RESUMO
OBJECTIVE: To evaluate the efficacy of Dongbai Tonglin Mixture (DTM) in the treatment of chronic prostatitis (CP) with the damp-heat downward diffusion syndrome. METHODS: We randomly selected 76 cases of CP with the damp-heat downward diffusion syndrome, equally divided them into a DTM and a control group, and treated them by oral administration of DTM and Qianlie Tai Tablets, respectively, both for 8 weeks. We obtained the NIH-CPSI and TCM Syndrome Scores of the patients, recorded the counts of white blood cells (WBC) and small particles of lecithin (SPL) in the prostate fluid, and compared them between the two groups before and after treatment. RESULTS: Compared with the baseline, the total NIH-CPSI scores were significantly reduced in both groups after treatment (P<0.05), particularly the scores on urination symptoms, pain / discomfort and quality of life (P<0.05), even more significantly in the DTM than in the control group (P<0.05), and so were the TCM Syndrome Scores (P<0.05), especially the scores on urinary incontinence, abdominal pain, perineal pain, and scrotal dampness (P<0.05), even more significantly in the former than in the latter group (P<0.05). The count of WBC in the prostate fluid was remarkably decreased (P<0.05), while that of SPL markedly increased in both groups after treatment (P<0.05), with an even more significant improvement in the DTM than in the control group (P<0.05), and the overall effectiveness rate of treatment was significantly higher in the former group than in the latter (88.89% vs 70.27%, P<0.05). CONCLUSION: Dongbai Tonglin Mixture is effective for the treatment of CP with the damp-heat downward diffusion syndrome.
Assuntos
Medicamentos de Ervas Chinesas , Fitoterapia , Prostatite , Masculino , Humanos , Prostatite/tratamento farmacológico , Medicamentos de Ervas Chinesas/uso terapêutico , Medicamentos de Ervas Chinesas/administração & dosagem , Adulto , Doença Crônica , Adulto Jovem , Resultado do Tratamento , Pessoa de Meia-Idade , Medicina Tradicional Chinesa/métodosRESUMO
A metal-free and efficient approach for the synthesis of structurally important nicotinates through 4-HO-TEMPO-mediated [3 + 3] annulation of cyclopropanols with ß-enamine esters is presented. This protocol features high atom efficiency, green waste, simple operation and broad substrate scope. Moreover, the experiments of gram-scale synthesis and recovery of oxidants make this strategy more sustainable and practical.
RESUMO
OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Ciliao" (BL 32) and "Huiyang" (BL 35) on the pain, urodynamic and the expressions of transient receptor poteintial vanilloid 1 (TRPV1) and P2X3 receptors in bladder of rats with interstitial bladder (IC), and to explore the possible mechanism on EA for IC. METHODS: A total of 24 Wistar female rats were randomly divided into a blank group, a model group and an EA group, 8 rats in each group. In the model group and the EA group, IC model was established by intraperitoneal injection of cyclophosphamide by 150 mg/kg at once. EA was applied at "Ciliao" (BL 32) and "Huiyang" (BL 35) in the EA group for 20 min, with continuous wave, 30 Hz in frequency, once a day for 3 consecutive days. Mechanical pain threshold of bladder and urodynamic indexes (first urination time, bladder effective volume and urination pressure) were observed after model establishment and after intervention, the expressions of TRPV1 and P2X3 receptors in the bladder were detected by Western blot. RESULTS: After model establishment, the mechanical pain threshold of bladder was decreased in the model group and the EA group compared with that in the blank group (P<0.01). After intervention, the mechanical pain threshold of bladder in the model group was lower than the blank group (P<0.01), and that in the EA group was higher than the model group (P<0.01). The urodynamic of the rats in the blank group was normal, obvious abnormal contraction during the filling period of bladder was found in the rats of the model group, while no abnormal contraction during the filling period was found in the rats of the EA group. After model establishment, in the model group and the EA group, the first urination time was earlier than the blank group (P<0.01), while bladder effective volume and urination pressure were lower than the blank group (P<0.01). After intervention, in the model group, the first urination time was earlier than the blank group (P<0.01), while bladder effective volume and urination pressure were lower than the blank group (P<0.05); in the EA group, the first urination time was later than the model group (P<0.05), while bladder effective volume and urination pressure were higher than the model group (P<0.05). Compared with the blank group, the protein expressions of TRPV1 and P2X3 receptors in bladder were up-regulated in the model group (P<0.01); compared with the model group, the protein expressions of TRPV1 and P2X3 receptors in bladder were down-regulated in the EA group (P<0.05). CONCLUSION: EA can relieve bladder pain and improve urodynamic in IC rats. The mechanism may be related to the down-regulation on the expressions of TRPV1 and P2X3 receptors and the further inhibition on the abnormal input of bladder signal.
