RESUMO
Photoperiod/temperature-sensitive genic male sterility (P/TGMS) is widely applied for improving crop production. Previous investigations using the reversible male sterile (rvms) mutant showed that slow development is a general mechanism for restoring fertility to P/TGMS lines in Arabidopsis. In this work, we isolated a restorer of rvms-2 (res3), as the male sterility of rvms-2 was rescued by res3. Phenotype analysis and molecular cloning show that a point mutation in UPEX1 l in res3 leads to delayed secretion of callase A6 from the tapetum to the locule and tetrad callose wall degradation. Electrophoretic mobility shift assay and chromatin immunoprecipitation analysis demonstrated that the tapetal transcription factor ABORTED MICROSPORES directly regulates UPEX1 expression, revealing a pathway for tapetum secretory function. Early degradation of the callose wall in the transgenic line eliminated the fertility restoration effect of res3. The fertility of multiple known P/TGMS lines with pollen wall defects was also restored by res3. We propose that the remnant callose wall may broadly compensate for the pollen wall defects of P/TGMS lines by providing protection for pollen formation. A cellular mechanism is proposed to explain how slow development restores the fertility of P/TGMS lines in Arabidopsis.
Assuntos
Proteínas de Arabidopsis , Arabidopsis , Infertilidade Masculina , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fertilidade/genética , Glucanos , Infertilidade Masculina/metabolismo , Fotoperíodo , Infertilidade das Plantas/genética , Pólen/metabolismo , TemperaturaRESUMO
The objective of this study was to compare the efficacy and safety of 10 different surgical treatments for benign prostatic hyperplasia (BPH) with volume >60 mL. A systematic literature review and network meta-analysis of randomized controlled trials (RCTs) within a Bayesian framework was performed. A total of 52 parallel-group RCTs included, reporting on 6,947 participants, comparing open prostatectomy (OP), monopolar/bipolar transurethral resection of prostate (monopolar/ bipolar TURP), thulium, holmium and diode laser enucleation of prostate (LEP), bipolar enucleation of prostate, potassium titanyl phosphate laser vaporization of prostate (KTP LVP), bipolar vaporization of prostate (bipolar VP), and laparoscopic simple prostatectomy (laparoscope SP). Compared with OP, laparoscope SP identified better maximal flow rate (Qmax; mean differences [MDs] = 2.89 mL/s) at the 24th month, but bipolar VP demonstrated worse Qmax (MD = -3.20 mL/s) and International Prostate Symptom Score (IPSS; MD = 2.60) at the 12th month. Holmium LEP (MD = 1.37) demonstrated better International Index of Erectile Function-5 at the 12th month compared with OP. However, compared with OP, KTP LVP demonstrated worse postvoid residual volume (PVR) at the sixth (MD = 10.42 mL) and 12th month (MD = 5.89 mL) and monopolar TURP (MD = 6.9 mL) demonstrated worse PVR at the 12th month. Eight new surgical methods for BPH with volume >60 mL appeared to be superior in safety compared with OP and monopolar TURP due to fewer complications. Bipolar VP and KTP LVP maybe not suitable for prostates more than 60 mL due to short- and middle-term worse Qmax, IPSS, and PVR than OP.
Assuntos
Hiperplasia Prostática , Ressecção Transuretral da Próstata , Humanos , Masculino , Metanálise em Rede , Hiperplasia Prostática/cirurgia , Qualidade de Vida , Ensaios Clínicos Controlados Aleatórios como Assunto , Resultado do TratamentoRESUMO
Severe Coronavirus Disease 2019 (COVID-19) is characterized by numerous complications, complex disease, and high mortality, making its treatment a top priority in the treatment of COVID-19. Integrated traditional Chinese medicine (TCM) and western medicine played an important role in the prevention, treatment, and rehabilitation of COVID-19 during the epidemic. However, currently there are no evidence-based guidelines for the integrated treatment of severe COVID-19 with TCM and western medicine. Therefore, it is important to develop an evidence-based guideline on the treatment of severe COVID-19 with integrated TCM and western medicine, in order to provide clinical guidance and decision basis for healthcare professionals, public health personnel, and scientific researchers involved in the diagnosis, treatment, and care of COVID-19 patients. We developed and completed the guideline by referring to the standardization process of the "WHO handbook for guideline development", the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system, and the Reporting Items for Practice Guidelines in Healthcare (RIGHT).
