64, 2008 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-18850061

RESUMO

The resolution potential of internal transcribed spacer 2 (ITS2) at deeper levels remains controversial. In this study, 105 ITS2 sequences of 55 species in Calyptratae were analyzed to examine the phylogenetic utility of the spacer above the subfamily level and to further understand its evolutionary characteristics. We predicted the secondary structure of each sequence using the minimum-energy algorithm and constructed two data matrixes for phylogenetic analysis. The ITS2 regions of Calyptratae display strong A-T bias and slight variation in length. The tandem and dispersed repeats embedded in the spacers possibly resulted from replication slippage or transposition. Most foldings conformed to the four-domain model. Sequence comparison in combination with the secondary structures revealed six conserved motifs. Covariation analysis from the conserved motifs indicated that the secondary structure restrains the sequence evolution of the spacer. The deep-level phylogeny derived from the ITS2 data largely agreed with the phylogenetic hypotheses from morphologic and other molecular evidence. Our analyses suggest that the accordant resolutions generated from different analyses can be used to infer deep-level phylogenetic relations.


Assuntos
DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Dípteros/genética , Evolução Molecular , Filogenia , Algoritmos , Animais , Sequência de Bases , Teorema de Bayes , Sequência Conservada , Dípteros/classificação , Especiação Genética , Sequências Repetitivas Dispersas/genética , Modelos Moleculares , Dados de Sequência Molecular , Polimorfismo Genético , Alinhamento de Sequência , Sequências de Repetição em Tandem/genética
6.
Mol Phylogenet Evol ; 37(2): 313-26, 2005 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-16137895

RESUMO

Pentatomomorpha is the second suborder in size only to Cimicomorpha in Heteroptera. However, the phylogenetic relationships among members of the suborder are not well established. Sequences from partial nuclear ribosomal 18S gene and mitochondrial COX1 gene were analyzed separately and in combination to generate a preliminary molecular phylogeny of Pentatomomorpha based on 40 species representing 17 putative families. Analyses of the combined sequence data provided a better-resolved and more robust hypothesis of Pentatomomorpha phylogeny than did separate analyses of the individual genes. The phylogenies were mostly congruent with morphological studies. Results strongly supported the monophyly of the infraorder Pentatomomorpha, and the placement of Aradoidea as sister to Trichophora. The monophyletic Trichophora was grouped into two major lineages, one being the superfamily Pentatomoidea, and the other comprising Lygaeoidea, Coreoidea, and Pyrrhocoroidea. The analysis of the ML and ME trees of combined dataset supported the monophyletic Pentatomoidea. In all analysis the Pyrrhocoroidea was polyphyletic; the monophyletic Lygaeoidea was supported only in the analysis of ME tree, and Coreoidea was polyphyletic except in the MP tree of combined dataset. The molecular and morphylogical data both indicated that the family Coreoidae should be revised subsequently. Our phylogenetic results suggested that the COX1 segment alone might not be an optimal molecular marker for the phylogeny of Pentatomomorpha.


Assuntos
Ciclo-Oxigenase 1/genética , DNA Mitocondrial/genética , DNA Ribossômico/genética , Heterópteros/classificação , Filogenia , RNA Ribossômico 18S/genética , Animais , Núcleo Celular/genética , DNA Mitocondrial/análise , DNA Ribossômico/análise , Heterópteros/genética , RNA Ribossômico 18S/análise
7.
Sheng Wu Gong Cheng Xue Bao ; 20(5): 800-4, 2004 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-15974013

