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1.
Medicine (Baltimore) ; 103(16): e37848, 2024 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-38640332

RESUMO

OBJECTIVE: To investigate the clinical efficacy of fire acupuncture (FA) on plaque psoriasis (PP), exploring its suitable syndrome types, in order to achieve better therapeutic effects, accelerate the possibility of psoriasis skin lesion recovery, and provide assistance for clinical treatment. METHODS: A total of 8 patients with PP aged between 18 and 60 years were recruited and treated with FA once a week, and the lesion area and severity index (PASI), visual analog scale and pruritus were measured before, 2, 4 and 8 weeks after treatment and at the follow-up period (week 12), respectively. Visual analog scale, and dermoscopy were used for assessment. RESULTS: All patients showed improvement in pruritus after 1 FA treatment, and lesions were reduced to varying degrees after 2 weeks. Except for patients 5 and 8, who only achieved effective results due to severe disease, all other patients with psoriasis achieved significant results at 8 weeks after treatment. CONCLUSION: FA can significantly control the development of lesions, reduce the symptoms of PP lesions and pruritus, and help prevent psoriasis recurrence.


Assuntos
Terapia por Acupuntura , Psoríase , Humanos , Lactente , Psoríase/tratamento farmacológico , Resultado do Tratamento , Prurido/etiologia , Prurido/terapia , Pesquisa , Índice de Gravidade de Doença , Método Duplo-Cego
2.
Zhongguo Zhen Jiu ; 43(3): 255-60, 2023 Mar 12.
Artigo em Chinês | MEDLINE | ID: mdl-36858384

RESUMO

OBJECTIVE: To observe the effect of Shugan Tiaoshen acupuncture (acupuncture for soothing the liver and regulating the mentality) combined with western medication on depression and sleep quality in the patients with depression-insomnia comorbidity due to COVID-19 quarantine, and investigate the potential mechanism from the perspective of cortical excitability. METHODS: Sixty patients with depression-insomnia comorbidity due to COVID-19 quarantine were randomly divided into an acupuncture group and a sham-acupuncture group, 30 cases in each one. The patients of both groups were treated with oral administration of sertraline hydrochloride tablets. In the acupuncture group, Shugan Tiaoshen acupuncture was supplemented. Body acupuncture was applied to Yintang (GV 24+), Baihui (GV 20), Hegu (LI 4), Zhaohai (KI 6), Qihai (CV 6), etc. The intradermal needling was used at Xin (CO15), Gan (CO12) and Shen (CO10). In the sham-acupuncture group, the sham-acupuncture was given at the same points as the acupuncture group. The compensatory treatment was provided at the end of follow-up for the patients in the sham-acupuncture group. In both groups, the treatment was given once every two days, 3 times a week, for consecutive 8 weeks. The self-rating depression scale (SDS) and insomnia severity index (ISI) scores were compared between the two groups before and after treatment and 1 month after the end of treatment (follow-up) separately. The cortical excitability indexes (resting motor threshold [rMT], motor evoked potential amplitude [MEP-A], cortical resting period [CSP]) and the level of serum 5-hydroxytryptamine (5-HT) were measured before and after treatment in the two groups. RESULTS: After treatment and in follow-up, SDS and ISI scores were decreased in both groups compared with those before treatment (P<0.05), and the scores in the acupuncture group were lower than those in the sham-acupuncture group (P<0.05), and the decrease range in the acupuncture group after treatment was larger than that in the sham-acupuncture group (P<0.05). After treatment, rMT was reduced (P<0.05), while MEP-A and CSP were increased (P<0.05) in the acupuncture group compared with that before treatment. The levels of serum 5-HT in both groups were increased compared with those before treatment (P<0.05). The rMT in the acupuncture group was lower than that in the sham-acupuncture group, while MEP-A and CSP, as well as the level of serum 5-HT were higher in the acupuncture group in comparison with the sham-acupuncture group (P<0.05). CONCLUSION: Shugan Tiaoshen acupuncture combined with western medication can relieve depression and improve sleep quality in the patients with depression-insomnia comorbidity due to COVID-19 quarantine, which is probably related to rectifying the imbalanced excitatory and inhibitory neuronal functions.