Assuntos
Antivirais/uso terapêutico , Tratamento Farmacológico da COVID-19 , Medicamentos de Ervas Chinesas/uso terapêutico , Infectologia/tendências , Medicina Tradicional Chinesa/tendências , SARS-CoV-2/efeitos dos fármacos , Antivirais/efeitos adversos , COVID-19/diagnóstico , COVID-19/virologia , Consenso , Técnica Delphi , Medicamentos de Ervas Chinesas/efeitos adversos , Medicina Baseada em Evidências/tendências , Interações Hospedeiro-Patógeno , Humanos , Gravidade do Paciente , SARS-CoV-2/patogenicidade , Resultado do TratamentoRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Description of the pharmacological activities of Sanghuang mushrooms (Inonotus Sanghuang) can be traced back to Tang dynasty of China 1300 years ago. This mushroom has been widely accepted in China, Japan, Korea and certain regions of Europe as a nutraceutical medicine for enhancing immunity or an alternative medicine for prevention or inhibition of tumorigenesis. However, this mushroom is rarely available from the mulberry trees in the wild because of the rigorous conditions needed for formation of the Sanghuang mushrooms. AIM OF THE STUDY: This study aims to establish a practical protocol for culture, particularly for a bunch of production of Sanghuang mushrooms possibly to commercialize the cultured Sanghuang based on deep comparison of quality and pharmacological activities between the cultured and the wild Sanghuang. MATERIALS AND METHODS: A phylogenetic tree containing five strains of the wild Sanghuang was constructed using rDNA markers. Different temperatures and medium compositions were surveyed to develop a practical protocol for culture of the Sanghuang mushrooms. 5-fluorouracil was used to induce the immunodeficient mice. Chemotherapeutic components and pharmacological activities were deeply analyzed between a cultured strain (SG) and three strains of the wild Sanghuang. RESULTS: Maintenance of a temperature of 22-28⯰C and a high relative humidity of 90-95%, and use of a high ratio (80%) of mulberry tree sticks in the medium were critical to successful culture of Sanghuang. The cultured mushrooms were yellow with a uniform shape, while the wild Sanghuang was dark brown with a smaller and irregular shape. The cultured mushrooms contained significantly higher levels of polysaccharides, amino acids, and water-soluble nutraceuticals, whereas flavones in the wild Sanghuang were significantly higher (Pâ¯<â¯0.05). Use of a dose of 8â¯mg/kg or 16â¯mg/kg to immunoregenerate the immunodeficient mice was comparable between the cultured and wild Sanghang based on analysis of hematological parameters and histological examination of the thymus and spleen in the treated mice. CONCLUSIONS: This study highlights the potential of the immunoregenerative functions of the cultured Sanghuang for cancer chemotherapy and suggests that the cultured Sanghuang can be an alternative to wild Sanghuang used for nutraceutical medicine.
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Agaricales , Síndromes de Imunodeficiência/terapia , Agaricales/genética , Animais , Antineoplásicos , Biônica , Contagem de Células Sanguíneas , Feminino , Fluoruracila , Síndromes de Imunodeficiência/induzido quimicamente , Camundongos Endogâmicos BALB C , Filogenia , Baço/efeitos dos fármacos , Baço/patologia , Timo/efeitos dos fármacos , Timo/patologiaRESUMO
OBJECTIVE: To investigate the anti-leukemia effect of total saponins of Rubus parvifolius L. (TSRP) on K562 cell xenografts in nude mice and the mechanisms of action. METHODS: The K562 cell xenografts in nude mice were established, and then randomly divided into 5 groups, the control group, the cytosine arabinoside group(Ara-c) and 3 TSRP groups (20, 40 and 100 mg/kg). The tumor volume and mass of each group of nude mice were measured and the anti-tumor rates of TSRP were calculated subsequently. The apoptosis status of tumor cells was detected by hematoxylin-eosin (HE) and terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining analysis. Finally, the activities of apoptosis related signaling of signal transducer and activator of transcription 3 (STAT3), eukaryotic initiation factor 4E (eIF4E) and B-cell lymphoma-2 (bcl-2) were determined with immunohistochemistry tests. RESULTS: Subcutaneous injection of K562 cells induced tumor formation in nude mice, and the TSRP treated group showed a signifificant inhibitory effect on tumor formation. The nude mice treated with TSRP showed a signifificant decrease in tumor growth rate and tumor weight in comparison to the control group (all P<0.05). The HE staining and TUNEL assay showed that TSRP induced cell death by apoptosis. The immunohistochemical assay showed down-regulation of the bcl-2 gene in the TSRP treated cells. The phosphorylation levels of eIF4E and STAT3 were decreased obviously after the treatment of TSRP. CONCLUSION: TSRP had an excellent tumor-suppressing effect on K562 cells in the nude mice xenograft model, suggesting that TSPR can be developed as a promising anti-chronic myeloide leukemia drug.
