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BACKGROUNDS & AIMS: Portal hypertension (PH) is one of the most frequent complications of chronic liver disease. The peripheral 5-hydroxytryptamine (5-HT) level was increased in cirrhotic patients. We aimed to elucidate the function and mechanism of 5-HT receptor 1A (HTR1A) in the portal vein (PV) on PH. METHODS: PH models were induced by thioacetamide injection, bile duct ligation, or partial PV ligation. HTR1A expression was detected using real-time polymerase chain reaction, in situ hybridization, and immunofluorescence staining. In situ intraportal infusion was used to assess the effects of 5-HT, the HTR1A agonist 8-OH-DPAT, and the HTR1A antagonist WAY-100635 on portal pressure (PP). Htr1a-knockout (Htr1a-/-) rats and vascular smooth muscle cell (VSMC)-specific Htr1a-knockout (Htr1aΔVSMC) mice were used to confirm the regulatory role of HTR1A on PP. RESULTS: HTR1A expression was significantly increased in the hypertensive PV of PH model rats and cirrhotic patients. Additionally, 8-OH-DPAT increased, but WAY-100635 decreased, the PP in rats without affecting liver fibrosis and systemic hemodynamics. Furthermore, 5-HT or 8-OH-DPAT directly induced the contraction of isolated PVs. Genetic deletion of Htr1a in rats and VSMC-specific Htr1a knockout in mice prevented the development of PH. Moreover, 5-HT triggered adenosine 3',5'-cyclic monophosphate pathway-mediated PV smooth muscle cell contraction via HTR1A in the PV. We also confirmed alverine as an HTR1A antagonist and demonstrated its capacity to decrease PP in rats with thioacetamide-, bile duct ligation-, and partial PV ligation-induced PH. CONCLUSIONS: Our findings reveal that 5-HT promotes PH by inducing the contraction of the PV and identify HTR1A as a promising therapeutic target for attenuating PH. As an HTR1A antagonist, alverine is expected to become a candidate for clinical PH treatment.
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BACKGROUND: The triglyceride-glucose (TyG) index serves as a reliable proxy for insulin resistance (IR). IR has been linked to heightened incidence, prevalence, or severity of chronic obstructive pulmonary disease (COPD) and asthma. Prior research indicates that critically ill patients are prone to developing IR. Nevertheless, few studies have delved into the correlation between IR and all-cause mortality in critically ill patients with COPD and asthma. Therefore, the aim of this study is to explore the association between the TyG index and all-cause mortality in patients with COPD and asthma, with the goal of assessing the impact of IR on the prognosis of this patient population. METHODS: This is a retrospective study, and all data are from the Medical Information Mart for Intensive Care IV (MIMIC-IV) critical care database. This study included 684 ICU patients with COPD and asthma and divided them into quartiles based on TyG index levels. The primary outcomes of this study were all-cause mortality during follow-up, encompassing mortality at 30 days, 90 days, and 180 days. The Kaplan-Meier analysis was used to compare all-cause mortality among the above four groups. Cox proportional hazards analyses were performed to examine the association between TyG index and all-cause mortality in critically ill patients with COPD and asthma. Restricted cubic spline analysis was used to assess potential nonlinear association between the TyG index and the primary outcome. RESULTS: A total of 684 patients (53.9% female) were included. The 90-days all-cause mortality rate and 180-days all-cause mortality were 11.7% and 12.3%, respectively. Kaplan-Meier analysis revealed a significant association between the TyG index and both 90-days all-cause mortality (log-rank p = .039) and 180-days all-cause mortality (log-rank p = .017). Cox proportional hazards analysis revealed a significant association between the TyG index and 90-days all-cause mortality in both the unadjusted model (HR, 1.30 [95% CI 1.08-1.57] p = .005) and the model adjusted for age, gender, and diabetes (HR, 1.38 [95% CI 1.15-1.67] p < .001). Similarly, the TyG index was associated with 180-days all-cause mortality in the unadjusted model (HR, 1.30 [95% CI 1.09-1.56] p = .004) and the model adjusted for age, sex, and diabetes (HR, 1.38 [95% CI 1.15-1.66] p < .001). The restricted cubic splines (RCS) regression model indicated a significant nonlinear association between the TyG index and both 90-days and 180-days all-cause mortality. Specifically, TyG index >4.8 was associated with an increased risk of mortality at both 90 days and 180 days. CONCLUSIONS: In summary, our results extend the utility of the TyG index to critically ill patients with COPD and asthma. Our study shows that the TyG index is a potential predictor of all-cause mortality in critically ill patients with COPD and asthma. In addition, in patients with a TyG index exceeding 4.8, there was a heightened risk of mortality. Measuring the TyG index may help with risk stratification and prognosis prediction in critically ill patients with COPD and asthma. Further prospective studies are needed to confirm our findings.