Assuntos
Antineoplásicos , Cistite Intersticial , Eletroacupuntura , Ratos , Feminino , Animais , Cistite Intersticial/genética , Cistite Intersticial/terapia , Bexiga Urinária , Receptores Purinérgicos P2X3/genética , Receptores Purinérgicos P2X3/metabolismo , Ratos Sprague-Dawley , Ratos Wistar , Dor , Canais de Cátion TRPV/genética , Canais de Cátion TRPV/metabolismoRESUMO
Tumor necrosis factor-α-induced protein-8 like-2 (TIPE2) as a novel negative immune regulator plays an important role in several human diseases. However, its influences in cervical cancer and preeclampsia (PE) remain unclear. This study aims to explore the important role of TIPE2 in cervical cancer and PE via regulating cell invasion. TIPE2 expression in the cervical cancer tissues or the placenta of PE patients was detected. Human cervical cancer cell lines and trophoblasts were transfected with adenovirus expressing human TIPE2 and green fluorescent protein (GFP) (Ad-TIPE2), or the control adenovirus expressing GFP (Ad-GFP). Xenograft models were also constructed on nude mice, aiming to clarify how TIPE2 affects in vivo growth of cervical cancer cells. TIPE2 was down-regulated in the tumor tissues or placenta of patients with cervical cancer or PE. As a result, CaSKi and Hela cells in the Ad-TIPE2 group had decreased migration and invasion, with significant up-regulations of TIPE2 and E-cadherin, but down-regulations of ß-catenin and N-cadherin. Ad-TIPE2 decreased the volume and weight of xenograft tumors in the nude mice, with the down-regulation of Ki67. The quantity of cells (HTR8/SVneo and JEG3 cells) transfected with Ad-TIPE2 had increased, with up-regulations of TIPE2, matrix metalloproteinase (MMP)-2 and MMP-9. TIPE2 overexpression could reduce the invasion and migration of cervical cancer cells via inhibiting the epithelial-mesenchymal transition (EMT) process, and promote trophocyte invasion via upregulating the expression of MMPs, and it may be used as a potential therapeutic target for cervical cancer and PE.
Assuntos
Pré-Eclâmpsia , Neoplasias do Colo do Útero , Animais , Biomarcadores Tumorais , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células , Feminino , Células HeLa , Humanos , Peptídeos e Proteínas de Sinalização Intracelular , Camundongos , Camundongos Nus , Pré-Eclâmpsia/genética , Gravidez , Fator de Necrose Tumoral alfa , Neoplasias do Colo do Útero/genéticaRESUMO
A radical [1,4]-oxygen-atom transfer has been realized by the reaction of linear alkyne-tethered ketoximes and ethynylbenziodoxolones (EBX) under sequential catalytic conditions. Mechanism studies indicate that the O atom transfer experiences a cascade O atom radical cyclization/alkynylation/N-O bond photocleavage and subsequent N,O-diradical rearrangement. By the diversification of catalytic sequences, a series of structurally important 3H-pyrrol-3-ones and chlorinated furo[3,2-b]pyrroles are divergently synthesized along with an O atom shift under the catalysis of Cu/Ir photosensitization and Cu/Ir photosensitization/AlCl3, respectively.