RESUMO

In order to produce recombinant human anti-HBsAg Fab antibody in Pichia pastoris, the recombinant yeast was fermented using fed-batch system in a 30 L bioreactor. The fermentation temperature was 30 degrees C, the pH was 5.0 approximately 5.3, and the DO was 20% approximately 30%. The recombinant Fab antibody was purified from crude culture supernatant by ion exchange and analyzed by SDS-PAGE and western blot and ELISA. When the absorbance (OD600) of broth reach 300 at the end of fed-batch phase, the induced phase was initiated. The results showed that recombinant human anti-HBsAg Fab antibody was high-level expressed in recombinant Pichia pastoris using a fed-batch fermentation system. Both chains of the Fab were successfully expressed upon methanol induction. After 192 h of induction, the expression level of recombinant Fab (soluble) reached 412 mg/L. The recombinant Fab antibody was purified effectively by ion-exchange chromatography from the fermentation supernatant to a purity of 95%. And the affinity activities of the purified recombinant Fab antibdy and fermentation supernatant were detected, and both of them showed high affinity activities. The results demonstrated that recombinant human anti-HBsAg Fab antibody could be high level produced by fed-batch fermentations in Pichia pastoris. Which can be efficiently used in industrial production.


Assuntos
Fermentação , Anticorpos Anti-Hepatite B/biossíntese , Antígenos de Superfície da Hepatite B/imunologia , Fragmentos Fab das Imunoglobulinas/biossíntese , Pichia/genética , Proteínas Recombinantes/biossíntese , Anticorpos Anti-Hepatite B/isolamento & purificação , Humanos , Fragmentos Fab das Imunoglobulinas/isolamento & purificação , Proteínas Recombinantes/isolamento & purificação
8.
J Virol Methods ; 114(2): 145-50, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14625049

RESUMO

Pseudorabies virus glycoprotein E (PRV gE) has been recognized as a suitable diagnostic antigen for pseudorabies. In order to produce gE antigen in large quantities and at low cost, a gene fragment encoding PRV gE epitopes was expressed in Pichia pastoris expression system. SDS-PAGE and Western blotting revealed that the expression product was two recombinant proteins, approximately 38 and 32 kDa, in the culture supernatant of P. pastoris integrant 72 h after induction. Protein concentration assay showed the expression product amounted to 106.7 mg/l, accounting for 66.67% of total culture supernatant proteins. An indirect PRV gE-ELISA was then established by using the recombinant expression product as a coating antigen. Cross-reactivity assay showed that this antigen was PRV specific. Reproducibility experiment displayed good consistency. Comparison of detection results of 348 field serum samples between PRV gE-ELISA and a commercially available PRV diagnostic kit showed there was no significant difference between these two methods (P > 0.05).


Assuntos
Anticorpos Antivirais/sangue , Herpesvirus Suídeo 1/imunologia , Pichia/metabolismo , Pseudorraiva/diagnóstico , Proteínas Recombinantes/imunologia , Proteínas do Envelope Viral/metabolismo , Animais , Antígenos Virais/genética , Antígenos Virais/imunologia , Antígenos Virais/metabolismo , Western Blotting , Eletroforese em Gel de Poliacrilamida , Ensaio de Imunoadsorção Enzimática , Epitopos , Herpesvirus Suídeo 1/genética , Herpesvirus Suídeo 1/metabolismo , Pichia/genética , Pseudorraiva/virologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Reprodutibilidade dos Testes , Suínos , Proteínas do Envelope Viral/genética , Proteínas do Envelope Viral/imunologia
9.
Zhonghua Zheng Xing Wai Ke Za Zhi ; 19(4): 285-7, 2003 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-14628420

RESUMO

OBJECTIVE: Despite the causes for melanin increase, the increased gene expression of TYR is a common pathological process. Based on this viewpoint, antisense-S-Oligo of TYR was designed and synthesized to regulate synthesis of melanin in order to explore the treatment for skin pigmentation. METHODS: The cultured melanocytes were divided into 3 groups. The group 1 was treated with endothelin, group 2 treated with ultraviolet ray and group 3 was used as the control. In each group, the 5' antisense-S-Oligo, the 3' antisense-S-Oligo, the mixed antisense-S-Oligo of TYR or Dotap only was added. The melanin content and TYR gene expressions were examined. RESULTS: The 5' antisense-S-Oligo, the 3' antisense-S-Oligo and the mixed antisense-S-Oligo significantly inhibited the increase of melanin content and TYR gene expression, which were caused by endothelin or ultraviolet ray treatment. Of the three treatments, the 3' antisense-S-Oligo showed the strongest effect. CONCLUSION: Antisense-S-Oligo has significant regulating effects on TYR gene expression and melanin content. The 3' antisense-S-Oligo is more effective than the 5' antisense-S-Oligo.