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Asma , Diabetes Mellitus , Enfermedad Pulmonar Obstructiva Crónica , Humanos , Femenino , Masculino , Estudios Retrospectivos , Enfermedad Crítica , GlucosaRESUMEN
BACKGROUND: This study aimed at developing and validating a scoring model to stratify critically ill patients after cardiac surgery based on risk for dysphagia, a common but often neglected complication. METHODS: Data were prospectively collected and analyzed from January 2016 to June 2017 from 395 consecutive post cardiac surgery patients at the cardiac care unit (CCU) at a single center; 103 (26.1%) developed dysphagia. Univariate and multivariate logistic analyses were used to identify independent predictors for dysphagia. The survival nomogram was developed on the basis of a multivariable Cox model, which allowed us to obtain survival probability estimations. The predictive performance of the nomogram was verified for discrimination and calibration. Areas under receiver operating characteristic curve analysis were used to illustrate and evaluate the diagnostic performance of the novel model. RESULTS: The final novel scoring model, named SSG-OD, consists of three independent factors: gastric intubation (OR = 1.024, 95% CI 1.015-1.033), sedative drug use duration (OR = 1.031, 95% CI 1.001-1.063) and stroke or not (OR = 6.182, 95% CI 3.028-12.617). SSG-OD identified patients at risk for dysphagia with sensitivity of 68.5% and specificity of 89.0% (OR = 0.833, 95% CI: 0.782-0.884). The positive and negative likelihood ratios were 6.22 and 0.35. CONCLUSIONS: The novel SSG-OD scoring system to risk stratify CCU patients for dysphagia is an easy-to-use bedside prognostication aid with good predictive performance and the potential to reduce aspiration incidence and accelerate recovery.
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Procedimientos Quirúrgicos Cardíacos/métodos , Enfermedad Crítica , Trastornos de Deglución/epidemiología , Complicaciones Posoperatorias/epidemiología , Anciano , Estudios de Cohortes , Trastornos de Deglución/etiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Nomogramas , Valor Predictivo de las Pruebas , Estudios Prospectivos , Sensibilidad y EspecificidadRESUMEN
PURPOSE: To determine if dynamic contrast-enhanced MRI (DCE-MRI) could correlate well with invasive angiography in the characterization of spinal tumor vascularity. METHODS: Totally 40 patients with untreated spinal tumors underwent MRI before preoperative angiography and embolization. Tumors were assigned to hypervascular, moderate, or hypovascular groups based on angiographic appearance. Tumor vascularity was also evaluated with enhancement degree on standard MR and with DCE-MRI parameters via ROI analysis of enhanced tumor area. The Spearman correlation coefficient was calculated to determine the correlation between the degree of angiographic vascularity and enhancement on MRI and DCE-MRI parameters. ROC analysis was conducted to assess the appropriate cut-off value. RESULTS: There were 12 hypervascular, 12 moderate, and 16 hypovascular tumors, respectively. The Spearman correlation coefficient between DCE-MRI parameter and the degree of angiographic vascularity was 0.899 (RSlopemax), 0.847 (Slopemax), 0.697 (E max), 0.694 (ERmax), and -0.587 (TTP), respectively, which showed excellent-to-moderate relationships. The RSlopemax cut-off value of 1.325 provided the highest specificity of 100 % and sensitivity of 87.5 % in predicting hypovascular tumors and the value of 1.85 provided the highest sensitivity of 100 % and specificity of 96.4 % in characterizing hypervascular ones. CONCLUSIONS: DCE-MRI is an accurate technique for the assessment of spinal tumor vascularity, which may have a potential value in the decision-making of preoperative embolization.