RESUMO
Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a common disease in men, which has a high incidence and seriously affects the quality of life of the patients. The possible pathogenic factors of the disease include urine reflux, hidden infection, central sensitization, oxidative stress, and abnormal immune response, among which abnormal immune response plays a significant role in its development and progression. Immune response involves innate immunity and adaptive immunity, and most previous studies focused on adaptive immunity. In recent years, more and more attention has been paid to the role of the innate immune system in the pathogenesis of CP/CPPS. Studies show that mast cells, macrophages, Toll-like receptors and related cytokines in the innate immune system are all involved in the development and progression of CP/CPPS. As the innate immune system is the first barrier of the immune response of the body, studies on innate immunity will provide some new ideas for the diagnosis and treatment of CP/CPPS.
Assuntos
Dor Crônica , Prostatite , Masculino , Humanos , Prostatite/diagnóstico , Doença Crônica , Qualidade de Vida , Dor Pélvica/etiologia , Imunidade InataRESUMO
Surgical pleth index (SPI) has been widely investigated in assessing the nociceptive level. The aim of this study was to investigate the relationship between SPI level and patient responses to trachea intubation and skin incision. A total of 40 patients undergoing open abdominal general surgery were recruited for analyses. The patients were monitored with electrocardiogram, non-invasive blood pressure, SpO2, invasive blood pressure and SPI before anesthesia induction. Anesthesia was induced with midazolam, propofol, sufentanil and rocuronium and maintained with sufentanil and sevoflurane. Blood pressure, heart rate and SPI were recorded for analyses during the peri-intubation and peri-incision periods. A receiver operating characteristic (ROC) curve analysis was performed to analyze the predictive value of blood pressure, heart rate (HR) and SPI for hemodynamic responses for trachea intubation and skin incision. SPI had a similar changing trend to systolic blood pressure (SBP) and diastolic blood pressure (DBP). The SPI level was linearly correlated with SBP, DBP and HR. SPI increased significantly after intubation and incision in patients with positive but not negative responses to intubation and incision. The ROC analysis showed that only SBP level is predictive of intubation responses. These data suggested that SPI elevated under the noxious stimulation by intubation and incision, but it was not predictive of the hemodynamic responses to intubation and incision.
Assuntos
Hemodinâmica , Traqueia , Anestesia Geral , Pressão Sanguínea , Frequência Cardíaca , Humanos , Intubação IntratraquealRESUMO
A convenient and efficient vicinal oxyalkynylation/aminoalkynylation of internal unactivated alkenes is achieved by means of a Cu-catalyzed radical cascade reaction of unsaturated ketoximes with ethynylbenziodoxolone (EBX) reagents. This protocol enables the synthesis of structurally valuable isoxazolines or cyclic nitrones and the introduction of an important alkyne group in a single operation. The reaction is characterized by a broad substrate scope for both unsaturated ketoximes and alkynylation reagents and a low catalyst loading.
RESUMO
OBJECTIVE: To investigate the effect and safety of oral Tonglin Powder in the treatment of benign prostatic hyperplasia (BPH). METHODS: We conducted a randomized controlled study on 100 BPH patients, aged 40ï¼85 years, treated with Tonglin Powder (treatment group, n=50) or terazosin (control group, n=50), all for 3 months. Then we obtained the International Prostate Symptom Score (IPSS), quality of life score (QoL), prostate volume, postvoid residual urine volume (PVR), urine routine indexes, and liver and kidney function indexes from the patients and compared them between the two groups before and after treatment. RESULTS: The baseline data of the patients in the treatment and control groups were as follows, IPSS (22.24±7.33) vs (21.40±8.24), QoL 4 (2ï¼6) vs 4 (2ï¼6), prostate length 45 (30ï¼65) vs 45 (39ï¼65) mm, prostate width 35 (21ï¼54) vs 36 (26ï¼57) mm, and PVR 10 (5ï¼100) vs 10 (10ï¼100) ml, none with statistically significant difference between the two groups (P>0.05). After treatment, the patients of the treatment group, in comparison with those of the control, showed remarkable decreases in IPSS (11.60±6.49 vs 15.38±7.34, P=0.008) and QoL (2 ï¼»0ï¼5ï¼½ vs 3 ï¼»1ï¼6ï¼½, P=0.01). No statistically significant differences were observed between the treatment and control groups in prostate length (47 ï¼»38ï¼67ï¼½ vs 47.5 ï¼»38ï¼67ï¼½ mm), prostate width (36 ï¼»26ï¼57ï¼½ vs 36.5 ï¼»31ï¼57ï¼½ mm), and PVR (10 ï¼»8ï¼100ï¼½ vs 10 ï¼»8ï¼70ï¼½ ml) (P>0.05). The Nimodipine method of evaluation showed that the excellence rate of therapeutic effectiveness was significantly higher in the treatment than in the control group (40% vs 8%, P<0.001), and so was the total effectiveness rate (82% vs 64%, P=0.043). CONCLUSIONS: Tonglin Powder can effectively improve the symptoms of BPH, such as difficult urination, and hence the patient's quality of life.