Assuntos
Fator de Iniciação 4E em Eucariotos/fisiologia , Leucemia/tratamento farmacológico , Rubus , Fator de Transcrição STAT3/fisiologia , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Humanos , Células K562 , Leucemia/patologia , Masculino , Camundongos , Rubus/química , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
BACKGROUND: In Arabidopsis, the tapetum and microsporocytes are critical for pollen formation. Previous studies have shown that ARF17 is expressed in microsporocytes and tetrads and directly regulates tetrad wall synthesis for pollen formation. ARF17 is the direct target of miR160, and promoterARF17::5mARF17 (5mARF17/WT) transgenic plants, which have five silent mutations within the miR160-complementary domain, are sterile. RESULTS: Here, we found that ARF17 is also expressed in the tapetum, which was defective in arf17 mutants. Compared with arf17 mutants, 5mARF17/WT plants had abnormal tapetal cells and tetrads but were less vacuolated in the tapetum. Immunocytochemical assays showed that the ARF17 protein over-accumulated in tapetum, microsporocytes and tetrads of 5mARF17/WT plants at early anther stages, but its expression pattern was not affected during anther development. 5mARF17 driven by its native promoter did not rescue the arf17 male-sterile phenotype. The expression of 5mARF17 driven by the tapetum-specific promoter A9 led to a defective tapetum and male sterility in transgenic plants. These results suggest that the overexpression of ARF17 in the tapetum and microsporocytes of 5mARF17/WT plants leads to male sterility. Microarray data revealed that an abundance of genes involved in transcription and translation are ectopically expressed in 5mARF17/WT plants. CONCLUSIONS: Our work shows that ARF17 plays an essential role in anther development and pollen formation, and ARF17 expression under miR160 regulation is critical for its function during anther development.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/genética , Flores/crescimento & desenvolvimento , Regulação da Expressão Gênica de Plantas , Pólen/crescimento & desenvolvimento , Fatores de Transcrição/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Flores/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/metabolismo , Pólen/genética , Fatores de Transcrição/metabolismoRESUMO
The aberrant alterations of calmodulin (CaM) and its downstream substrates have been reported in some neurodegenerative diseases, but rarely described in prion disease. In this study, the potential changes of Ca2+/CaM and its associated agents in the brains of scrapie agent 263K-infected hamsters and the prion infected cell line SMB-S15 were evaluated by various methodologies. We found that the level of CaM in the brains of 263K-infected hamsters started to increase at early stage and maintained at high level till terminal stage. The increased CaM mainly accumulated in the regions of cortex, thalamus and cerebellum of 263K-infected hamsters and well localization of CaM with NeuN positive cells. However, the related kinases such as total and phosphorylated forms of CaMKII and CaMKIV, as well as the downstream proteins such as CREB and BDNF in the brain of 263K-infected hamsters were decreased. Further analysis showed a remarkable increase of S-nitrosylated (SNO) form of CaM in the brains of 263K-infected hamsters. Dynamic analysis of S-nitrosylated CaM showed the SNO form of CaM abnormally increases in a time-dependent manner during prion infection. Compared with that of the normal partner cell line SMB-PS, the CaM level in SMB-S15 cells was increased, meanwhile, the downstream proteins, such as CaMKII, p-CaMKII, CREB, as well as BDNF, were also increased, especially in the nucleic fraction. No SNO-CaM was detected in the cell lines SMB-S15 and SMB-PS. Our data indicate an aberrant increase of CaM during prion infection in vivo and in vitro.