Assuntos
Terapia por Acupuntura , COVID-19 , Distúrbios do Início e da Manutenção do Sono , Humanos , Depressão , Quarentena , Serotonina , Comorbidade
3.
Front Nutr ; 9: 999189, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36313094

RESUMO

Background: Previous studies showed conflicting evidence on the association between the intake of dietary branched-chain amino acid (BCAA) and the risk of cardiovascular disease (CVD). However, this relationship has not been studied in patients with type 2 diabetes. Therefore, we evaluated the effects of total and individual dietary BCAA (leucine, isoleucine, and valine) intake on CVD risk among individuals with type 2 diabetes in China. Materials and methods: A total of 419 patients with type 2 diabetes who have been diagnosed with CVD (within 2 weeks) were recruited between March 2013 and September 2015 in China. Cases with CVD were 1:1 matched to controls with type 2 diabetes but without CVD by age (±5 years) and sex. A validated 79-item semiquantitative food frequency questionnaire (FFQ) was administered to assess the participants' dietary data. Total dietary BCAA per individual was the summation of the daily intake of isoleucine, leucine, and valine. OR and corresponding CIs were computed by conditional logistic regression models adjusted for potential confounders. Results: Median values of the daily intake of total BCAA were 11.87 g, with an interquartile range of 10.46-13.15 g for cases, and 12.47 g, with an interquartile range of 11.08-13.79 g for controls (P = 0.001). Dietary BCAA was inversely related to CVD risk after multivariable adjustment (OR Q4-Q1 = 0.23, 95%CI = 0.10, 0.51, P trend <0.001 for total BCAA; OR Q4-Q1 = 0.20, 95%CI = 0.07, 0.53, P trend = 0.001 for leucine). For each 1-S.D. increase in total dietary BCAA, leucine or valine intake was associated with 54% (95%CI = 29%, 70%, P = 0.001), 64% (95%CI = 29%, 82%, P = 0.003), or 54% (95%CI = 1%, 79%, P = 0.049) decrease in the risk of CVD, respectively. Whole grains, starchy vegetables, mushrooms, fruit, eggs, and dairy and dairy product-derived BCAA were found to attenuate CVD risk (P ranged: = 0.002-0.027). Conclusion: Higher BCAA intake, in particular leucine and valine, might be associated with a lower risk of CVD.

4.
Infect Immun ; 87(1)2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30348825

RESUMO

Yersinia pseudotuberculosis is a Gram-negative enteropathogen and causes gastrointestinal infections. It disseminates from gut to mesenteric lymph nodes (MLNs), spleen, and liver of infected humans and animals. Although the molecular mechanisms for dissemination and infection are unclear, many Gram-negative enteropathogens presumably invade the small intestine via Peyer's patches to initiate dissemination. In this study, we demonstrate that Y. pseudotuberculosis utilizes its lipopolysaccharide (LPS) core to interact with CD209 receptors, leading to invasion of human dendritic cells (DCs) and murine macrophages. These Y. pseudotuberculosis-CD209 interactions result in bacterial dissemination to MLNs, spleens, and livers of both wild-type and Peyer's patch-deficient mice. The blocking of the Y. pseudotuberculosis-CD209 interactions by expression of O-antigen and with oligosaccharides reduces infectivity. Based on the well-documented studies in which HIV-CD209 interaction leads to viral dissemination, we therefore propose an infection route for Y. pseudotuberculosis where this pathogen, after penetrating the intestinal mucosal membrane, hijacks the Y. pseudotuberculosis-CD209 interaction antigen-presenting cells to reach their target destinations, MLNs, spleens, and livers.