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Angiografía/métodos , Imagen por Resonancia Magnética/métodos , Neoplasias de la Médula Espinal/diagnóstico por imagen , Adolescente , Adulto , Anciano , Medios de Contraste , Femenino , Humanos , Masculino , Persona de Mediana Edad , Neovascularización Patológica/diagnóstico por imagen , Sensibilidad y Especificidad , Adulto JovenRESUMEN
The ß-globin matrix attachment regions (MARs) were inserted into the 5'-site of the eukaryotic expression vector cassette and DNA fragments 350 and 750 bp in length were inserted into the site to generate expression vectors with varying distances between the expression cassette and MAR. The vectors containing MARs increased chloramphenicol acetyltransferase (CAT) expression levels compared to the negative control vector lacking the MAR; the highest expression increase was 3.8-fold. A greater MAR-transgene distance (750 bp) correlated with a greater increase in transgene expression when compared to the control vector that lacked separation between the MAR and transgene. CAT gene copy numbers were higher in cells transformed with the vector possessing a smaller MAR-transgene distance (350 bp) than in cells belonging to the other three groups. However, MAR-induced transgene expression levels did not exhibit a direct relationship with gene copy number.
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Cloranfenicol O-Acetiltransferasa/genética , Cloranfenicol O-Acetiltransferasa/metabolismo , Expresión Génica , Regiones de Fijación a la Matriz , Transgenes , Globinas beta/genética , Animales , Células CHO , Cricetinae , Cricetulus , Dosificación de Gen , Regulación de la Expresión Génica , Vectores Genéticos/genética , Vectores Genéticos/metabolismo , Globinas beta/metabolismoRESUMEN
Scutellarin (SG) and its aglycone, Scutellarein (S), are flavonoids of therapeutic cardiocerebrovascular disease. SG was hydrolyzed by bacterial enzyme into S which was absorbed in the intestine. The aim of this study was to determine the effects of the microflora in the intestinal lumen and the efflux transporter of intestinal epithelial cells on the absorption process of SG and S. After oral administration of antibiotics in Sprague-Dawley rats, the reduced bacterial enzyme formation significantly hinders the absorption of SG, whereas scarcely that of S. The absorption study in situ single-pass intestinal perfusion revealed that S could be absorbed throughout the intestine of rats. The effective intestinal permeability of S in the jejunum was much lower than in the other sections of the GI tract. The efflux transporter promoted SG secretion into lumen from enterocytes, which hindered the absorption of both SG and S into the bloodstream. The efflux transporter protein inhibitor (verapamil, probenecid and reserpine) remarkably enhanced the absorption of S and the bioconversion of S into SG in both the rat intestine and Caco-2-monolayer models.
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Apigenina/farmacocinética , Mucosa Intestinal/metabolismo , Intestinos/microbiología , Microbiota , Administración Oral , Animales , Células CACO-2 , Glucuronatos , Humanos , Absorción Intestinal/efectos de los fármacos , Masculino , Proteínas de Transporte de Membrana/metabolismo , Ratas Sprague-DawleyRESUMEN
BACKGROUND: The low-dose computed tomography (CT) technique has been widely used because it decreases the potential risk of radiation exposure, as well as enabling low-dose CT-guided lung lesion biopsy. However, uncertainties remain regarding diagnostic accuracy, radiation dose, complication rate, and image quality. PURPOSE: To compare the diagnostic accuracy, radiation dose, complication rate, and image quality of lung lesion biopsy between conventional CT-guided and low-dose CT-guided techniques. MATERIAL AND METHODS: A total of 90 patients were prospectively enrolled and randomized into two groups (group A: 120 kv; 200 mA; thickness, 2.0 mm; pitch, 16 mm/rot; n = 44; group B: 120 kv;10 mA; thickness, 2.0 mm; pitch, 23 mm/rot; n = 46). Sensitivity, specificity, accuracy, positive predictive value (PPV), negative predictive value (NPV), radiation dose, image quality, and complication rate were compared. All variables between the two groups were analyzed using chi-square and Student's t tests. A P value of < 0.05 was considered statistically significant. RESULTS: The sensitivity, specificity, accuracy, positive predictive value (PPV), and negative predictive value (NPV) for diagnosing lung lesions were 96.88%, 100%, 97.5%, 100%, and 88.89% in group A, respectively. In group B, the values were 96.67%, 100%, 97.5%, 100%, and 90.91%, respectively (P > 0.05). The mean weighted CT dose index (CTDIw) and dose-length product (DLP) were 29.29 ± 3.93 mGy and 211.74 ± 37.89 mGy*cm in group A and 1.55 ± 0.15 mGy and 10.98 ± 1.56 mGy*cm in group B (P < 0.001). Image quality satisfied the need for a coaxial biopsy. Complications in group A and group B were observed in 27.28% and 23.91% of the patients, respectively (P > 0.05). CONCLUSION: Compared to conventional CT-guided biopsies, lung lesion biopsies guided by the low-dose CT biopsy protocol showed dramatically lower CTDIw and DLP levels. In contrast, the diagnostic yield of the procedures did not differ significantly, which is a recommended technique in certain populations.