Assuntos
Fator Neurotrófico Derivado do Encéfalo/metabolismo , Proteína de Ligação a CREB/metabolismo , Calmodulina/metabolismo , Córtex Cerebral/metabolismo , Scrapie/metabolismo , Tálamo/metabolismo , Animais , Cálcio/metabolismo , Proteína Quinase Tipo 2 Dependente de Cálcio-Calmodulina/metabolismo , Proteína Quinase Tipo 4 Dependente de Cálcio-Calmodulina/metabolismo , Linhagem Celular , Córtex Cerebral/patologia , Cricetinae , Modelos Animais de Doenças , Camundongos , Proteínas PrPSc/metabolismo , Tálamo/patologia , Fatores de TempoRESUMO
α1-Antichymotrypsin (α1-ACT) belongs to a kind of acute-phase inflammatory protein. Recently, such protein has been proved exist in the amyloid deposits which is the hallmark of Alzheimer's disease, but limitedly reported in prion disease. To estimate the change of α1-ACT during prion infection, the levels of α1-ACT in the brain tissues of scrapie agents 263K-, 139A- and ME7-infected rodents were analyzed, respectively. Results shown that α1-ACT levels were significantly increased in the brain tissues of the three kinds of scrapie-infected rodents, displaying a time-dependent manner during prion infection. Immunohistochemistry assays revealed the increased α1-ACT mainly accumulated in some cerebral regions of rodents infected with prion, such as cortex, thalamus and cerebellum. Immunofluorescent assays illustrated ubiquitously localization of α1-ACT with GFAP positive astrocytes, Iba1-positive microglia and NeuN-positive neurons. Moreover, double-stained immunofluorescent assays and immunohistochemistry assays using series of brain slices demonstrated close morphological colocalization of α1-ACT signals with that of PrP and PrPSc in the brain slices of 263K-infected hamster. However, co-immunoprecipitation does not identify any detectable molecular interaction between the endogenous α1-ACT and PrP either in the brain homogenates of 263K-infected hamsters or in the lysates of prion-infected cultured cells. Our data here imply that brain α1-ACT is increased abnormally in various scrapie-infected rodent models. Direct molecular interaction between α1-ACT and PrP seems not to be essential for the morphological colocalization of those two proteins in the brain tissues of prion infection.
Assuntos
Córtex Cerebelar/metabolismo , Proteínas PrPSc/metabolismo , Doenças Priônicas/metabolismo , Tálamo/metabolismo , alfa 1-Antiquimotripsina/metabolismo , Amiloide/metabolismo , Animais , Astrócitos/metabolismo , Linhagem Celular , Córtex Cerebelar/patologia , Cricetinae , Proteínas de Ligação a DNA , Modelos Animais de Doenças , Proteína Glial Fibrilar Ácida/metabolismo , Camundongos , Microglia/metabolismo , Proteínas do Tecido Nervoso/metabolismo , Neurônios/metabolismo , Proteínas Nucleares/metabolismo , Proteínas Priônicas/metabolismo , Tálamo/patologia , Fatores de Tempo , alfa 1-Antiquimotripsina/análiseRESUMO
Importance: Acupuncture is used to induce ovulation in some women with polycystic ovary syndrome, without supporting clinical evidence. Objective: To assess whether active acupuncture, either alone or combined with clomiphene, increases the likelihood of live births among women with polycystic ovary syndrome. Design, Setting, and Participants: A double-blind (clomiphene vs placebo), single-blind (active vs control acupuncture) factorial trial was conducted at 21 sites (27 hospitals) in mainland China between July 6, 2012, and November 18, 2014, with 10 months of pregnancy follow-up until October 7, 2015. Chinese women with polycystic ovary syndrome were randomized in a 1:1:1:1 ratio to 4 groups. Interventions: Active or control acupuncture administered twice a week for 30 minutes per treatment and clomiphene or placebo administered for 5 days per cycle, for up to 4 cycles. The active acupuncture group received deep needle insertion with combined manual and low-frequency electrical stimulation; the control acupuncture group received superficial needle insertion, no manual stimulation, and mock electricity. Main Outcomes and Measures: The primary outcome was live birth. Secondary outcomes included adverse events. Results: Among the 1000 randomized women (mean [SD] age, 27.9 [3.3] years; mean [SD] body mass index, 24.2 [4.3]), 250 were randomized to each group; a total of 926 women (92.6%) completed the trial. Live births occurred in 69 of 235 women (29.4%) in the active acupuncture plus clomiphene group, 66 of 236 (28.0%) in the control acupuncture plus clomiphene group, 31 of 223 (13.9%) in the active acupuncture plus placebo group, and 39 of 232 (16.8%) in the control acupuncture plus placebo group. There was no significant interaction between active acupuncture and clomiphene (P = .39), so main effects were evaluated. The live birth rate was significantly higher in the women treated with clomiphene than with placebo (135 of 471 [28.7%] vs 70 of 455 [15.4%], respectively; difference, 13.3%; 95% CI, 8.0% to 18.5%) and not significantly different between women treated with active vs control acupuncture (100 of 458 [21.8%] vs 105 of 468 [22.4%], respectively; difference, -0.6%; 95% CI, -5.9% to 4.7%). Diarrhea and bruising were more common in patients receiving active acupuncture than control acupuncture (diarrhea: 25 of 500 [5.0%] vs 8 of 500 [1.6%], respectively; difference, 3.4%; 95% CI, 1.2% to 5.6%; bruising: 37 of 500 [7.4%] vs 9 of 500 [1.8%], respectively; difference, 5.6%; 95% CI, 3.0% to 8.2%). Conclusions and Relevance: Among Chinese women with polycystic ovary syndrome, the use of acupuncture with or without clomiphene, compared with control acupuncture and placebo, did not increase live births. This finding does not support acupuncture as an infertility treatment in such women. Trial Registration: clinicaltrials.gov Identifier: NCT01573858.