Assuntos
Expressão Gênica , Melaninas/biossíntese , Oligodesoxirribonucleotídeos Antissenso/farmacologia , Tirosina/genética , Região 3'-Flanqueadora/genética , Região 5'-Flanqueadora/genética , Endotelinas/farmacologia , Melanócitos/efeitos dos fármacos , Melanócitos/metabolismo , Melanócitos/efeitos da radiação , Oligodesoxirribonucleotídeos Antissenso/genética , Tirosina/metabolismo , Raios Ultravioleta
10.
Artigo em Chinês | MEDLINE | ID: mdl-12958657

RESUMO

The p74 gene of Autographa californica multicasid nucleopolyhedrovirus (AcMNPV) bacmid was knockouted and substituted by the p74 gene of Spodoptera litura multicapsid nucleopolyhedrovirus (SpltMNPV), using RecA-mediated homologous recombination in the E. coli. No selection marker, which might influence the expression and function of p74 gene, was left in the modified p74 locus. The promoter of AcMNPV p74 gene directly controlled the expression of SpltMNPV p74 gene in the recombinant AcMNPV bacmid-polhSL74. RT-PCR showed that the substituted p74 gene was transcribed. Bioassay showed that the recombinant virus AcMNPV bacmid-polhSL74 could not infect the Argyrogramma agnata larvae per os, and thus showing the p74 gene is species-specific.


Assuntos
Nucleopoliedrovírus/genética , Proteínas do Envelope Viral/genética , Animais , Sequência de Bases , Regulação Viral da Expressão Gênica , Larva/virologia , Mariposas/virologia , Mutação , Nucleopoliedrovírus/crescimento & desenvolvimento , Plasmídeos/genética , Recombinação Genética , Especificidade da Espécie
11.
Parasitol Res ; 90(4): 287-93, 2003 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12884021

RESUMO

We present an analysis of the expression profile of Schistosoma japonicum cercariae by a strategy based on expressed sequence tags (ESTs). A cDNA library from S. japoniucm cercariae was constructed and was used to generate ESTs. In total, 201 clones randomly selected from the library were sequenced; 136 ESTs were successfully obtained and sent to the BLAST server for homology searching. Among the 136 ESTs, 85 (62.50%) did not match any protein or gene sequence published in the BLAST databases; these comprised 75 (55.15%) ESTs matched (or partly matched) S. japonicum and Schistosoma mansoni ESTs, 4 (2.94%) matched human ESTs and 6 (4.41%) that did not match any sequence. Fifty-one (37.50%) ESTs were identified by the BLAST server; these consisted of 8 (5.88%), 9 (6.62%) and 34 (25.00%) that showed high homology with genes or proteins reported from S. japonicum, S. mansoni and other organisms, respectively. These identified ESTs can be grouped into nine categories: transporters (1.96%), secretory proteins (1.96%), kinases (3.92%), proteases (5.88%), structural and cytoskeletal proteins (13.73%), metabolism-related proteins (9.80%), regulatory and signaling proteins (11.76%), transcription and translation machinery (25.50%), and others (25.50%). Several interesting new genes cloned from this cDNA library are discussed here. These findings will be valuable for the understanding of the biology of this parasite.