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Neoplasias Pulmonares/diagnóstico por imagen , Neoplasias Pulmonares/patología , Dosis de Radiación , Radiografía Intervencional/métodos , Tomografía Computarizada por Rayos X/métodos , Adulto , Anciano , Anciano de 80 o más Años , Biopsia , Estudios de Factibilidad , Femenino , Humanos , Pulmón/diagnóstico por imagen , Pulmón/patología , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Estudios Prospectivos , Reproducibilidad de los Resultados , Sensibilidad y Especificidad , Adulto JovenRESUMEN
Objectives: The aim of this study is to assess the influence of cardiopulmonary coupling (CPC) based on RCMSE on the prediction of complications and death in patients with acute type A aortic dissection (ATAAD). Background: The cardiopulmonary system may be nonlinearly regulated, and its coupling relationship with postoperative risk stratification in ATAAD patients has not been studied. Methods: This study was a single-center, prospective cohort study (ChiCTR1800018319). We enrolled 39 patients with ATAAD. The outcomes were in-hospital complications and all-cause readmission or death at 2 years. Results: Of the 39 participants, 16 (41.0%) developed complications in the hospital, and 15 (38.5%) died or were readmitted to the hospital during the two-year follow-up. When CPC-RCMSE was used to predict in-hospital complications in ATAAD patients, the AUC was 0.853 (p < 0.001). When CPC-RCMSE was used to predict all-cause readmission or death at 2 years, the AUC was 0.731 (p < 0.05). After adjusting for age, sex, ventilator support (days), and special care time (days), CPC-RCMSE remained an independent predictor of in-hospital complications in patients with ATAAD [adjusted OR: 0.8 (95% CI, 0.68-0.94)]. Conclusion: CPC-RCMSE was an independent predictor of in-hospital complications and all-cause readmission or death in patients with ATAAD.
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BACKGROUND: For critically ill patients, physicians tend to administer sufficient or even excessive oxygen to maintain oxygen saturation at a high level. However, the credibility of the evidence for this practice is unclear. OBJECTIVE: To determine the effects of different oxygen therapy strategies on the outcomes of mechanically ventilated intensive care unit (ICU) patients. DESIGN AND SETTING: Systematic review of the literature and meta-analysis conducted at Jiangxi Provincial People's Hospital, Affiliated to Nanchang University, Nanchang, China. METHODS: We systematically searched electronic databases such as PubMed and Embase for relevant articles and performed meta-analyses on the effects of different oxygen therapy strategies on the outcomes of mechanically ventilated ICU patients. RESULTS: A total of 1802 patients from five studies were included. There were equal numbers of patients in the conservative and liberal groups (n = 910 in each group). There was no significant difference between the conservative and liberal groups with regard to 28-day mortality (risk ratio, RR = 0.88; 95% confidence interval, CI = 0.59-1.32; P = 0.55; I2 = 63%). Ninety-day mortality, infection rates, ICU length of stay, mechanical ventilation-free days up to day 28 and vasopressor-free days up to day 28 were comparable between the two strategies. CONCLUSIONS: It is not necessary to use liberal oxygen therapy strategies to pursue a higher level of peripheral oxygen saturation for mechanically ventilated ICU patients. Conservative oxygen therapy was not associated with any statistically significant reduction in mortality.