Assuntos
Terapia por Acupuntura , Clomifeno/uso terapêutico , Fármacos para a Fertilidade Feminina/uso terapêutico , Infertilidade Feminina/terapia , Nascido Vivo/epidemiologia , Síndrome do Ovário Policístico/terapia , Terapia por Acupuntura/efeitos adversos , Terapia por Acupuntura/estatística & dados numéricos , Adulto , Índice de Massa Corporal , Clomifeno/efeitos adversos , Terapia Combinada/métodos , Contusões/etiologia , Diarreia/etiologia , Método Duplo-Cego , Esquema de Medicação , Feminino , Fármacos para a Fertilidade Feminina/efeitos adversos , Humanos , Infertilidade Feminina/tratamento farmacológico , Infertilidade Feminina/etiologia , Síndrome do Ovário Policístico/complicações , Síndrome do Ovário Policístico/tratamento farmacológico , Gravidez , Método Simples-Cego , Fatores de TempoRESUMO
OBJECTIVE: To study the therapeutic effect of Lamiophlomis Rotata Capsule (LRC) in the treatment of type â ¢B prostatitis. METHODS: We randomly divided 225 patients with type â ¢B prostatitis into an experimental group (n =125) and a control group (n =120), the former treated orally with LRC at 3 capsules tid while the latter with tamsulosin hydrochloride sustained-release capsules at 0.2 mg qd, both for 4 weeks. We compared the therapeutic effects between the two groups of patients based on the National Institutes of Health Chronic Prostatitis Symptom Index (NIH-CPSI) obtained before, immediately after and at 4 weeks after medication. RESULTS: A total of 120 patients completed the treatment in the experimental group, which showed remarkable decreases as compared with the baseline in the pain score (5.30 ± 1.23 vs 14.68 ± 1.51, P<0.05), quality of life (QoL) score (6.46 ± 0.93 vs 8.52 ± 1.05, P<0.05) and total NIH-CPSI score (17.50 ± 2.77 vs 27.99 ± 2.98, P<0.05) after 4 weeks of treatment, but no significant change in the urination symptoms score (7.41 ± 1.16 vs 7.16 ± 1.04, P>0.05). The experimental group achieved even markedly lower scores than the controls in the pain symptom (5.30 ± 1.23 vs 13.67 ± 1.49, P<0.05), QoL (6.46 ± 0.93 vs 7.47 ± 0.88, P<0.05) and total NIH-CPSI (17.50 ± 2.77 vs 25.77 ± 2.01, P<0.05) but a higher urination symptoms score than the latter after medication (7.16 ± 1.04 vs 5.68 ± 1.34, P<0.05). At 4 weeks after drug withdrawal, the experimental group also showed significantly lower scores of the pain symptom (7.23 ± 1.03), QoL (6.58 ± 0.87) and total NIH-CPSI (22.18 ± 2.03) than the baseline (all P<0.05) and those in the control group (14.14 ± 0.98, 8.12 ± 0.72 and 26.89 ± 1.67) (all P<0.05). Apart from dizziness in 2 of the patients, who gave up medication halfway, no other obvious adverse reactions were observed during the experiment. CONCLUSIONS: Lamiophlomis Rotata Capsule deserves to be recommended for the treatment of type â ¢B prostatitis for its safety and effectiveness in reducing the pain and improving the life quality of the patients.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Prostatite/tratamento farmacológico , Tansulosina/uso terapêutico , Agentes Urológicos/uso terapêutico , Cápsulas , Doença Crônica , Preparações de Ação Retardada , Humanos , Masculino , Manejo da Dor , Prostatite/patologia , Qualidade de Vida , MicçãoRESUMO
BACKGROUND: Genetic diseases represent a significant public health challenge in China that will need to be addressed by a correspondingly large number of professional genetic counselors. However, neither an official training program for genetic counseling, nor formal board certification, was available in China before 2015. In 2009, a genetic counseling training program based on role-playing was implemented as a pilot study at the Third Military Medical University to train third-year medical students. METHODS: Questionnaires on participant attitudes to the program and role-playing were randomly administered to 324 students after they had finished their training. Pre- and post-training instructional tests, focusing on 42 key components of genetic counseling, were administered randomly to 200 participants to assess mastery of each component. Finally, scores in final examinations of 578 participants from 2009 to 2011 were compared to scores obtained by 614 non-participating students from 2006 to 2008 to further assess program efficacy. RESULTS: Both the training program and the instructional strategy of role-playing were accepted by most participants. Students believed that role-playing improved their practice of genetic counseling and medical genetics, enhanced their communication skills, and would likely contribute to future professional performance. The average understanding of 40 of the key points in genetic counseling was significantly improved, and most students approached excellent levels of mastery. Scores in final examinations and the percentages of students scoring above 90 were also significantly elevated. CONCLUSIONS: Role-playing is a feasible and effective instructional strategy for training genetic counselors in China as well as in other developing countries.
