RESUMO
High-performance metabolic analysis is emerging in the diagnosis and prognosis of breast cancer (BrCa). Still, advanced tools are in demand to deliver the application potentials of metabolic analysis. Here, we used fast nanoparticle-enhanced laser desorption/ionization mass spectrometry (NPELDI-MS) to record serum metabolic fingerprints (SMFs) of BrCa in seconds, achieving high reproducibility and low consumption of direct serum detection without treatment. Subsequently, machine learning of SMFs generated by NPELDI-MS functioned as an efficient readout to distinguish BrCa from non-BrCa with an area under the curve of 0.948. Furthermore, a metabolic prognosis scoring system was constructed using SMFs with effective prediction performance toward BrCa (P < 0.005). Finally, we identified a biomarker panel of seven metabolites that were differentially enriched in BrCa serum and their related pathways. Together, our findings provide an efficient serum metabolic tool to characterize BrCa and highlight certain metabolic signatures as potential diagnostic and prognostic factors of diseases including but not limited to BrCa.
Assuntos
Neoplasias da Mama , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/metabolismo , Feminino , Humanos , Espectrometria de Massas/métodos , Prognóstico , Reprodutibilidade dos TestesRESUMO
Historical marquisette curtains were composed of lightweight fabrics, woven in an open-mesh and leno-type weave, usually made of silk, and found in Qing imperial buildings. As panel curtains, they were exposed to light, and so underwent fading. This study investigated the manufacturing technology and fading mechanism of dyed marquisette fabric from the Studio of Cleansing Fragrance, the Palace Museum (Beijing). The technological aspects were identified. The types of weave, fiber, and adhesive used to fix the curtain to the wooden frame were identified through microscopic observation and infrared spectroscopy. A color change characterization was performed based on UV-visible diffuse reflectance spectra. The textile colorant was identified as malachite green (MG), and its degradation by light was subsequently studied by dynamic photolysis experiments in a kinetic solution for the rapid exploration of by-products. The main degradation pathways were thus identified and the factors responsible for the induced color changes were discussed. A comparison of the liquid chromatography-mass spectrometry (LC-MS) results of the products derived from the photolysis method as well as of the samples extracted from the object allowed for the identification of the presence of different degradation pathways in the faded and unfaded parts of the textile. A metabolomics analysis was applied to account for the differences in the degradation pathways.
Assuntos
Corantes , Perfumes , Pequim , Corantes/química , Museus , Odorantes , Corantes de RosanilinaAssuntos
Braço , Humanos , Feminino , Braço/patologia , Diagnóstico Diferencial , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Clinically, depigmentation after local corticosteroid injection is not rare. But there are less articles about its reflectance confocal microscopy (RCM) and histological features. This study aimed to define the RCM features and histopathologic findings of hypopigmentation after local corticosteroid injection and to analyze the correlations between the above two methods. METHODS: Forty cases with hypopigmentation after local corticosteroid injection were used to analyze the clinical and RCM features. Subsequently, for 20 of 40, an excision biopsy of the same imaged areas for histopathologic examination was executed. RESULTS: Our results showed that all 40 cases had round or ellipse hypopigmented macules with obscure boundary and 26 of 40 lesions' long diameter went along limbs. The RCM features and the histological findings revealed all patients had variable degrees of epidermal thinning, flattening rete ridges, reduced melanin, and no inflammatory cell infiltration. MART-1 analysis revealed the number of melanocytes was normal but with no or less melanin by Fontana-Masson staining. CONCLUSIONS: Depigmentation after local corticosteroid injection was a kind of disease with intact melanocytes, whose function was impaired. RCM features offer a high consistency with histopathologic findings. It thus constitutes a promising adjuvant tool for its diagnosis and for therapeutic follow-up.