Assuntos
Medicamentos de Ervas Chinesas/uso terapêutico , Hiperplasia Prostática/tratamento farmacológico , Adulto , Idoso , Idoso de 80 Anos ou mais , Humanos , Masculino , Pessoa de Meia-Idade , Prazosina/análogos & derivados , Prazosina/uso terapêutico , Qualidade de Vida , Resultado do TratamentoRESUMO
OBJECTIVE: To study effects of blue light irradiation on monosaccharide composition of intracellular polysacchride and contents of cordycepin and cordyceps polysacchride of mycelium and sporocarp in Cordyceps militaris. METHODS: The monosaccharide composition of intracellular polysacchride of mycelium and sporocarp in Cordyceps militaris as materials were determined by gas chromatography after 144 h blue light irradiation. The contents of cordycepin and cordyceps polysacchride of mycelium and sporocarp in Cordyceps militaris were detected at different blue light irradiation periods. At the same time, the growth of mycelium and sporocarp in Cordyceps militaris were observed during blue light irradiation. RESULTS: Mycelium polysaccharide in Cordyceps militaris was a kind of heteropolysaccharide containing four kinds of monosaccharide and fruiting body polysaccharide was a kind of heteropolysaccharide containing five kinds of monosaccharide. Whether blue light irradiation or dark culture, the content changes of cordyceps polysacchride in two groups showed similar patterns in the test of mycelium polysaccharides. The content changes of cordyceps polysacchride in two groups were basically the same in the detection of sporocarp polysacchride. Cordycepin content in the two set of experiments of blue light irradiation all showed a clear upward trend in the detection of mycelium and sporocarp in Cordyceps militaris. CONCLUSION: The blue light irradiation has certain effect on the species and quantity of monosaccharide in intracellular polysaccharide. The content increase of cordycepin and cordyceps polysacchride in Cordyceps militaris are promoted by blue light irradiation. Blue light can help the morphogenesis and promote the differentiation and growth of sporocarp in Cordyceps militaris. This study is the first report about the effect of blue light on the type and quantity of the monosaccharide composition in polysaccharide of Cordyceps militaris, which will lay the foundation for further study on the metabolism of active substance in Cordyceps militaris by blue light irradiation.
Assuntos
Cordyceps/química , Desoxiadenosinas/química , Monossacarídeos/química , Polissacarídeos/química , Espaço Extracelular/química , Luz , MicélioRESUMO
Agarase hydrolyzes agarose into a series of oligosaccharides with repeating disaccharide units. The glycoside hydrolase (GH) module of agarase is known to be responsible for its catalytic activity. However, variations in the composition of the GH module and its effects on enzymatic functions have been minimally elucidated. The agaG4 gene, cloned from the genome of the agarolytic Flammeovirga strain MY04, encodes a 503-amino acid protein, AgaG4. Compared with elucidated agarases, AgaG4 contains an extra peptide (Asn(246)-Gly(302)) within its GH module. Heterologously expressed AgaG4 (recombinant AgaG4; rAgaG4) was determined to be an endo-type ß-agarase. The protein degraded agarose into neoagarotetraose and neoagarohexaose at a final molar ratio of 1.5:1. Neoagarooctaose was the smallest substrate for rAgaG4, whereas neoagarotetraose was the minimal degradation product. Removing the extra fragment from the GH module led to the inability of the mutant (rAgaG4-T57) to degrade neoagarooctaose, and the final degradation products of agarose by the truncated protein were neoagarotetraose, neoagarohexaose, and neoagarooctaose at a final molar ratio of 2.7:2.8:1. The optimal temperature for agarose degradation also decreased to 40 °C for this mutant. Bioinformatic analysis suggested that tyrosine 276 within the extra fragment was a candidate active site residue for the enzymatic activity. Site-swapping experiments of Tyr(276) to 19 various other amino acids demonstrated that the characteristics of this residue were crucial for the AgaG4 degradation of agarose and the cleavage pattern of substrate.