Assuntos
Moléculas de Adesão Celular/metabolismo , Células Dendríticas/microbiologia , Endocitose , Interações Hospedeiro-Patógeno , Lectinas Tipo C/metabolismo , Lipopolissacarídeos/metabolismo , Macrófagos/microbiologia , Receptores de Superfície Celular/metabolismo , Yersinia pseudotuberculosis/patogenicidade , Animais , Aderência Bacteriana , Células Cultivadas , Modelos Animais de Doenças , Humanos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Ligação Proteica , Yersiniose/microbiologia , Yersiniose/patologia , Yersiniose/fisiopatologia
6.
Zhen Ci Yan Jiu ; 43(4): 251-4, 2018 Apr 25.
Artigo em Chinês | MEDLINE | ID: mdl-29888580

RESUMO

OBJECTIVE: To study the clinical efficacy and safety of bee-venom acupuncture therapy for rheumatoid arthritis (RA). METHODS: A total of 120 cases of RA patients were randomized into bee-sting acupuncture group (treatment) and western medicine group (control) in accordance with the random number table. The patients of the control group were treated by oral administration of Methotrexate (10 mg, once a week) and Celecoxlb (0.2 g, once a day), and those of the treatment group treated by 5 to 15 bee stings of Ashi-points or acupoints according to different conditions and corporeity, and with the bee-sting retained for about 5 min every time, once every other day. The treatment lasted for 8 weeks. The therapeutic effect was assessed by examining symptoms and signs of the affected joints as morning stiffness duration, swollen/tender joint counts (indexes), handgrip strength, 15 m-walking time, visual analogue scale (VAS), Disease Activity Score including a 28-joint count (DAS 28), rheumatoid factor (RF), erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), anti-cyclic citrullinated peptide antibody (ACCPA); and for assessing the safety of bee-venom acupuncture, the patients' responses of fever, enlargement of lymph nodes, regional red and swollen, itching, blood and urine tests for routine were examined. RESULTS: Findings of DAS 28 responses displayed that of the two 60 cases in the control and bee-venom acupuncture groups, 15 and 18 experienced marked improvement, 33 and 32 were effective, 12 and 10 ineffective, with the effective rates being 80% and 83. 33%, respectively. No significant difference was found between the two groups in the effective rate (P>0.05). After the treatment, both groups have witnessed a marked decrease in the levels of morning stiffness duration, arthralgia index, swollen joint count index, joint tenderness index, 15 m walking time, VAS, RF, ESR, CRP and ACCPA, and an obvious increase of handgrip strength relevant to their own levels of pre-treatment in each group (P<0.05). There were no significant differences between the two groups in the abovementioned indexes (P>0.05). The routine blood test, routine urine test, routine stool test, electrocardiogram result, the function of liver and kidney and other security index were within the normal range, without any significant adverse effects found after bee-stinging treatment. CONCLUSION: Bee-venom acupuncture therapy for RA patients is safe and effective, worthy of popularization and application in clinical practice.


Assuntos
Terapia por Acupuntura , Artrite Reumatoide , Venenos de Abelha , Animais , Artrite Reumatoide/terapia , Abelhas , Força da Mão , Humanos , Metotrexato , Resultado do Tratamento
7.
Artigo em Inglês | MEDLINE | ID: mdl-28717039

RESUMO

Although the de novo folate biosynthesis pathway has been well studied in bacteria, little is known about its regulation. In the present study, the sigB gene in Mycobacterium tuberculosis was deleted. Subsequent drug susceptibility tests revealed that the M. tuberculosis ΔsigB strain was more sensitive to para-aminosalicylic acid (PAS) and sulfamethoxazole. Comparative transcriptional analysis was performed, and downregulation of pabB was observed in the ΔsigB strain, which was further verified by a quantitative reverse transcription-PCR and Western blot assay. Then, the production levels of para-aminobenzoic acid (pABA) were compared between the sigB deletion mutant and wild-type strain, and the results showed that sigB deletion resulted in decreased production of pABA. In addition, SigB was able to recognize the promoter of pabB in vitro Furthermore, we found that deleting pabC also caused increased susceptibility to PAS. Taken together, our data revealed that, in M. tuberculosis, sigB affects susceptibility to antifolates through multiple ways, primarily by regulating the expression of pabB To our knowledge, this is the first report showing that SigB modulates pABA biosynthesis and thus affecting susceptibility to antifolates, which broadens our understanding of the regulation of bacterial folate metabolism and mechanisms of susceptibility to antifolates.