Assuntos
Corticosteroides/efeitos adversos , Hipopigmentação , Microscopia Confocal/métodos , Pele , Corticosteroides/administração & dosagem , Adulto , Idoso , Feminino , Histocitoquímica , Humanos , Hipopigmentação/induzido quimicamente , Hipopigmentação/diagnóstico por imagem , Hipopigmentação/patologia , Injeções Intradérmicas/efeitos adversos , Masculino , Pessoa de Meia-Idade , Pele/química , Pele/diagnóstico por imagem , Pele/patologiaRESUMO
Glycosylation, which is catalyzed by UDP-glycosyltransferases (UGTs), is an important biological modification for the structural and functional diversity of ginsenosides. In this study, the promiscuous UGT109A1 from Bacillus subtilis was used to synthesize unnatural ginsenosides from natural ginsenosides. UGT109A1 was heterologously expressed in Escherichia coli and then purified by Ni-NTA affinity chromatography. Ginsenosides Re, Rf, Rh1, and R1 were selected as the substrates to produce the corresponding derivatives by the recombinant UGT109A1. The results showed that UGT109A1 could transfer a glucosyl moiety to C3-OH of ginsenosides Re and R1, and C3-OH and C12-OH of ginsenosides Rf and Rh1, respectively, to produce unnatural ginsenosides 3,20-di-O-ß-d-glucopyranosyl-6-O-[α-l-rhamnopyrano-(1â2)-ß-d-glucopyranosyl]-dammar-24-ene-3ß,6α,12ß,20S-tetraol (1), 3,20-di-O-ß-d-glucopyranosyl-6-O-[ß-d-xylopyranosyl-(1â2)-ß-d-glucopyranosyl]-dammar-24-ene-3ß,6α,12ß,20S-tetraol (6), 3-O-ß-d-glucopyranosyl-6-O-[ß-d-glucopyranosyl-(1â2)-ß-d-glucopyranosyl]-dammar-24-ene-3ß,6α,12ß,20S-tetraol (3), 3,12-di-O-ß-d-glucopyranosyl-6-O-[ß-d-glucopyranosyl-(1â2)-ß-d-glucopyranosyl]-dammar-24-ene-3ß,6α,12ß,20S-tetraol (2), 3,6-di-O-ß-d-glucopyranosyl-dammar-24-ene-3ß,6α,12ß,20S-tetraol (5), and 3,6,12-tri-O-ß-d-glucopyranosyl-dammar-24-ene-3ß,6α,12ß,20S-tetraol (4). Among the above products, 1, 2, 3, and 6 are new compounds. The maximal activity of UGT109A1 was achieved at the temperature of 40 °C, in the pH range of 8.0â»10.0. The activity of UGT109A1 was considerably enhanced by Mg2+, Mn2+, and Ca2+, but was obviously reduced by Cu2+, Co2+, and Zn2+. The study demonstrated that UGT109A1 was effective in producing a series of unnatural ginsenosides through enzymatic reactions, which could pave a way to generate promising leads for new drug discovery.
Assuntos
Bacillus subtilis/enzimologia , Ginsenosídeos/síntese química , Glucosiltransferases/química , Bacillus subtilis/genética , Cromatografia Líquida de Alta Pressão , Ativação Enzimática , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Ginsenosídeos/química , Glucosiltransferases/genética , Glucosiltransferases/isolamento & purificação , Glicosilação , Concentração de Íons de Hidrogênio , Técnicas In Vitro , Íons/química , Metais/química , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/isolamento & purificaçãoRESUMO
As the main bioactive constituents of Panax species, ginsenosides possess a wide range of notable medicinal effects such as anti-cancer, anti-oxidative, antiaging, anti-inflammatory, anti-apoptotic and neuroprotective activities. However, the increasing medical demand for ginsenosides cannot be met due to the limited resource of Panax species and the low contents of ginsenosides. In recent years, biotechnological approaches have been utilized to increase the production of ginsenosides by regulating the key enzymes of ginsenoside biosynthesis, while synthetic biology strategies have been adopted to produce ginsenosides by introducing these genes into yeast. This review summarizes the latest research progress on cloning and functional characterization of key genes dedicated to the production of ginsenosides, which not only lays the foundation for their application in plant engineering, but also provides the building blocks for the production of ginsenosides by synthetic biology.