Assuntos
Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Genes de Helmintos , Schistosoma japonicum/genética , Animais , DNA Complementar/genética , Bases de Dados de Ácidos Nucleicos , Regulação da Expressão Gênica , Biblioteca Gênica , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Schistosoma japonicum/crescimento & desenvolvimento , Schistosoma japonicum/metabolismo , Análise de Sequência de DNA
12.
Int J Cancer ; 106(1): 60-5, 2003 Aug 10.
Artigo em Inglês | MEDLINE | ID: mdl-12794757

RESUMO

Nonrandom allelic loss on chromosome 3p is a common event in nasopharyngeal carcinoma (NPC) with the implication that certain tumor suppressor gene(s) in this region are involved in the pathogenesis of these tumors. The BLU gene, located at 3p21.3, has recently been identified as a candidate tumor suppressor gene due to the occurrence of missense mutations and loss of its expression in lung cancer. To investigate the involvement of BLU gene in NPC, we examined both genetic and epigenetic changes of BLU in NPC primary tumors and cell lines. No pathogenic mutations were detected in the entire coding region of this gene in 45 primary NPC tumors and 5 NPC cell lines. While BLU was expressed in 100% (15 of 15) of noncancerous nasopharyngeal epithelia, its transcripts were missing in all 5 NPC cell lines, and absent or reduced mRNA levels were observed in 78% (28 of 36) of the primary tumors. In the NPC cell lines, loss of BLU expression correlated with hypermethylation of the CpG island promoter sequence, and expression was restored after treatment with 5'-aza-2'-deoxycytidine. Methylation specific PCR analysis revealed that the BLU promoter was highly methylated in 74% (17 of 23) of primary tumors in which BLU was downregulated, whereas only 2 of 9 non-neoplastic nasopharyngeal epithelia exhibited hypermethylation in the BLU promoter region. The high incidence of BLU alterations suggests that it may be one of the critical tumor suppressor genes on chromosome 3p21.3 involved in the development of NPC.


Assuntos
Azacitidina/análogos & derivados , Carcinoma/genética , Cromossomos Humanos Par 3 , Neoplasias Nasofaríngeas/genética , Proteínas/genética , Adulto , Idoso , Azacitidina/farmacologia , Sequência de Bases , Ilhas de CpG , Proteínas do Citoesqueleto , Metilação de DNA , Primers do DNA/farmacologia , Decitabina , Inibidores Enzimáticos/farmacologia , Genes Supressores de Tumor , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Mutagênicos , Mutação , Reação em Cadeia da Polimerase , Regiões Promotoras Genéticas , Biossíntese de Proteínas , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Sulfitos/farmacologia , Células Tumorais Cultivadas , Proteínas Supressoras de Tumor
13.
Artigo em Chinês | MEDLINE | ID: mdl-12673392

RESUMO

The gene of LIP1, the most important isoenzyme of Candida rugosa lipase (CRL), was artificially synthesized according to its mature peptide sequence. It consisted of 20 codons of preference in Pichia pastoris. The artificial gene was cloned into methanol-inducible expression vector pPICZalphaA, and constitutive expression vector pGAPZalphaA, respectively. The linearized recombinant plasmids were transformed into chromosome of Pichia pastoris SMD1168H strain by electroporation. The abilities of expressing LIP1 in both transformed yeasts had been compared, and the yeast transformed with pGAPZalphaA was more efficient than pPICZalphaA. A recombinant yeast strain named CHT-II expressed LIP1 constitutively, and was the most efficient one. Some enzymatic properties of the recombinant LIP1 were also determined. CHT-II secreted LIP1 into supernate at a level of 2.00x10(5) u/L after 72 h (the cells had been transferred to fresh culture medium at the 24th h). After optimizing the conditions for high cell-density fermentation, the selected yeast strain could secrete LIP1 into supernate at a level of 1.395x10(6) u/L after 72 h. The results indicated that this modification of lip1 gene was successful.


