RESUMEN
The biology-material hybrid method for chemical-electricity conversion via microbial fuel cells (MFCs) has garnered significant attention in addressing global energy and environmental challenges. However, the efficiency of these systems remains unsatisfactory due to the complex manufacturing process and limited biocompatibility. To overcome these challenges, here, we developed a simple bio-inorganic hybrid system for bioelectricity generation in Shewanella oneidensis (S. oneidensis) MR-1. A biocompatible surface display approach was designed, and silver-binding peptide AgBP2 was expressed on the cell surface. Notably, the engineered Shewanella showed a higher electrochemical sensitivity to Ag+, and a 60 % increase in power density was achieved even at a low concentration of 10 µM Ag+. Further analysis revealed significant upregulations of cell surface negative charge intensity, ATP metabolism, and reducing equivalent (NADH/NAD+) ratio in the engineered S. oneidensis-Ag nanoparticles biohybrid. This work not only provides a novel insight for electrochemical biosensors to detect metal ions, but also offers an alternative biocompatible surface display approach by combining compatible biomaterials with electricity-converting bacteria for advancements in biohybrid MFCs.
Asunto(s)
Fuentes de Energía Bioeléctrica , Técnicas Biosensibles , Shewanella , Plata , Shewanella/metabolismo , Shewanella/química , Fuentes de Energía Bioeléctrica/microbiología , Técnicas Biosensibles/métodos , Plata/química , Materiales Biocompatibles/química , Nanopartículas del Metal/química , Electricidad , Técnicas Electroquímicas/métodosAsunto(s)
Insuficiencia Cardíaca , Guías de Práctica Clínica como Asunto , Inhibidores del Cotransportador de Sodio-Glucosa 2 , Volumen Sistólico , Humanos , Insuficiencia Cardíaca/tratamiento farmacológico , Insuficiencia Cardíaca/fisiopatología , Volumen Sistólico/fisiología , Inhibidores del Cotransportador de Sodio-Glucosa 2/uso terapéutico , Sistema de Registros , Suecia/epidemiologíaRESUMEN
The objective of this study was to isolate and characterize collagen and angiotensin-I-converting enzyme (ACE)-inhibitory (ACEi) peptides from the swim bladders of monkfish (Lophius litulon). Therefore, acid-soluble collagen (ASC-M) and pepsin-soluble collagen (PSC-M) with yields of 4.27 ± 0.22% and 9.54 ± 0.51%, respectively, were extracted from monkfish swim bladders using acid and enzyme methods. The ASC-M and PSC-M contained Gly (325.2 and 314.9 residues/1000 residues, respectively) as the major amino acid, but they had low imino acid content (192.5 and 188.6 residues/1000 residues, respectively) in comparison with collagen from calf skins (CSC) (216.6 residues/1000 residues). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns and ultraviolet (UV) absorption spectrums of ASC-M and PSC-M illustrated that they were mainly composed of type I collagen. Subsequently, three ACEi peptides were isolated from a PSC-M hydrolysate prepared via a double-enzyme system (alcalase + neutrase) and identified as SEGPK (MHP6), FDGPY (MHP7) and SPGPW (MHP9), with molecular weights of 516.5, 597.6 and 542.6 Da, respectively. SEGPK, FDGPY and SPGPW displayed remarkable anti-ACE activity, with IC50 values of 0.63, 0.94 and 0.71 mg/mL, respectively. Additionally, a molecular docking assay demonstrated that the affinities of SEGPK, FDGPY and SPGPW with ACE were -7.3, -10.9 and -9.4 kcal/mol, respectively. The remarkable ACEi activity of SEGPK, FDGPY and SPGPW was due to their connection with the active pockets and/or sites of ACE via hydrogen bonding, hydrophobic interaction and electrostatic force. Moreover, SEGPK, FDGPY and SPGPW could protect HUVECs by controlling levels of nitric oxide (NO) and endothelin-1 (ET-1). Therefore, this work provides an effective means for the preparation of collagens and novel ACEi peptides from monkfish swim bladders, and the prepared ACEi peptides, including SEGPK, FDGPY and SPGPW, could serve as natural functional components in the development of health care products to control hypertension.