Assuntos
Ginsenosídeos/biossíntese , Alquil e Aril Transferases/genética , Alquil e Aril Transferases/metabolismo , Vias Biossintéticas/genética , Clonagem Molecular , Sistema Enzimático do Citocromo P-450/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Farnesil-Difosfato Farnesiltransferase/genética , Farnesil-Difosfato Farnesiltransferase/metabolismo , Ginsenosídeos/química , Glicosiltransferases/genética , Glicosiltransferases/metabolismo , Hidroximetilglutaril-CoA-Redutases NADP-Dependentes/genética , Hidroximetilglutaril-CoA-Redutases NADP-Dependentes/metabolismo , Transferases Intramoleculares/genética , Transferases Intramoleculares/metabolismo , Esqualeno Mono-Oxigenase/genética , Esqualeno Mono-Oxigenase/metabolismoRESUMO
Pyrolysis-gas chromatography/mass spectrometry (Py-GC/MS) is suitable for the qualitative analysis of natural organic materials in cultural relics owing to its ability of accurately and rapidly identifying trace quantities of organic components in samples. In the present study, pyrolytic components of starch and peach gum, which are commonly used in ancient Chinese cultural relics such as calligraphic works, paintings, architecture, and objects, as well as gum Arabic, which is commonly used in western cultural relics, were systematically characterized using Py-GC/MS. As polysaccharide materials are often found mixed with other organic materials in cultural relics, an online methylation Py-GC/MS procedure previously established by the authors' research group, which is suitable for the qualitative analysis of drying oils, proteins, and waxes, was performed to analyze the starch, peach gum, and gum Arabic reference samples. The established experimental procedure can be used to comprehensively analyze the polysaccharides and other organic materials in cultural relics owing to its slow heating rate, long running time, wide mass spectrometry detection range, and high chromatographic separation efficiency. The experimental results were examined by dividing the pyrolytic products into three sections depending on the retention time. In the first section (2.5-10 min), pyrolytic products from the three sample types were similar and mostly included small molecule furans and ketones. However, the peach gum and gum Arabic contain a wider variety of pyrolytic products than starch, possibly because starch contains only glucose, whereas peach gum and gum Arabic contain a variety of monosaccharides. In the middle section (10-19 min), the pyrolytic products of starch, peach gum, and gum Arabic mostly consisted of furanones and other components. However, a unique profile was produced for each material type. Nevertheless, it should be noted that 2-methoxy-phenol and maltol were detected in all three materials with high chromatographic peak intensities. Therefore, these components are suitable markers for the identification of polysaccharides. In the last section (19-60 min), a variety of monosaccharide derivatives and monosaccharide oligomer derivatives were detected in the reference samples. The pyrolytic products of peach gum and gum Arabic were similar but completely different from those of starch. Therefore, starch, peach gum, and gum Arabic could be distinguished according to their pyrolytic products. In particular, 1,6-anhydro-ß-D-glucopyranose was detected in starch with an extremely high intensity and was undetected in either of the other reference materials. Thus, this compound could act as a characteristic pyrolytic component for the identification of starch. Peach gum and gum Arabic could be rapidly distinguished according to the extracted ion chromatograms for m/z 60 and m/z 101, which represented major fragments of the pyrolytic products of peach gum and gum Arabic in the last section of the chromatogram. The established Py-GC/MS method was successfully applied to the identification of starch in binding materials from the rim of a globular carved red lacquer vase with dragon and cloud motifs, as well as in paper collected from a Tieluo with Bingdihanfang written by Min-ning. Both objects are part of the Palace Museum collection and originate from the Qing Dynasty. Research results of the present work are easy to be popularized. This study provides a method suitable for the accurate and rapid identification of polysaccharide materials in cultural relics, as well as a scientific basis for the research, conservation, and restoration of similar cultural relics. However, it should be noted that aging and the presence of other organic or inorganic materials in cultural relics may interfere with the detection of polysaccharide materials. Therefore, a further investigation on the aging behavior of polysaccharides and the effects of other materials on the identification of polysaccharides is required.