Assuntos
Candida/enzimologia , Lipase/genética , Pichia/genética , Sequência de Aminoácidos , Eletroforese em Gel de Poliacrilamida , Fermentação , Regulação Enzimológica da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Genes Sintéticos/genética , Concentração de Íons de Hidrogênio , Lipase/metabolismo , Dados de Sequência Molecular , Pichia/metabolismo , Plasmídeos/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Especificidade por Substrato , Temperatura , Transformação Genética , Trioleína/metabolismo
14.
Ai Zheng ; 22(1): 16-20, 2003 Jan.
Artigo em Chinês | MEDLINE | ID: mdl-12561429

RESUMO

BACKGROUND & OBJECTIVE: Though the molecular etiology of nasopharyngeal carcinoma(NPC) is currently unknown, evidence from both loss of heterozygosity analysis and functional studies suggested that there are NPC-associated tumor suppressor genes(TSGs) residing in chromosome 3p21.3. Recently, two members of semaphorin family, SEMA3B and SEMA3F gene, located at 3p21.3, were characterized as TSGs. Studies showed that SEMA3B and SEMA3F are capable of suppressing the growth of tumor cells and inducing apoptosis. Loss of SEMA3B mRNA expression or aberrant SEMA3F cellular localization were found in lung cancers. In order to investigate the involvement of SEMA3B and SEMA3F in NPC, the authors examined both mutation and expression of these two genes in NPC. METHODS: The entire coding regions, the splice donor/acceptor sites, and partial regulatory regions of SEMA3B and SEMA3F gene were screened for mutations by PCR-sequencing in 21 primary NPC tumors and 2 NPC cell lines(CNE2 and SUNE1). The mRNA expression levels were determined by semi-quantitative RT-PCR analysis. RESULTS: No somatic mutation was found in either SEMA3B or SEMA3F gene. However, two missense polymorphisms including Thr415Ile and lle242Met were found in SEMA3B in NPC. For the Thr415Ile polymorphism, the Ile allele type which leads to SEMA3B function defects was predominant in NPC with the allele frequency of 64% (27/42). SEMA3B mRNA was expressed in all 6 non-neoplastic nasopharyngeal epithelia, but was absent or down-regulated in 76% (16/21) of primary NPC tumors. No significant difference of SEMA3B expression was observed between NPC and noncancerous controls. CONCLUSION: High frequency of SEMA3B expression alterations suggests that the inactivation of this gene was strongly associated with NPC. SEMA3B may be a tumor suppressor on 3p21.3 involved in NPC.


Assuntos
Genes Supressores de Tumor , Glicoproteínas de Membrana/biossíntese , Proteínas de Membrana/biossíntese , Mutação de Sentido Incorreto , Neoplasias Nasofaríngeas/metabolismo , Proteínas do Tecido Nervoso/biossíntese , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Linhagem Celular Tumoral , Cromossomos Humanos Par 3/genética , Éxons , Humanos , Glicoproteínas de Membrana/genética , Proteínas de Membrana/genética , Neoplasias Nasofaríngeas/patologia , Proteínas do Tecido Nervoso/genética , Semaforinas
15.
Ai Zheng ; 22(2): 128-35, 2003 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-12600284