Asunto(s)
Colágeno , Peptidil-Dipeptidasa A , Animales , Simulación del Acoplamiento Molecular , Colágeno/química , Peces/metabolismo , Péptidos/farmacología , Péptidos/química , Ácidos/química , AngiotensinasRESUMEN
Following a DNA double strand break (DSB), several nucleases and helicases coordinate to generate single-stranded DNA (ssDNA) with 3' free ends, facilitating precise DNA repair by homologous recombination (HR). The same nucleases can act on stalled replication forks, promoting nascent DNA degradation and fork instability. Interestingly, some HR factors, such as CtIP and BRCA1, have opposite regulatory effects on the two processes, promoting end resection at DSB but inhibiting the degradation of nascent DNA on stalled forks. However, the reason why nuclease actions are regulated by different mechanisms in two DNA metabolism is poorly understood. We show that human HELQ acts as a DNA end resection regulator, with opposing activities on DNA end resection at DSBs and on stalled forks as seen for other regulators. Mechanistically, HELQ helicase activity is required for EXO1-mediated DSB end resection, while ssDNA-binding capacity of HELQ is required for its recruitment to stalled forks, facilitating fork protection and preventing chromosome aberrations caused by replication stress. Here, HELQ synergizes with CtIP but not BRCA1 or BRCA2 to protect stalled forks. These findings reveal an unanticipated role of HELQ in regulating DNA end resection at DSB and stalled forks, which is important for maintaining genome stability.
Asunto(s)
Roturas del ADN de Doble Cadena , Replicación del ADN , Humanos , ADN Helicasas/genética , Reparación del ADN , Recombinación Homóloga/genéticaRESUMEN
The DNA damage response (DDR) system plays an important role in maintaining genome stability and preventing related diseases. The DDR network comprises many proteins and posttranslational modifications (PTMs) to proteins, which work in a coordinated manner to counteract various genotoxic stresses. Lysine crotonylation (Kcr) is a newly identified PTM occurring in both core histone and non-histone proteins in various organisms. This novel PTM is classified as a reversible acylation modification, which is regulated by a variety of acylases and deacylases and the intracellular crotonyl-CoA substrate concentration. Recent studies suggest that Kcr links cellular metabolism with gene regulation and is involved in numerous cellular processes. In this review, we summarize the regulatory mechanisms of Kcr and its functions in DDR, including its involvement in double-strand break (DSB)-induced transcriptional repression, DSB repair, and the DNA replication stress response.
Asunto(s)
Histonas , Lisina , Lisina/química , Histonas/metabolismo , Procesamiento Proteico-Postraduccional , Reparación del ADN , Daño del ADNRESUMEN
The reversible post-translational modification (PTM) of proteins plays an important role in many cellular processes. Lysine crotonylation (Kcr) is a newly identified PTM, but its functional significance remains unclear. Here, we found that Kcr is involved in the replication stress response. We show that crotonylation of histone H2A at lysine 119 (H2AK119) and ubiquitination of H2AK119 are reversibly regulated by replication stress. Decrotonylation of H2AK119 by SIRT1 is a prerequisite for subsequent ubiquitination of H2AK119 by BMI1. Accumulation of ubiquitinated H2AK119 at reversed replication forks leads to the release of RNA Polymerase II and transcription repression in the vicinity of stalled replication forks. These effects attenuate transcription-replication conflicts (TRCs) and TRC-associated R-loop formation and DNA double-strand breaks. These findings suggest that decrotonylation and ubiquitination of H2A at lysine 119 act together to resolve replication stress-induced TRCs and protect genome stability.