Assuntos
Ácido 4-Aminobenzoico/metabolismo , Ácido Aminossalicílico/farmacologia , Antituberculosos/farmacologia , Proteínas de Bactérias/genética , Mycobacterium tuberculosis/efeitos dos fármacos , Fator sigma/genética , Sulfametoxazol/farmacologia , Ácido Fólico/metabolismo , Deleção de Genes , Liases/genética , Testes de Sensibilidade Microbiana , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crescimento & desenvolvimento
8.
Histopathology ; 68(3): 378-87, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26040650

RESUMO

AIMS: To study three atypical glioneuronal tumours (GNTs), in order to shed light on the clinical and pathological features of this diverse tumour group. METHODS AND RESULTS: Clinical and neuropathological data for each case were retrospectively reviewed. Case 1 involved a 17-year-old boy with left leg movement difficulty. A mass lesion in the basal ganglia was detected radiologically; histopathological features included neurocytic/perivascular rosettes and a pilocytic astrocytoma component. Case 2 involved a 33-year-old man with intractable epilepsy. His left parietal lobe contained a cyst-like mass, resembling dysembryoplastic neuroepithelial tumour and rosette-forming glioneuronal tumour of the fourth ventricle microscopically. Case 3 involved a 21-year-old woman with a mass lesion in the mesencephalic tegmentum extending to the third and fourth ventricles and the suprasellar region. The lesion contained perivascular/neurocytic rosettes and an oligodendroglioma-like component. None of the tumours expressed an isocitrate dehydrogenase I mutation of the R132H type or contained a 1p/19q deletion, a BRAF(V600E) mutation, or KIAA1549-BRAF fusion. CONCLUSIONS: We describe three GNTs with atypical histopathology and locations. Additional cases and molecular studies are needed to better understand the biological nature of GNTs and to refine their classification system.


Assuntos
Astrocitoma/patologia , Neoplasias Encefálicas/patologia , Glioma/patologia , Neoplasias Neuroepiteliomatosas/patologia , Adolescente , Adulto , Astrocitoma/genética , Gânglios da Base/patologia , Neoplasias Encefálicas/genética , Feminino , Quarto Ventrículo/patologia , Glioma/genética , Humanos , Masculino , Neoplasias Neuroepiteliomatosas/genética , Lobo Parietal/patologia , Estudos Retrospectivos , Formação de Roseta , Adulto Jovem
10.
Neuropathol Appl Neurobiol ; 41(3): 333-46, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24861831

RESUMO

AIMS: Angiocentric glioma (AG) is a rare, slow-growing tumour of the central nervous system. It is often associated with refractory epilepsy and occurs most commonly in children and young adults. We herein report nine cases of AG, including four with atypical histological findings. METHODS: The clinical data and clinicopathological findings of nine cases with AG histological features were described. RESULTS: All nine patients had a history of refractory epilepsy with a mean history of 4.4 years and a median age of 17.6 years at surgery. The AG lesions were located in the superficial cerebrocortical region. Histological examination of these cases revealed characteristic structural features of AG, including bipolar spindle-shaped cells with an angiocentric growth pattern. However, four cases also exhibited atypical histological features: one had astroblastoma-like characteristics, two had a distinct cystic region with an onion-like structure and myxoid changes, and the other one had a region involving many abnormal neurones reminiscent to ganglioglioma. All were positive for glial fibrillary acidic protein and vimentin. Eight cases were positive for epithelial membrane antigen (EMA), with a dot-like staining pattern. A diffuse D2-40 staining was visible in these cases, with two having similar staining pattern to EMA. All cases were immunonegative for BRAF V600E and isocitrate dehydrogenase-1 R132H mutations. CONCLUSIONS: Our results demonstrate that atypical histological features can be present in AG. A collection of more cases and further molecular analyses are required to confirm our findings.