Assuntos
Glicosídeo Hidrolases/química , Peptídeos/química , Sefarose/química , Sequência de Aminoácidos , Bactérias/genética , Catálise , Domínio Catalítico , Clonagem Molecular , Biologia Computacional/métodos , Primers do DNA/genética , Vetores Genéticos , Modelos Genéticos , Dados de Sequência Molecular , Mutação , Oligossacarídeos/química , Filogenia , Ligação Proteica , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , TemperaturaAssuntos
Bandagens , Queimaduras/tratamento farmacológico , Óxido Nitroso/administração & dosagem , Oxigênio/administração & dosagem , Medição da Dor/efeitos dos fármacos , Administração por Inalação , Adolescente , Adulto , Idoso , Bandagens/efeitos adversos , Queimaduras/patologia , Método Duplo-Cego , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Medição da Dor/métodos , Resultado do Tratamento , Adulto JovemRESUMO
BACKGROUND: Pain is a major problem for patients with advanced cancer and one of the most frequent and disturbing of all cancer-related symptoms. Researchers continue to report that cancer pain remains undertreated. Inadequate pain control can significantly affect the patient's quality of life and may in turn affect the patient's will to live or comply with treatment recommendations. A better understanding of the experience of cancer pain management is important in identifying factors responsible for undertreated pain. OBJECTIVE: This study aimed to obtain the experience of cancer pain management. INTERVENTIONS: We used a phenomenological approach to explore the status of cancer pain management through participants' experience. Semistructured interviews were conducted with 14 family caregivers, patients, and acquaintances and 14 health professionals (nurses and physicians) from a regional tertiary hospital in northwest China. Data were collected by in-depth interviews. We used a qualitative description after full transcription of every interview. Analysis involved the identification of themes and the development of a taxonomy of participants' experience of cancer pain management. RESULTS: Taxonomy used in this study is to identify, code, group, and name meaning units of the transcribed interviews by reading through repeatedly to obtain an initial sense. Four themes were identified: (1) marginalization, (2) hopelessness and helplessness, (3) deficiency of access and resources, and (4) expectations related to pain. CONCLUSION: Findings from this study suggest that the situation of patients with undertreated cancer pain continues. IMPLICATIONS FOR PRACTICE: Special attention should be paid by policymakers, professionals, and family caregivers to the marginalized group of cancer patients who suffer with pain.
Assuntos
Área Carente de Assistência Médica , Neoplasias/complicações , Manejo da Dor/métodos , Dor Intratável/etiologia , Dor Intratável/terapia , Adulto , Idoso , Cuidadores/estatística & dados numéricos , China , Estudos Transversais , Feminino , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Serviços de Assistência Domiciliar/normas , Serviços de Assistência Domiciliar/tendências , Humanos , Entrevistas como Assunto , Masculino , Pessoa de Meia-Idade , Avaliação das Necessidades , Neoplasias/diagnóstico , Neoplasias/terapia , Manejo da Dor/tendências , Dor Intratável/fisiopatologia , Serviços de Saúde Rural/normas , Serviços de Saúde Rural/tendências , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
Granular Mg/Al mixed oxides (G-Mg3.3 AlO4.8) was successfully synthesized by using Mg/Al layered double hydroxide (Mg/Al-LDH) as the precursor. Its structure was characterized by analytical methods of BET, XRD, SEM and FT-IR. Adsorption properties of thiocyanate anion (SCN-) on this new material were also investigated. Specific surface area and average pore width of G-Mg3.3 AlO4.8 are 269.4 m2 x g(-1) and 13.25 nm respectively. Molding and granulation retains the layer structure and generates no new phase. Freundlich isotherm equation and pseudo-second-order kinetic equation can describe the static adsorption process precisely and adsorption capacity reaches 165.8 mg x g(-1) at 25 degrees C and 500 mg x L(-1). Yoon-Nelson model can predict the breakthrough curves of SCN- on G-Mg3.3 AlO4.8 precisely. The breakthrough adsorption capacity reaches 50.73 mg x g(-1) under the conditions with initial concentration is 100 mg x L(-1), flow rate is 5 mL x min(-1), bed layer altitude is 10 cm and initial pH is 6. The removal rates of SCN- are all above 98% after reusing four times. Consequently, G-Mg3.3 AlO4. 8 is a reusable and high-efficient adsorbent.