Assuntos
Aconselhamento/educação , Educação de Graduação em Medicina/métodos , Aconselhamento Genético , Desempenho de Papéis , Estudantes de Medicina , China/epidemiologia , Aconselhamento/normas , Educação de Graduação em Medicina/normas , Avaliação Educacional , Humanos , Projetos Piloto , Competência Profissional , Avaliação de Programas e Projetos de SaúdeRESUMO
Nexine is a conserved layer of the pollen wall. We previously reported that the nexine layer is absent in the knockout mutant of Arabidopsis TRANSPOSABLE ELEMENT SILENCING VIA AT-HOOK (TEK) gene. In this study, we investigated the molecular regulatory functions of TEK in pollen development and identified the genes encoding Arabinogalactan proteins (AGPs) as direct targets of TEK, which are essential for nexine formation. Phenotypic similarity between tek and the TEK-SRDX transgenic lines suggest that TEK plays a role in transcriptional activation in anther development. Microarray analysis identified a total of 661 genes downregulated in tek, including four genes encoding AGPs, AGP6, AGP11, AGP23, and AGP40. Electrophoretic mobility shift assays showed that TEK could directly bind the nuclear matrix attachment region (MAR) and the promoter of AGP6. Chromatin immunoprecipitation followed by PCR analysis demonstrated that TEK is enriched in the promoters of the four AGP genes. Expression of AGP6 driven by the TEK promoter in tek partially rescued both nexine formation and plant fertility. These results indicate that TEK directly regulates AGP expression in the anther to control nexine layer formation. We also proposed that glycoproteins might be essential components of the nexine layer in the pollen wall.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Parede Celular/metabolismo , Mucoproteínas/metabolismo , Pólen/metabolismo , Regulação da Expressão Gênica de Plantas/fisiologia , Proteínas de Plantas/metabolismo , Regiões Promotoras Genéticas/genéticaRESUMO
The tapetum plays a critical role during the development and maturation of microspores. DYSFUNCTIONAL TAPETUM 1 (DYT1) is essential for early tapetal development. Here, we determined that the promoter region (-550 to -463 bp) contains indispensable cis-elements for DYT1 expression. Although DYT1 transcripts can be detected in both meiocytes and tapetal cells, localization of DYT1-GFP demonstrated that DYT1 is strictly located in tapetal cells during microsporogenesis. Chromatin immunoprecipitation (ChIP) analysis revealed that DYT1 directly binds the promoter region of Defective in Tapetal Development and Function 1 (TDF1), a transcription factor essential for tapetum development. When TDF1 driven by the DYT1 promoter is expressed in a dyt1 mutant, the expression of the transcription factors AMS, MS188/MYB80, TEK and MS1 and the pollen wall-related genes are restored. Although the pollen wall is not formed and the microspores are ruptured, DIOC2 staining showed that fatty acids, the precursors of the pollen wall, were synthesized in the transgenic lines. These results indicate that DYT1 regulates the expression of AMS, MS188/MYB80, TEK and MS1 for pollen wall formation, primarily via TDF1.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Regulação da Expressão Gênica de Plantas , Fatores de Transcrição/genética , Arabidopsis/crescimento & desenvolvimento , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Parede Celular/metabolismo , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Mutação , Plantas Geneticamente Modificadas , Pólen/genética , Pólen/crescimento & desenvolvimento , Pólen/metabolismo , Regiões Promotoras Genéticas , Fatores de Transcrição/metabolismoRESUMO
BACKGROUND: Saponins are a major active component for the traditional Chinese medicine, Rubus parvifolius L., which has shown clear antitumor activities. However, the specific effects and mechanisms of saponins of Rubus parvifolius L. (SRP) remain unclear with regard to human chronic myeloid leukemia cells. The aim of this study was to investigate inhibition of proliferation and apoptosis induction effects of SRP in K562 cells and further elucidate its regulatory mechanisms. MATERIALS AND METHODS: K562 cells were treated with different concentrations of SRP and MTT assays were performed to determine cell viability. Apoptosis induction by SRP was determined with FACS and DAPI staining analysis. Western blotting was used to detect expression of apoptosis and survival related genes. Specific inhibitors were added to confirm roles of STAT3 and AMPK pathways in SRP induction of apoptosis. RESULTS: Our results indicated that SRP exhibited obvious inhibitory effects on the growth of K562 cells, and significantly induced apoptosis. Cleavage of pro-apoptotic proteins was dramatically increased after SRP exposure. SRP treatment also increased the activities of AMPK and JNK pathways, and inhibited the phosphorylation expression level of STAT3 in K562 cells. Inhibition of the AMPK pathway blocked the activation of JNK by SRP, indicating that SRP regulated the expression of JNK dependent on the AMPK pathway. Furthermore, inhibition of the latter significantly conferred resistance to SRP pro- apoptotic activity, suggesting involvement of the AMPK pathway in induction of apoptosis. Pretreatment with a STAT3 inhibitor also augmented SRP induced growth inhibition and cell apoptosis, further confirming roles of the STAT3 pathway after SRP treatment. CONCLUSIONS: Our results demonstrated that SRP induce cell apoptosis through AMPK activation and STAT3 inhibition in K562 cells. This suggests the possibility of further developing SRP as an alternative treatment option, or perhaps using it as adjuvant chemotherapeutic agent for chronic myeloid leukemia therapy.