Assuntos
Goma Arábica , Pirólise , Cromatografia Gasosa-Espectrometria de Massas/métodos , Goma Arábica/análise , Monossacarídeos/análise , Polissacarídeos/análise , AmidoRESUMO
OBJECTIVE: To investigate the mRNA and proten expression of coxsackievirus and adenovirus receptor (CAR) in the corresponding normal lung tissue, para-neoplastic tissue and lung cancer tissue, and the correlation of CAR expression with the carcinogenesis as well as the expression difference in various clinicopathologic parameters. METHODS: The expression of CAR mRNA and protein in the samples from 32 lung cancer patients was determined by RT-PCR and Western blot, respectively. RESULTS: The expression level of CAR mRNA and protein in normal lung tissue, paraneoplastic tissue and cancer tissue were 1.000 +/- 0.012, 1.048 +/- 0.035, 1.282 +/- 0.072, and 0.902 +/- 0.038, 0.944 +/- 0.042, 1.08 +/- 0.052, respectively, with a statistical significance among the groups (P = 0.022, P = 0.007, P = 0.009, P = 0.027). There was a statistically significant positive correlation between expression of CAR mRNA and that of CAR protein (r = 0.448, P = 0.026). The expression levels of CAR were significantly different among different pathological types (P = 0.012), with a high level of CAR in all 7 bronchiolo-alveolar carcinoma (BAC, P = 0.029). However, there was no statistical significance in other clinicopathologic parameters (P > 0.05), including gender, age, smoking or not, tumor size, with or without lymph node metastasis and TNM stage. CONCLUSION: The expression of CAR mRNA and protein in cancer tissue samples are significantly higher than that in the normal and paraneoplastic samples, indicating that CAR might play a crucial role in the carcinogenesis. It may become a new potential prognostic marker for lung cancer patients.
Assuntos
Adenocarcinoma Bronquioloalveolar/metabolismo , Carcinoma Pulmonar de Células não Pequenas/metabolismo , Neoplasias Pulmonares/metabolismo , Receptores Virais/metabolismo , Adenocarcinoma Bronquioloalveolar/patologia , Adulto , Idoso , Biomarcadores Tumorais/metabolismo , Western Blotting , Carcinoma Pulmonar de Células não Pequenas/patologia , Proteína de Membrana Semelhante a Receptor de Coxsackie e Adenovirus , Feminino , Humanos , Pulmão/metabolismo , Neoplasias Pulmonares/patologia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Prognóstico , RNA Mensageiro/metabolismo , Receptores Virais/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
PURPOSE: To investigate the differences in postoperative sore throat and hoarseness by adjustment of endotracheal tube cuff pressure (CP) during nitrous oxide (N2O) and air anesthesia. METHODS: A one-equivalent control group pretest-posttest design was used. Data were collected from August 8 to October 19, 2017 and analyzed using the independent t-test and repeated measures ANOVA. Eighty-four participants were enrolled and divided into three groups: 28 in the Control Group (CP adjusted every 30 minutes using N2O), 28 in Experimental Group 1 (CP adjusted every 10 minutes using N2O), and 28 in Experimental Group 2 (non-adjusted CP using air), all of whom underwent urologic, gynecologic, and orthopedic surgeries at the G University hospital. Sore throat was assessed using a numeric rating scale; hoarseness was evaluate using the Stout classification at 1, 6, and 24 hours after surgery. RESULTS: Scores for sore throat and hoarseness were significantly different between the groups at each measurement time, and scores were consistently higher in the control group. During subsequent measurements, sore throat and hoarseness scores were significantly lower at 6 hours. Cuff pressure changed significantly using air anesthesia (χ²=10.41, p=.015) up to 2 hours after induction. Severe sore throat and hoarseness was observed for up to 6 hours after surgery. CONCLUSION: Cuff pressure adjustment at short time intervals would be helpful in reducing postoperative sore throat and hoarseness. Nursing intervention focused on prevention of sore throat and hoarseness should be required up to 6 hours postoperatively in patients undergoing endotracheal intubation.