RESUMO

BACKGROUND & OBJECTIVE: Nonrandom allelic loss at chromosome 3p21.3 is a common and early event in nasopharyngeal carcinoma (NPC), which implicates the presence of tumor suppressor genes (TSGs) that may be involved in the pathogenesis of NPCs. BLU gene, containing a MYND domain and located at 3p21.3, has been considered as a NPC associated candidate tumor suppressor gene (TSG) due to the occurrence of loss of its expression and aberrant promoter hypermethylation in most NPCs. This study was designed to construct expression vectors containing either wild type BLU gene and its mutants and to analyze the effect of BLU gene on proliferation of NPC cells by transfection assays. METHODS: The full-length cDNA of BLU gene was amplified by RT-PCR. The expression vectors containing various BLU mutants were constructed by site-directed mutagenesis by overlapping PCR. These mutants include a MYND domain deletion mutant, a Ser402Phe and del405Cys, del406Ser mutant, and a Gly160Arg mutant. The wild type BLU gene and the MYND domain deletion mutant were transfected into NPC cell lines CNE1 and CNE2. The effect on apoptosis was determined by TUNEL assay. Cellular proliferation of the stably-transfected cells was examined with cell growth curve and by colony formation assays. Tumorigenicity in nude mice of CNE2 stably-transfected with BLU was investigated. RESULTS: No significant difference in apoptosis index (AI) was observed between cells transfected with wild type or MYND domain deleted BLU gene and cells transfected with plasmid controls. Exogenous expression of wild type BLU gene had no effect on growth rate and colony formation ability of CNE1 and CNE2. BLU gene showed no suppressor ability in CNE2 tumorigenicity. CONCLUSION: Although BLU gene was frequently altered in NPCs, its suppressor role in NPC cells proliferation was not evident. Thus, the possibility of BLU gene as a TSG involved in NPC development remained to be elucidated by further studies.


Assuntos
Apoptose , Proteínas/farmacologia , Animais , Divisão Celular/efeitos dos fármacos , Mapeamento Cromossômico , Cromossomos Humanos Par 3 , Proteínas do Citoesqueleto , Modelos Animais de Doenças , Genes Supressores de Tumor , Humanos , Marcação In Situ das Extremidades Cortadas , Camundongos , Camundongos Nus , Mutação , Neoplasias Nasofaríngeas/patologia , Transplante de Neoplasias , Neoplasias Experimentais/tratamento farmacológico , Proteínas/genética , Proteínas/uso terapêutico , Transfecção , Células Tumorais Cultivadas , Proteínas Supressoras de Tumor
16.
Sheng Wu Gong Cheng Xue Bao ; 18(5): 546-50, 2002 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-12561196

RESUMO

Using of two-step integrating technology, transducted the H and L chain gene of humanized Fab fragment of anti-HB-sAg antibody into the genome of methylotropic yeast P. pastoris. Constructed a engineering yeast to produce humanized Fab fragment of the anti-HBsAg antibody. The Fab fragment was efficiently secreted into the medium at a concentration of 50-80 mg/L. The Fab fragment was purified from culturing supernatant of the recombinant yeas by affinity chromatography. The ELISA analysis showed the high affinity of the expressed humanized Fab fragment to the HBsAg.


Assuntos
Anticorpos Anti-Hepatite B/genética , Antígenos de Superfície da Hepatite B/imunologia , Fragmentos Fab das Imunoglobulinas/genética , Pichia/genética , Proteínas Recombinantes/biossíntese , Cromatografia de Afinidade , Clonagem Molecular , Ensaio de Imunoadsorção Enzimática , Anticorpos Anti-Hepatite B/biossíntese , Anticorpos Anti-Hepatite B/isolamento & purificação , Humanos , Fragmentos Fab das Imunoglobulinas/biossíntese , Fragmentos Fab das Imunoglobulinas/isolamento & purificação , Proteínas Recombinantes/isolamento & purificação
17.
Di Yi Jun Yi Da Xue Xue Bao ; 21(11): 809-811, 2001.
Artigo em Inglês | MEDLINE | ID: mdl-12426177

RESUMO

OBJECTIVE: To understand the general profiles of the genes included in Schistosoma japonicum (Sj) cercarriae cDNA library that we have constructed for the purpose of identifying novel genes. METHODS: The phages in Sj cercariae cDNA library were transformed into plasmids after they had invaded into E.coli BM25.8, and the E.coli clones containing the plasmids were cultured in LB medium supplemented with aminobenzylpenicillin. The isolated colonies were selected for plasmid extraction and the DNAs subsequently extracted from the plasmids underwent sequence analysis with a sequencing primer to obtain the expressed sequence tags (ESTs). After sequence editing of the resulted ESTs, comparison of these sequences with those listed in the GenBank database was performed for homology analysis. The new sequences generated from the analysis were submitted to GenBank and the accession numbers acquired. RESULTS and CONCLUSION :Altogether 58 ESTs were identified among which 3 possed highly homologous nucleotide sequences with the known genes of Sj, 2 showed the homology with Schistosoma mansori genes, 7 had the homologous protein sequences, but not the nucleotide sequences, with other species, and another 27 were poorly homologous in terms of both the nucleotide and protein sequences.