Assuntos
Apoptose/efeitos dos fármacos , Leucemia Mielogênica Crônica BCR-ABL Positiva/tratamento farmacológico , Rubus/química , Saponinas/farmacologia , Proteínas Quinases Ativadas por AMP/metabolismo , Antineoplásicos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Hormônio Liberador da Corticotropina/metabolismo , Humanos , Células K562 , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-jun/metabolismo , Fator de Transcrição STAT3/metabolismo , Transdução de Sinais/efeitos dos fármacos , Urocortinas/metabolismoRESUMO
The pollen wall, an essential structure for pollen function, consists of two layers, an inner intine and an outer exine. The latter is further divided into sexine and nexine. Many genes involved in sexine development have been reported, in which the MYB transcription factor Male Sterile 188 (MS188) specifies sexine in Arabidopsis. However, nexine formation remains poorly understood. Here we report the knockout of TRANSPOSABLE ELEMENT SILENCING VIA AT-HOOK (TEK) leads to nexine absence in Arabidopsis. TEK encodes an AT-hook nuclear localized family protein highly expressed in tapetum during the tetrad stage. Absence of nexine in tek disrupts the deposition of intine without affecting sexine formation. We find that ABORTED MICROSPORES directly regulates the expression of TEK and MS188 in tapetum for the nexine and sexine formation, respectively. Our data show that a transcriptional cascade in the tapetum specifies the development of pollen wall.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética , Pólen/genética , Sequência de Aminoácidos , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição Hélice-Alça-Hélice Básicos/metabolismo , Parede Celular/fisiologia , Flores/genética , Regulação da Expressão Gênica de Plantas , Pólen/metabolismo , Pólen/fisiologiaRESUMO
OBJECTIVE: To determine the antiproliferative activity of Rubus parvifolius L. (RP) extract, its medicinal serum and RP total saponins (RPTS) against K562 cells in vitro and in vivo. METHODS: Nude mice models bearing leukemia tumors were treated with different concentrations of RP extract. The size, weight and histopathological change of leukemic tumors were determined. Semi-solid agar culture and methylthiazolyl tetrazolium (MTT) assay were used to determine in vitro the inhibition of colony formation and proliferation of K562 cells respectively by different concentrations of RP medicinal serum and RPTS. RESULTS: RP extract had a tumor inhibition rate of 84.8% when administered to mice at a dose of 1.0 g/day of crude RP root equivalent. Semi-solid agar culture of K562 cells in the presence of 20% (v/v) of RP medicinal serum and 150 mg/L RPTS demonstrated a 50.8% and 100% inhibition of the colony forming unit (CFU)-K562, respectively. The same doses of RP medicinal serum and RPTS showed a proliferation inhibition of 31.4% and 86.3%, respectively against K562 cells in MTT assay. CONCLUSION: RP extract and RPTS show effective antiproliferative activity against myeloid leukemia cells in vitro and in vivo.