Assuntos
Compostos de Alumínio/química , Compostos de Magnésio/química , Tiocianatos/isolamento & purificação , Poluentes Químicos da Água/isolamento & purificação , Purificação da Água/métodos , Adsorção , Ânions/isolamento & purificaçãoRESUMO
Enhanced ganoderic acid Me (GA-Me, an important anti-tumor triterpene) yield was attained with the medicinal fungus Ganoderma lucidum using response surface methodology (RSM). Interactions were studied with three variables, viz. glucose, peptone and culture time using a Central Composite Design (CCD). The CCD contains a total of 20 experiments with the first 14 experiments organized in a fractional factorial design, with the experimental trails from 15 to 20 involving the replications of the central points. A polynomial model, describing the relationships between the yield of GA-Me and the three factors in a second-order equation, was developed. The model predicted the maximum GA-Me yield of 11.9 mg·L−1 for glucose, peptone, culture time values of 44.4 g·L−1, 5.0 g·L−1, 437.1 h, respectively, and a maximum GA-Me yield of 12.4 mg·L−1 was obtained in the validation experiment, which represented a 129.6% increase in titre compared to that of the non-optimized conditions. In addition, 11.4 mg·L−1 of GA-Me was obtained in a 30-L agitated fermenter under the optimized conditions, suggesting the submerged culture conditions optimized in the present study were also suitable for GA-Me production on a large scale.
Assuntos
Fermentação , Reishi/metabolismo , Triterpenos/metabolismo , Algoritmos , Análise de Variância , Reatores Biológicos , Meios de Cultura/química , Técnicas de Cultura , Medicamentos de Ervas Chinesas/metabolismo , Modelos Biológicos , Análise de Regressão , Reishi/crescimento & desenvolvimentoAssuntos
Leiomiossarcoma , Omento , Neoplasias Peritoneais , Adulto , Doenças Assintomáticas , Humanos , Achados Incidentais , Laparoscopia , Laparotomia , Leiomiossarcoma/diagnóstico , Leiomiossarcoma/patologia , Leiomiossarcoma/cirurgia , Masculino , Omento/patologia , Omento/cirurgia , Neoplasias Peritoneais/diagnóstico , Neoplasias Peritoneais/patologia , Neoplasias Peritoneais/cirurgia , UltrassonografiaRESUMO
A metagenomic library was prepared using pCC2FOS vector containing about 3.0 Gbp of community DNA from the microbial assemblage of activated sludge. Screening of a part of the un-amplified library resulted in the finding of 1 unique lipolytic clone capable of hydrolyzing tributyrin, in which an esterase gene was identified. This esterase/lipase gene consists of 834 bp and encodes a polypeptide (designated EstAS) of 277 amino acid residuals with a molecular mass of 31 kDa. Sequence analysis indicated that it showed 33% and 31% amino acid identity to esterase/lipase from Gemmata obscuriglobus UQM 2246 (ZP_02733109) and Yarrowia lipolytica CLIB122 (XP_504639), respectively; and several conserved regions were identified, including the putative active site, HSMGG, a catalytic triad (Ser92, His125 and Asp216) and a LHYFRG conserved motif. The EstAS was overexpressed, purified and shown to hydrolyse p-nitrophenyl (NP) esters of fatty acids with short chain lengths (< or = C8). This EstAS had optimal temperature and pH at 35 degrees C and 9.0, respectively, by hydrolysis of p-NP hexanoate. It also exhibited the same level of stability over wide temperature and pH ranges and in the presence of metal ions or detergents. The high level of stability of esterase EstAS with its unique substrate specificities make itself highly useful for biotechnological applications.