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Oxígeno , Respiración Artificial , Enfermedad Crítica/terapia , Humanos , Unidades de Cuidados Intensivos , Tiempo de Internación , Terapia por Inhalación de Oxígeno , PronósticoRESUMEN
BACKGROUND: Intravenous fluids are used commonly for almost all intensive care unit (ICU) patients, especially for patients in need of resuscitation. The selection and use of resuscitation fluids may affect the outcomes of patients; however, the optimal resuscitative fluid remains controversial. METHODS: We systematically searched PubMed, Embase, and CENTRAL. Studies comparing balanced crystalloids and normal saline in ICU patients were selected. We used the Cochrane Collaboration tool to assess the risk of bias in studies. The primary outcome was mortality at the longest follow-up. Secondary outcomes included the incidence of acute kidney injury (AKI) and new renal replacement therapy (RRT). RESULTS: A total of 35,456 patients from eight studies were included. There was no significant difference between balanced crystalloid solutions and saline in mortality (risk ratio [RR]: 0.96; 95% confidence interval [CI]:0.92-1.01). The subgroup analysis with traumatic brain injury (TBI) showed lower mortality in patients receiving normal saline (RR:1.25; 95% CI 1.02-1.54). However, in patients with non-TBI, balanced crystalloid solutions achieved lower mortality than normal saline (RR: 0.94; 95% CI 0.90-0.99). There was no significant difference in moderate to severe AKI (RR: 0.96; 95% CI 0.90-1.01) or new RRT (RR: 0.94; 95% CI 0.84-1.04). CONCLUSIONS: Compared with normal saline, balanced crystalloids may not improve the outcomes of mortality, the incidence of AKI, and the use of RRT for critically ill patients. However, balanced crystalloids reduce the risk of death in patients with non-TBI but increase the risk of death in those with TBI. Large-scale rigorous randomized trials with better designs are needed, especially for specific patient populations.
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Lesión Renal Aguda , Solución Salina , Lesión Renal Aguda/etiología , Lesión Renal Aguda/terapia , Enfermedad Crítica/terapia , Soluciones Cristaloides , Femenino , Fluidoterapia , Humanos , Soluciones Isotónicas/uso terapéutico , Masculino , Solución Salina/uso terapéuticoRESUMEN
Recombinant protein production by mammalian cells is the initial step in the manufacture of many therapeutic proteins. Chinese hamster ovary (CHO) cells are the most common host system to produce recombinant therapeutic proteins (RTPs). However, it is still challenging to maintain high productivity ensuring the good quality of RTPs produced by CHO cells. MicroRNAs(miRNAs) are short regulatory non-coding RNAs that can regulate cellular behavior and complex phenotypes. It has been found that miRNAs can enhance the expression level of recombinant proteins in CHO cells by promoting proliferation, resisting apoptosis, and regulating metabolism. miRNAs also can affect the quality of RTPs. In this review, we will discuss the effect and mechanism of miRNA on the expression level and quality of recombinant proteins in CHO cells.
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AIM: To study the metabolic and pharmacokinetic profile of scutellarin, an active component from the medical plant Erigeron breviscapus (Vant) Hand-Mazz, and to investigate the mechanisms underlying the low bioavailability of scutellarin though oral or intravenous administration in rats. METHODS: HPLC method was developed for simultaneous detection of scutellarin and scutellarein (the aglycone of scutellarin) in rat plasma, urine and feces. The in vitro metabolic stability study was carried out in rat liver microsomes from different genders. RESULTS: After a single oral dose of scutellarin (400 mg/kg), the plasma concentrations of scutellarin and scutellarein in female rats were significantly higher than in male ones. Between the female and male rats, significant differences in AUC, t(max2) and C(max2) for scutellarin were found. The pharmacokinetic parameters of scutellarin in the urine also showed significant gender differences. After a single oral dose of scutellarin (400 mg/kg), the total percentage excretion of scutellarein in male and female rats was 16.5% and 8.61%, respectively. The total percentage excretion of scutellarin and scutellarein in the feces was higher with oral administration than with intravenous administration. The in vitro t(1/2) and CL(int) value for scutellarin in male rats was significantly higher than that in female rats. CONCLUSION: The results suggest that a large amount of ingested scutellarin was metabolized into scutellarein in the gastrointestinal tract and then excreted with the feces, leading to the extremely low oral bioavailability of scutellarin. The gender differences of pharmacokinetic parameters of scutellarin and scutellarein are due to the higher CL(int) and lower absorption in male rats.