Assuntos
Leucemia/patologia , Extratos Vegetais/farmacologia , Rosaceae/química , Ágar , Animais , Proliferação de Células/efeitos dos fármacos , Cromatografia Líquida de Alta Pressão , Humanos , Células K562 , Leucemia/tratamento farmacológico , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Extratos Vegetais/uso terapêutico , Saponinas/farmacologia , Saponinas/uso terapêutico , Tela Subcutânea/patologia , Ensaio Tumoral de Célula-Tronco , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The adaptations of root morphology, physiology, and biochemistry to phosphorus supply have been characterized intensively. However, characterizing these adaptations at molecular level is largely neglected under field conditions. Here, two consecutive field experiments were carried out to investigate the agronomic traits and root traits of wheat (Triticum aestivum L.) at six P-fertilizer rates. Root samples were collected at flowering to investigate root dry weight, root length density, arbusular-mycorrhizal colonization rate, acid phosphatase activity in rhizosphere soil, and expression levels of genes encoding phosphate transporter, phosphatase, ribonucleases, and expansin. These root traits exhibited inducible, inhibitory, or combined responses to P deficiency, and the change point for responses to P supply was at or near the optimal P supply for maximum grain yield. This research improves the understanding of mechanisms of plant adaptation to soil P in intensive agriculture and provides useful information for optimizing P management based on the interactions between soil P dynamics and root processes.
Assuntos
Fósforo/farmacologia , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/genética , Triticum/anatomia & histologia , Triticum/genética , Biomassa , Flores/efeitos dos fármacos , Flores/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas/genética , Fósforo/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/genética , Brotos de Planta/metabolismo , Solo , Triticum/efeitos dos fármacos , Triticum/crescimento & desenvolvimentoRESUMO
OBJECTIVE: To establish HPLC fingerprint analysis methods of anthraquinone components of Morinda officinalis and provide experimental method for quality standard of processed Morinda officinalis. METHODS: HPLC method was employed and Sinergi 4u fusion-RP (Phenomenex, 250 mm x 4. 6 mm, 4 microm) was used at column temperature of 25 degrees C. The mobile phase was composed of acetonitrile--0.4% phosphoric acid solution by gradient elution. The experiment data was analyzed by computer aided similarity evaluation software and statistic methods to draw total pattern of anthraquinone components of Morinda officinalis from different places. At the same time, under the same experimental condition, the qualities and quantities of different processed Morinda officinalis was analyzed and compared. And the total pattern of anthraquinones of different processed Morinda officinalis was drew. RESULTS: There were 20 common characteristic peaks which were demonstrated by using fingerprints similarity evaluating software both in Morinda officinalis from different places and different processed samples from Gaoliang, Guangdong. The data showed that anthraquinones of Morinda officinalis processed by different ways were similar but their relative contents were significantly different. And the 12th peaks was 1,8-dihydroxy anthraquinone. CONCLUSION: The contents of anthraquinones are influenced by different processed ways. The materials and temperature in the concocted procedure are the main influential factors. The fingerprints shows that the content of anthraquinones of Morinda officinalis processed by Glycyrrhiza uralensis Fisch. change much more obviously than that by alcohol and by salt. In addition, the fried processing method affects more obviously than the steamed one.
Assuntos
Antraquinonas/análise , Cromatografia Líquida de Alta Pressão/métodos , Medicamentos de Ervas Chinesas/química , Morinda/química , Tecnologia Farmacêutica/métodos , Antraquinonas/isolamento & purificação , Medicamentos de Ervas Chinesas/isolamento & purificação , Temperatura Alta , Raízes de Plantas/química , Plantas Medicinais/química , Controle de Qualidade , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To study the causes, clinical manifestations, treatment and prevention of calculus that develops in the prostatic cavity after transurethral resection of the prostate. METHODS: We reported 11 cases of calculus that developed in the prostatic cavity after transurethral resection or transurethral plasmakinetic resection of prostate. The patients complained of repeated symptoms of frequent micturition, urgent micturition and urodynia after operation, accompanied with urinary tract infection and some with urinary obstruction, which failed to respond to anti-infective therapies. Cystoscopy revealed calculi in the prostatic cavity, with eschar, sphacelus, uneven wound surface and small diverticula in some cases. After diagnosis, 1 case was treated by holmium laser lithotripsy and a second transurethral resection of the prostate, while the other 10 had the calculi removed under the cystoscope, followed by 1 -2 weeks of anti-infective therapy. RESULTS: After treatment, all the 11 cases showed normal results of routine urinalysis, and no more symptoms of frequent micturition, urgent micturition and urodynia. Three- to six-month follow-up found no bladder irritation symptoms and urinary tract infection. CONCLUSION: Repeated symptoms of frequent micturition, urgent micturition, urodynia and urinary tract infection after transurethral resection of the prostate should be considered as the indicators of calculus in the prostatic cavity, which can be confirmed by cystoscopy. It can be treated by lithotripsy or removal of the calculus under the cystoscope, or even a second transurethral resection of the prostate. For its prevention, excessive electric coagulation and uneven wound surface should be avoided and anti-infection treatment is needed.