Assuntos
Neoplasias Encefálicas/patologia , Glioma/patologia , Adolescente , Biomarcadores Tumorais/análise , Neoplasias Encefálicas/complicações , Criança , Pré-Escolar , Epilepsia/etiologia , Feminino , Glioma/complicações , Humanos , Imuno-Histoquímica , Imageamento por Ressonância Magnética , Masculino , Adulto Jovem
11.
Brain Pathol ; 25(3): 248-55, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25040707

RESUMO

Sturge-Weber syndrome (SWS) is a rare syndrome characterized by capillary-venous malformations involving skin and brain. Many patients with SWS also suffer from drug-resistant epilepsy. We retrospectively studied a series of six SWS patients with epilepsy and extensive neurosurgical resections. At time of surgery, the patients' age ranged from 11 to 35 years (with a mean of 20.2 years). All surgical specimens were well preserved, which allowed a systematic microscopical inspection utilizing the 2011 ILAE classification for focal cortical dysplasia (FCD). Neuropathology revealed dysmorphic-like neurons with hypertrophic cell bodies reminiscent to those described for FCD type IIa in all cases. However, gross architectural abnormalities of neocortical layering typical for FCD type IIa were missing, and we propose to classify this pattern as FCD ILAE type IIIc. In addition, our patients with earliest seizure onset also showed polymicrogyria (PMG; n = 4). The ictal onset zones were identified in all patients by subdural electrodes, and these areas always showed histopathological evidence for FCD type IIIc. Four out of five patients had favorable seizure control after surgery with a mean follow-up period of 1.7 years. We concluded from our study that FCD type IIIc and PMG are frequently associated findings in SWS. FCD type IIIc may play a major epileptogenic role in SWS and complete resection of the associated FCD should be considered a prognostic key factor to achieve seizure control.


Assuntos
Encéfalo/patologia , Epilepsia/complicações , Epilepsia/cirurgia , Malformações do Desenvolvimento Cortical do Grupo III/complicações , Células Piramidais/patologia , Síndrome de Sturge-Weber/complicações , Adolescente , Adulto , Anticonvulsivantes/uso terapêutico , Encéfalo/cirurgia , Criança , Epilepsia/tratamento farmacológico , Feminino , Humanos , Hipertrofia/complicações , Estudos Longitudinais , Masculino , Malformações do Desenvolvimento Cortical do Grupo III/cirurgia , Estudos Retrospectivos , Síndrome de Sturge-Weber/cirurgia , Adulto Jovem
12.
Zhong Yao Cai ; 34(7): 1055-7, 2011 Jul.
Artigo em Chinês | MEDLINE | ID: mdl-22066397

RESUMO

OBJECTIVE: To investigate the chemical constituents of Carya cathayensis and their antitumor bioactivity. METHODS: The compounds were isolated by Sephadex LH-20 and silica gel column chromatography. Their structures were identified by physicochemical properties and spectroscopic analysis. Then their cytotoxic activity was studied. RESULTS: Five compounds were elucidated as chrysophanol (1), physcion (2), beta-sitosterol (3), pinostrobin(4), 4,8-dihydroxy-1-tetralone (5). CONCLUSION: Compounds 2 and 5 are isolated from Carya cathayensis for the first time. In the MTT antitumor experiments, the compounds 1,4 and 5 have the cytotoxic activity to KB cell.