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Apigenina/farmacocinética , Cromatografía Líquida de Alta Presión/métodos , Erigeron/química , Glucuronatos/farmacocinética , Administración Oral , Animales , Apigenina/administración & dosificación , Área Bajo la Curva , Disponibilidad Biológica , Heces , Femenino , Tracto Gastrointestinal/metabolismo , Glucuronatos/administración & dosificación , Semivida , Inyecciones Intravenosas , Masculino , Microsomas Hepáticos/metabolismo , Ratas , Ratas Sprague-Dawley , Factores SexualesRESUMEN
Pulmonary haemorrhage is an important complication of leptospirosis. We herein report an uncommon case of severe pulmonary haemorrhage and multiple organ failure caused by leptospirosis in a 49-year-old man who was previously healthy. He was a farm worker who was admitted to the hospital because of haemoptysis. He had worked in a paddy field 4 days prior to admission. Chest computed tomography revealed pulmonary haemorrhage, which rapidly deteriorated into haemorrhagic shock and multiple organ failure. Based on the patient's possible history of contact with contaminated water and the DNA sequence of Leptospira detected in his bronchoalveolar lavage fluid, the patient was diagnosed with pulmonary haemorrhagic leptospirosis. Despite the administration of a fluid bolus, norepinephrine, broad-spectrum antibiotics, and haemostatics, and even with administration of a blood transfusion and extracorporeal life support, the pulmonary haemorrhage could not be controlled effectively. The patient eventually died of haemorrhagic shock. Leptospirosis can be a life-threatening disease despite aggressive treatment, even with extracorporeal life support. Next-generation sequencing can provide important diagnostic clues for patients with atypical leptospirotic symptoms.
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Leptospira , Leptospirosis , Enfermedades Pulmonares , Hemorragia/diagnóstico por imagen , Hemorragia/etiología , Humanos , Leptospirosis/complicaciones , Leptospirosis/diagnóstico , Enfermedades Pulmonares/complicaciones , Enfermedades Pulmonares/diagnóstico por imagen , Masculino , Persona de Mediana Edad , Insuficiencia Multiorgánica/etiologíaRESUMEN
Previously we reported the nucleotide sequence of a 14-3-3 cDNA cloned from the unicellular green alga Dunaliella salina, however, the nucleotide sequence of this gene have not been reported so far. In the present study, the cloning and characterization of the nucleotide sequence, the gene copy and expression were undertaken. The coding sequence of the gene was found to be interrupted by five introns of 132, 266, 153, 152 and 625 bp, respectively. Introns 3-5 were found in conserved positions as compared to the Chlamydomonas reinhardtii 14-3-3 gene. D. salina 14-3-3 cDNA was inserted into the prokaryotic expression plasmid pET-28 and transformed into E. coli BL21, and the recombinant expressed 14-3-3 protein was purified from E. coli and immunized the rabbit. Indirect ELISA coated with 14-3-3 illustrated that the rabbit antisera titration was 1:1.00E + 06. Western blotting assays confirmed that prepared rabbit antibodies could recognize the recombinant 14-3-3 protein. Southern blotting results showed that there was only one copy of the 14-3-3 present in the genome of D. salina and 14-3-3 expression did not change throughout the Dnualiella cell cycle.