Assuntos
Esterases/genética , Esgotos/microbiologia , Sequência de Aminoácidos , Domínio Catalítico , Clonagem Molecular , DNA Bacteriano/metabolismo , Esterases/metabolismo , Concentração de Íons de Hidrogênio , Metagenoma , Dados de Sequência Molecular , Alinhamento de Sequência , Homologia de Sequência de Aminoácidos , Especificidade por Substrato , Temperatura , Triglicerídeos/metabolismo , Yarrowia/classificação , Yarrowia/enzimologiaRESUMO
Protein phosphatases-2A (PP-2A) is a major serine/threonine phosphatase and accounts for more than 50% serine/threonine phosphatase activity in eukaryotes. The holoenzyme of PP-2A consists of the scaffold A subunit, the catalytic C subunit and the regulatory B subunit. The scaffold subunits, PP2A-A alpha/beta, provide a platform for both C and B subunits to bind, thus playing a crucial role in providing specific PP-2A activity. Mutation of the two genes encoding PP2A-A alpha/beta leads to carcinogenesis and likely other human diseases. Regulation of these genes by various factors, both extracellular and intracellular, remains largely unknown. In the present study, we have conducted functional dissection of the promoter of the mouse PP2A-A alpha gene. Our results demonstrate that the proximal promoter of the mouse PP2A-A alpha gene contains numerous cis-elements for the binding of CREB, ETS-1, AP-2 alpha, SP-1 besides the putative TFIIB binding site (BRE) and the downstream promoter element (DPE). Gel mobility shifting assays revealed that CREB, ETS-1, AP-2 alpha, and SP-1 all bind to PP2A-A alpha gene promoter. In vitro mutagenesis and reporter gene activity assays reveal that while SP-1 displays negative regulation, CREB, ETS-1 and AP-2A alpha all positively regulate the promoter of the PP2A-A alpha gene. ChIP assays further confirm that all the above transcription factors participate the regulation of PP2A-A alpha gene promoter. Together, our results reveal that multiple transcription factors regulate the PP2A-A alpha gene.
Assuntos
Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Regulação Enzimológica da Expressão Gênica , Proteína Fosfatase 2/metabolismo , Proteína Proto-Oncogênica c-ets-1/metabolismo , Fator de Transcrição Sp1/metabolismo , Fator de Transcrição AP-2/metabolismo , Animais , Sequência de Bases , Sítios de Ligação , Domínio Catalítico , Humanos , Camundongos , Dados de Sequência Molecular , Regiões Promotoras Genéticas , Fatores de Transcrição/metabolismoRESUMO
Extracellular alginate lyase secreted by Vibrio sp. QY101, which was isolated from brown algae, was purified to homogeneity by a combination of ammonium sulfate precipitation, DEAE-Sepharose Fast Flow anion-exchange chromatography and Superdex 75 gel filtration chromatography. Its molecular mass was 39 kD as determined by SDS-PAGE analysis. The enzyme had an optimal temperature of 30 degrees for its activity, and was most active at pH 7.5. The thermal and pH stability were 0-30 degrees, and pH 6.5-8.5, respectively. The enzyme activity was stimulated by 0.5 mol/L NaCl, 1.0 mmol/L Ca(2+) or 5.0 mmol/L (Mn(2+), and inhibited by 5.0 mmol/L Ni(2+), 1.0 mmol/L Fe2+) or 1.0 mmol/L EDTA. Preliminary analysis on substrate specificity showed that this alginate lyase had activity on both poly-alpha 1,4-L-guluronate and poly-beta1,4-D-mannuronate substrates.