18.
Artigo em Inglês | MEDLINE | ID: mdl-12058189

RESUMO

A 4 730 bp region of the Spodoptera litura Nucleopolyhedrovirus genome EcoRI-E fragment was sequenced, in which the odv-e66 gene and other two ORFs, ORF 1086 and the p34 gene, were found. The SpltNPV odv-e66 gene open reading frame was deduced to be 2 079 bp, encoding a protein of 692 amino acids. Unlike odv-e66 genes of AcMNPV and LsNPV, only one transcriptional initiation conserved sequence ATAAG of the late gene was present upstream of the initial codon ATG of SpltNPV odv-e66 gene, and no poly(A) signal was detected at the 3' end of the gene. Four highly conserved regions homologous to those of LdNPV, LsNPV, AcMNPV, BmNPV, CfNPV and OpNPV were present in the amino acid sequence of SpltNPV ODV-E66 protein, with the putative nuclear targeting signal(NTS) KKIWAEDGR in the fourth conserved region. The ORF upstream of the odv-e66 gene had the same direction of odv-e66 gene, but different reading frame. Besides the CAAT box, the early gene regulatory element ACGT and GC motifs and the transcriptional initiation conserved sequence TTAAG of the late gene were present at the 5' end of this ORF. A putative protein of 289 amino acid residues with Mr. about 34 kD was encoded by the ORF, so that it is named P34 protein. A leucine zipper and a leucine zipper-like structure were found in the C- and N-termini of SpltNPV P34 protein respectively. The functions of those regulatory elements may be of great importance to SpltNPV genome. An 1 086 nt open reading frame (ORF) capable of encoding a 361 amino acids polypeptide was found at the 3' end of the odv-e66 gene and in opposite direction. The ORF 1086 protein shares 44% amino acid sequence homology with AcMNPV ORF 109.

19.
Artigo em Inglês | MEDLINE | ID: mdl-12167986

RESUMO

The p10 gene was localized within the 3.0 kbp BamH I fragment of the SINPV genome. With a coding sequence of 318 nucleotides, corresponding to a protein of 105 amino acids, the p10 gene of SINPV is the longest p10 gene identified so far in NPVs. The SINPV p10 gene is also distinct in having two A/TTTGTA motifs in the promoter region. With 10 heptad repeats, the SINPV P10 protein is probably forming a relatively large coiled-coil structure. The organization of SINPV ORF552-p10 gene-ORF945 cluster is collinear with that of SpliNPV and their corresponding amino acid sequences of each ORF share high similarity. This indicates that SINPV and SpliNPV belong to a homologous baculovirus group.

20.
Artigo em Inglês | MEDLINE | ID: mdl-12174264

RESUMO

A refractile body antigen Etp28 Gene of Eimeria tenella (Guangdong Strain) sporozoites was cloned by PCR from the synthesized first strand cDNA. It has a high homology with the previously reported Etp28 gene of Merck Strain LS18. The gene was expressed through standard procedures in the modified Autographa californica nuclear polyhedrosis virus (AcMNPV-OCC(-)) expression system and large amount of heterologous fusion protein (GST-6xHis-Etp28) was obtained. The expression product was about 21.3% of the total cellular protein of an infected cell and corresponding to approximately 0.42 mg of recombinant protein per 10(6) cells. And the immunoprotective trial showed that this candidate for recombinant vaccine conferred partial protection against chicken coccidiosis.

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