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Proteínas 14-3-3/genética , Proteínas 14-3-3/metabolismo , Chlorophyta/genética , Chlorophyta/metabolismo , Proteínas 14-3-3/inmunología , Animales , Secuencia de Bases , Clonación Molecular , Expresión Génica , Perfilación de la Expresión Génica , Nucleótidos/análisis , Conejos , Tolerancia a la Sal/genética , Análisis de Secuencia de ADNRESUMEN
OBJECTIVE: To investigate the absorption and transepithelial transport characteristics of scutellarin and scutellarein in the human colonic adenocarcinoma cell (Caco-2) monolayer model. The influence factors on these two compounds' absorption were investigated, such as buffer solution, duration of culture, and inhibitors of multidrug resistance-associated protein 2 (MRP(2)), breast cancer drug resistance protein (BCRP) and P-glycoprotein (P-gp). METHODS: By using Caco-2 monolayer as an intestinal epithelial cell model, the transport process was studied from apical (AP) side to basolateral (BL) side or from BL to AP. The two compounds were determined by high-performance liquid chromatography coupled with diode-array-detector detection. Transport parameters and apparent permeability coeffients (P(app)) were calculated. RESULTS: The P(app) values of scutellarin and scutellarein were different in two buffer solutions, respectively. In phosphate buffered saline, scutellarin had no absorption from AP to BL, while its P(app) value was 0.74×10(-6) to 1.58×10(-6) cm/s from BL to AP. The P(app) values of scutellarein were 4.33×10(-6) to 6.79×10(-6) cm/s and 1.32×10(-6) to 2.56×10(-6) cm/s from AP to BL and from BL to AP, respectively. The P(app) value gradually decreased with time. The absorption of scutellarein was better than that of scutellarin. The scutellarin absorption was improved by verapamil, MK-571 and reserpine. The scutellarein absorption was improved by verapamil whereas its excretion was improved by MK-571. CONCLUSION: Absorption of scutellarin is difficult in Caco-2 monolayer cells, which contributes to its low bioavailability. Scutellarein absorption is better than scutellarin absorption. Scutellarein transepithelial transport is passive diffusion. The inhibitor of P-gp can improve scutellarin and scutellarein transportation. The inhibitors of MRP(2) and BCRP can promote transportation of scutellarin. The inhibitor of MRP(2) can promote efflux of scutellarein. The multidrug resistance-associated protein may be the second reason for low bioavailability of scutellarin.
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Adenocarcinoma/patología , Apigenina/farmacocinética , Neoplasias del Colon/patología , Glucuronatos/farmacocinética , Transporte Biológico , Células CACO-2 , Humanos , Proteína 2 Asociada a Resistencia a Múltiples MedicamentosRESUMEN
Inflammation is a leading cause of severe acute pancreatitis (SAP). MicroRNAs (miRNAs) are emerging as important regulators involved in the pathogenesis of many diseases including pancreatitis. To identify miRNAs that contribute to the pathology of SAP, we carried out a miRNA-specific microarray analysis using the biopsies donated by SAP patients. We totally obtained 50 differentially expressed miRNAs, including 20 upregulated and 30 downregulated miRNAs, respectively. We focused our current study on revealing the downstream target and the upstream regulatory mechanism of miR-589-5p, the most downregulated miRNA in our candidate lists. Our prediction results indicated that miR-589-5p might target TRAF6 (tumor necrosis factor receptor-associated factor 6), a critical member of the TLR4/NF-kB (Toll-like receptor 4/nuclear transcription factor-kB) pathway. Using different strategies such as in vitro overexpression or downregulation of miR-589-5p and treatment with lipopolysaccharide (LPS), we found that the expression of TRAF6 was regulated by two-layer mechanisms. On the one hand, TRAF6 was transcriptionally controlled by a DNA methylation mediated downregulation of miR-589-5p. On the other hand, the activation of TLR4/NF-kB signaling also could increase the protein level of TRAF6. The increased TRAF6 aggravated the downstream signaling and caused the translocation of NF-kB subunits from the cytoplasm to the nucleus, where NF-kB transcription factors induced the expression of proinflammatory cytokine genes. The maturation and production of proinflammatory cytokines induced inflammatory response and caused the occurrence of SAP.
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Monocytes and macrophages are the two major cell types involved in innate immunity. Exosomes act as signaling molecules to regulate cell-to-cell communication by releasing proteins, mRNAs, microRNAs (miRNAs), and long noncoding RNAs (lncRNAs). However, it is still unclear whether monocyte-derived exosomes are involved in the communication between monocytes and macrophages. In this study, we analyzed the differentially expressed lncRNA profiles in monocytes isolated from blood samples of healthy controls and acute lung injury (ALI) patients. We focused our study on investigating the signaling downstream of CLMAT3 (colorectal liver metastasis-associated transcript 3), a lncRNA that regulated proinflammatory cytokine genes. We revealed that CLMAT3 specifically targeted CtBP2 (C-terminal binding protein 2) and repressed its expression. Elevated CtBP2 acted as a coactivator to assemble a transcriptional complex with histone acetyltransferase p300 and NF-κB (nuclear factor κB) subunits. In vitro coculture and in vivo injection of ALI monocyte-derived exosomes increased the production of proinflammatory cytokines. Importantly, the administration of two CtBP2 inhibitors, NSC95397 and MTOB, could significantly reverse CtBP2-mediated transactivation. Collectively, our results support a model in which monocyte-derived exosomal CLMAT3 activates the CtBP2-p300-NF-κB complex to induce proinflammatory cytokines, thus contributing to the pathogenesis of ALI.