Assuntos
Carya/química , Medicamentos de Ervas Chinesas/farmacologia , Emodina/análogos & derivados , Tetralonas/isolamento & purificação , Antraquinonas/química , Antraquinonas/isolamento & purificação , Antraquinonas/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Medicamentos de Ervas Chinesas/química , Medicamentos de Ervas Chinesas/isolamento & purificação , Emodina/química , Emodina/isolamento & purificação , Flavanonas/química , Flavanonas/isolamento & purificação , Flavanonas/farmacologia , Frutas/química , Células Hep G2 , Humanos , Espectroscopia de Ressonância Magnética , Casca de Planta/química , Tetralonas/química , Tetralonas/farmacologia
13.
J Biomed Sci ; 18: 43, 2011 Jun 17.
Artigo em Inglês | MEDLINE | ID: mdl-21682866

RESUMO

BACKGROUND: Osteoarthritis (OA) is a degenerative joint disease which affects the entire joint structure, including the synovial membrane. Disease progression was shown to involve inflammatory changes mediated by proteinase-activated receptor (PAR)-2. Previous studies demonstrated that PAR-2 messenger (m)RNA and protein levels increased in OA synovial cells, suggesting that PAR-2 is a potential therapeutic target of the disease. METHODS: We designed a PAR-2-inhibiting peptide (PAR2-IP) by changing an isoleucine residue in the PAR-2-activating peptide (PAR2-AP), SLIGKV, to alanine, generating the SLAGKV peptide. We used it to test PAR-2-mediated inflammatory responses, including the expressions of cyclooxygenase (COX)-2 and matrix metalloproteinase (MMP)-1 and activation of nuclear factor (NF)-κB in human synovial cells. As a control, expressions of COX-2 and MMP-1 were induced by trypsin at both the mRNA and protein levels. RESULTS: The PAR2-AP increased the expression of COX-2 more dramatically than that of MMP-1. When we treated cells with the designed PAR2-IP, the trypsin-induced COX-2 level was completely inhibited at a moderate concentration of the PAR2-IP. With further examination of trypsin-induced NF-κB activation, we observed sufficient inhibitory effects of the PAR2-IP in synoviosarcoma cells and primary synovial cells from OA patients. CONCLUSIONS: Our study suggests that the PAR2-IP inhibits trypsin-induced NF-κB activation, resulting in a reduction in inflammatory COX-2 expression in synovial cells. Application of PAR2-IP is suggested as a potential therapeutic strategy for OA.


Assuntos
Oligopeptídeos/farmacologia , Osteoartrite/tratamento farmacológico , Receptor PAR-2/antagonistas & inibidores , Células Cultivadas , Ciclo-Oxigenase 2/metabolismo , Humanos , Metaloproteinase 1 da Matriz/metabolismo , NF-kappa B/metabolismo , Oligopeptídeos/química , Receptor PAR-2/genética , Receptor PAR-2/metabolismo , Membrana Sinovial/citologia , Membrana Sinovial/metabolismo , Tripsina/metabolismo
14.
Yi Chuan Xue Bao ; 33(12): 1105-11, 2006 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-17185170

RESUMO

Four transgenic soybean lines generated via Agrobacterium-mediated transformation were used to analyze inheritance of the transgenes. Seed chip GUS assay and herbicide leaf painting and spraying assays were applied to test the gus reporter gene and the herbicide resistant bar selectable marker gene, respectively. Three of the four transgenic soybean lines were stably inherited in a Mendelian fashion with co-segregation of both transgenes in a 3:1 segregation ratio in the T(1) progeny, indicating that both transgenes were integrated into the same locus of the soybean genome. Homozygous transgenic progeny plants were obtained in the T(2) generation of these lines, and the transgenes were inherited in five successive generations. However, in one transgenic line, all the T(1) progeny plants showed GUS negative and herbicide sensitive. Southern blotting analysis confirmed that the transgenes were passed into the T(1) progeny, indicating that the transgenes were both silenced. To test if the transgene silencing was due to transcriptional or post-transcriptional level, Soybean mosaic virus (SMV) was inoculated on leaf tissues of the T(1) plants to test possible reverse effects on transgene silencing. Infection with SMV did not suppress transgene silencing, suggesting that transgene silencing in this transgenic line may not be due to post-transcriptional gene silencing.