Assuntos
Anestesia Geral/métodos , Rouquidão/etiologia , Óxido Nitroso/química , Faringite/etiologia , Idoso , Anestesia Geral/efeitos adversos , Feminino , Procedimentos Cirúrgicos em Ginecologia , Humanos , Masculino , Pessoa de Meia-Idade , Procedimentos Ortopédicos , Complicações Pós-Operatórias , PressãoRESUMO
BACKGROUND: As the number of patients with diffuse coronary artery disease continues to grow, there is renewed interest in alternative methods of perfusing the ischemic myocardium. We tested the feasibility of myocardial retroperfusion via a direct left ventricle-to-great cardiac vein (LV-GCV) conduit to support regional contractility in this setting. METHODS: LV-GCV flow was established using an extracorporeal circuit in 5 dogs. Left ventricle (LV) pressure, aortic pressure, regional myocardial segment length, and circuit blood flow were measured prior to left anterior descending coronary artery (LAD) ligation, following LAD ligation, and after LV-GCV circuit placement. To eliminate backward flow during diastole, an in-line flow regulator was placed. Regional myocardial function was quantified by pressure-segment length loop area divided by end-diastolic segment length (PSLA/EDSL). RESULTS: LAD ligation reduced PSLA/EDSL from 10.0 +/- 1.2 mm Hg mm to 1.6 +/- 0.3 mm Hg mm (P < .05). With LV-GCV retroperfusion, mean peak systolic flow was +152 +/- 14 mL/min, mean peak diastolic flow was -39 +/- 11 mL/min, and net mean flow was +36 +/- 13 mL/min. Regional function recovered to approximately 39% of baseline (3.9 +/- 0.4 mm Hg mm, P < .05). Upon elimination of backflow, mean flow increased to +41 +/- 12 mL/min and regional function recovered even further to approximately 47% of baseline (4.6 +/- 0.7 mm Hg mm, P < .05). CONCLUSIONS: A LV-GCV circuit can significantly restore regional function to the acutely ischemic myocardium. An inline valve that eliminates backward diastolic flow improves regional function even further. This approach may provide an effective therapy for diffuse coronary disease not amenable to traditional revascularization strategies.
Assuntos
Anastomose Cirúrgica/métodos , Vasos Coronários/cirurgia , Ventrículos do Coração/cirurgia , Isquemia Miocárdica/cirurgia , Revascularização Miocárdica/métodos , Veias/cirurgia , Disfunção Ventricular Esquerda/cirurgia , Animais , Cães , Isquemia Miocárdica/complicações , Resultado do Tratamento , Disfunção Ventricular Esquerda/etiologiaRESUMO
The cardiac sarcoplasmic reticulum calcium-ATPase (SERCA2a), Na+/Ca2+ exchanger (NCX1), and ryanodine receptor (RyR2) are proteins involved in the regulation of myocyte calcium. We tested whether exercise training (ET) alters those proteins during development of chronic heart failure (CHF). Ten dogs were chronically instrumented to permit hemodynamic measurements. Five dogs underwent 4 wk of cardiac pacing (210 beats/min for 3 wk and 240 beats/min for the 4th wk), whereas five dogs underwent the same pacing regimen plus daily ET (5.1 +/- 0.3 km/h, 2 h/day). Paced animals developed CHF characterized by hemodynamic abnormalities and reduced ejection fraction. ET preserved resting hemodynamics and ejection fraction. Left ventricular samples were obtained from all dogs and another five normal dogs for mRNA (Northern analysis, band intensities normalized to glyceraldehyde-3-phosphate dehydrogenase) and protein level (Western analysis, band intensities normalized to tubulin) measurements. In failing hearts, SERCA2a was decreased by 33% (P < 0.05) and 65% (P < 0.05) in mRNA and protein level, respectively, compared with normal hearts; there was only an 8.6% reduction in mRNA and a 32% reduction in protein in exercised animals (P < 0.05 from CHF). mRNA expression of NCX1 increased by 44% in paced-only dogs compared with normal (P < 0.05) but only by 22% in trained dogs (P < 0.05 vs. CHF); protein level of NCX1 was elevated in paced-only dogs (71%, P < 0.05) but partially normalized by ET (33%, P < 0.05 from CHF). RyR2 was not altered in any of the dogs. In conclusion, long-term ET may ameliorate cardiac deterioration during development of CHF, in part via normalization of myocardial calcium-handling proteins.