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Emerging evidence indicates that microRNAs (miRNAs) play fundamental roles in the pathogenesis of multiple diseases, including acute lung injury (ALI). Here, we discovered that miR-199a-3p was significantly downregulated in ALI lung tissues using a microarray analysis. In vitro lipopolysaccharide (LPS) treatment of the human epithelial cell line A549 and the human macrophage cell line U937 caused a decrease of miR-199a-3p. Mechanically, miR-199a-3p specifically bound to the 3'-untranslated region (3'-UTR) of NLRP1 (nucleotide-binding oligomerization domain, leucine-rich repeat and pyrin domain-containing protein 1), a critical member of inflammasomes. Ectopic overexpression or downregulation of miR-199a-3p resulted in the repression or induction of NLRP1, respectively, thereby downregulating or activating its downstream events. Moreover, transcription factor FOXP3 (forkhead box P3) was able to specifically bind to the promoter of miR-199a-3p. Knockdown or overexpression of FOXP3 resulted in a decrease or induction miR-199a-3p expression, respectively. Using immunoprecipitation (IP), mass spectrometry and co-IP assays, we found that FOXP3 formed a transcriptional complex with HDAC1 (histone deacetylase 1) and CtBP2 (C-terminal-binding protein 2). Collectively, our results suggested that the CtBP2-HDAC1-FOXP3 transcriptional complex (CHFTC) could specifically bind to the promoter of miR-199a-3p and repress its expression. Downregulation of miR-199a-3p eliminated its inhibition of NLRP1, causing activation of NLRP1 and cleavage of pro-IL-1ß and pro-IL-18 mediated by Caspase-1. The secretion of IL-1ß and IL-18 further aggravated the inflammatory response and resulted in the occurrence of ALI.
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Lesión Pulmonar Aguda/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Lipopolisacáridos/toxicidad , MicroARNs/metabolismo , Células A549 , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Proteínas Reguladoras de la Apoptosis/genética , Proteínas Reguladoras de la Apoptosis/metabolismo , Citocinas/genética , Citocinas/metabolismo , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Humanos , MicroARNs/genética , Proteínas NLR , Regiones Promotoras Genéticas , Unión Proteica , Células U937RESUMEN
PURPOSE: The purpose of this study was to retrospectively evaluate the efficacy and safety of side-hole catheter technique for transarterial chemoembolization (TACE) via transradial artery access (TRA) in patients with hepatocellular carcinoma. MATERIALS AND METHODS: From November 2015 to August 2017, a total of 1040 TACE procedures were performed via TRA for hepatocellular carcinoma. In 10 (1%) of these 1040 TACE procedures via TRA, conventional microcatheter technique (CMT) failed and side-hole catheter technique was attempted. RESULTS: Ten procedures of selective catheterizations by CMT failed due to the poor stability of the angiographic catheters or the target artery arising from the very proximal portion of the parent artery. These arteries included the right inferior phrenic artery in eight patients, one left gastric artery, and one right renal capsular artery. Cobra or MPA catheter with the microcatheter through the side-hole yielded a technical success rate of 100%. No procedure-related complications were observed. The mean time required to catheterize the target artery with the side-hole catheter was 9.5 min (5-15 min). CONCLUSION: Side-hole catheter technique may enable the completion of chemoembolization in cases that a potential tumor-feeding vessel cannot be catheterized by means of CMT for TACE via TRA.
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Carcinoma Hepatocelular/terapia , Cateterismo/instrumentación , Cateterismo/métodos , Quimioembolización Terapéutica/instrumentación , Quimioembolización Terapéutica/métodos , Neoplasias Hepáticas/terapia , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , Resultado del TratamientoRESUMEN
Most traditional post-electrophoretic processes need several hours to several days to finish the whole staining process and traditional staining solutions all contain methanol, acetic acid, or phosphoric acid, which not only produce the unpleasant smell but also cause environmental pollution. Here a fixation-free, fast protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis using Coomassie blue is described. The protocol includes only staining and quick washing steps, can be completed in 0.5 h. It has a sensitivity of 10 ng. In addition, the dye stain does not contain any acid or methanol.