Assuntos
Glycine max/efeitos dos fármacos , Glycine max/genética , Resistência a Herbicidas/genética , Padrões de Herança/genética , Transgenes/genética , Inativação Gênica , Vírus do Mosaico/fisiologia , Plantas Geneticamente Modificadas , Glycine max/virologia
15.
Sheng Wu Gong Cheng Xue Bao ; 21(6): 970-4, 2005 Nov.
Artigo em Chinês | MEDLINE | ID: mdl-16468355

RESUMO

Two new MAR segments (M14 and M17) were cloned from tobacco genome. Both of the sequences contained several typical consensus sequences of MARs, which were different from the original MAR sequence, such as 90%AT-box, A-box, T-box, the base unpairing regions (BUR), autonomously replicating sequences (ARS), the consensus sequence for topoisomerase II, MAR recognition sequence (MRS), origin of replication (ORI), curved DNA motifs and ATATTT et al. To investigate the effects of these two sequences on gene expression in transgenic plants, 3 plant expression vectors were constructed with uidA gene coding beta-glucuronidase (GUS) which were flanked on one side and on both sides by the MARs we obtained. These plant expression vectors with one or two MARs were transformed into tobaccos via Agrobacterium-mediated transformation method, with the plant expression vector pCAMBIA2301 without MAR and wild type tobacco as controls. GUS histochemical staining results showed that the uidA gene expressed stably in transgenic tobaccos. Quantitative detection of GUS activity showed that the MARs could increase GUS expression levels in vivo in contrast to the controls, wherever they were flanked on one side or both sides of uidA gene. The vector ligated with MARs in the same direction on both sides of uidA could increase the GUS expression level much better than both vectors which just ligated with single MARs on one side. The former one increased the average GUS activity for 3.14 folds, but 1.56 and 2.43 folds for the latter two vectors with single MARs respectively contrasting to the pCAMBIA2301 control. But the expression differences among individual transformants were still obvious. Therefore, it was concluded that the DNA sequences we obtained in this experiment were two novel MARs and could enhance gene expression in vivo. In the meanwhile, although the numbers of the MARs typical motifs in M14 were more than in M17, especially the 90% AT box which had been considered to be the highest correlative motif with binding strength in vitro, the enhancement of gene expression was lower yet, which implied no correlation between improvement of gene expression and binding strength between MARs and nuclear matrix in vitro.


Assuntos
DNA de Plantas/genética , Regiões de Interação com a Matriz , Nicotiana/genética , Matriz Nuclear/metabolismo , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Regulação da Expressão Gênica de Plantas , Vetores Genéticos , Plantas Geneticamente Modificadas/genética , Análise de Sequência de DNA
16.
Sheng Wu Gong Cheng Xue Bao ; 20(5): 646-51, 2004 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-15973983

RESUMO

Because of the concern about escape of antibiotic- or herbicide-resistant transgenes from transgenic crops, selectable marker genes from plant origin would be an alternative choice for plant transformation. In this study, a feedback-insensitive anthranilate synthase gene ( ASA2 ) cloned from a tobacco cell line was tested for Agrobacterium-mediated transformation of axis tissue of soybean mature embryo, with a tryptophan analogue 5-methyltryptophan (5-MT) as the selective agent. Southern blot analysis of the To transgenic lines confirmed the integration of the ASA2 gene into the soybean genome. Northern blot analysis showed the ASA2 gene was also expressed in the leave tissue, and the free tryptophan content in the leaf tissue of transgenic soybean was about 59% to 123% more than that in the wild type. PCR analysis of the T1 progeny showed that the transgene was inherited in a Mendelian fashion. All these results indicate that this feedback-insensitive ASA2 gene can be used as a selectable marker gene for plant transformation. This work also demonstrated that the ASA2 gene coding for the a-subunits from one plant (tobacco) can interact with the n-subunits of a heterologous plant (soybean) to form an active anthranilate synthase enzyme. The use of this feedback-insensitive gene as a novel selectable marker for plant transformation is also discussed.


Assuntos
Antranilato Sintase/genética , Glycine max/genética , Plantas Geneticamente Modificadas/genética , Transformação Genética , Retroalimentação Fisiológica , Triptofano/análogos & derivados , Triptofano/metabolismo
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