Asunto(s)
Histonas , Lisina , ADN/metabolismo , Histonas/metabolismo , Lisina/metabolismo , Procesamiento Proteico-Postraduccional , ARN Polimerasa II/metabolismo , Sirtuina 1/genética , UbiquitinaciónRESUMEN
2205 duplex stainless steel (DSS) has good corrosion resistance due to its typical duplex organization, but the increasingly harsh CO2-containing oil and gas environment leads to different degrees of corrosion, especially pitting corrosion, which seriously threatens the safety and reliability of oil and gas development. In this paper, the effect of temperature on the corrosion behavior of 2205 DSS in a simulated solution containing 100 g/L Cl- and saturated CO2 was investigated with immersion tests and electrochemical tests and combined with characterization techniques such as laser confocal microscopy and X-ray photoelectron spectroscopy. The results show that the average critical pitting temperature of 2205 DSS was 66.9 °C. When the temperature was higher than 66.9 °C, the pitting breakdown potential, passivation interval, and self-corrosion potential decreased, while the dimensional passivation current density increased, and the pitting sensitivity was enhanced. With a further increase in temperature, the capacitive arc radius of 2205 DSS decreased, the film resistance and charge transfer resistance gradually decreased, the carrier density of the donor and acceptor in the product film layer with n + p bipolar characteristics also increased and the inner layer of the film with Cr oxide content decreased, while the outer layer with Fe oxide content increased, the dissolution of the film layer increased, the stability decreased, and the number and pore size of pits increased.
RESUMEN
In the study, seventeen angiotensin converting enzyme (ACE) inhibitory peptides were isolated from the protein hydrolysate of blue mussel (Mytilus edulis) and identified as MFR, MFV, FV, KP, QP, QVK, IK, YKV, IRK, MLKV, NFRPQ, YEGDP, WF, GPE, SWISS, SVEWK, and FKWH, respectively. Among them, IK, YEGDP, WF, and SWISS showed the strongest ACE inhibitory activity with IC50 values of 0.77 ± 0.020, 0.19 ± 0.010, 0.40 ± 0.015, and 0.32 ± 0.017 mg mL-1, respectively. Molecular docking study indicated that IK, YEGDP, WF, and SWISS exhibited better inhibitory activity attributed to its effective interaction with the active site of ACE by hydrogen bonding, electrostatic force and hydrophobic interaction. Furthermore, IK, YEGDP and WF perform an important protective function on human umbilical vein endothelial cells (HUVECs) by increasing nitric oxide (NO) content, decreasing endothelin-1 (ET-1) secretion, and antagonizing the adverse impact of norepinephrine on the secretion of NO and ET-1. In addition, YEGDP and WF could provide protection to HUVECs against H2O2 damage by increasing superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and NO levels to decrease the contents of reactive oxygen species (ROS) and malondialdehyde. Therefore, seventeen ACE inhibitory peptides, especially YEGDP and WF, might be used as natural ingredients for the development of products with antihypertensive functions.
Asunto(s)
Mytilus edulis , Hidrolisados de Proteína , Inhibidores de la Enzima Convertidora de Angiotensina/química , Animales , Células Endoteliales de la Vena Umbilical Humana , Humanos , Peróxido de Hidrógeno/farmacología , Simulación del Acoplamiento Molecular , Óxido Nítrico/metabolismo , Péptidos/química , Peptidil-Dipeptidasa A/química , Hidrolisados de Proteína/químicaRESUMEN
To effectively utilize skipjack tuna (Katsuwonus pelamis) processing by-products to prepare peptides with high angiotensin-I-converting enzyme (ACE) inhibitory (ACEi) activity, Neutrase was selected from five kinds of protease for hydrolyzing skipjack tuna dark muscle, and its best hydrolysis conditions were optimized as enzyme dose of 1.6%, pH 6.7, and temperature of 50°C using single factor and response surface experiments. Subsequently, 14 novel ACEi peptides were prepared from the high ACEi protein hydrolysate and identified as TE, AG, MWN, MEKS, VK, MQR, MKKS, VKRT, IPK, YNY, LPRS, FEK, IRR, and WERGE. MWN, MEKS, MKKS, and LPRS displayed significantly ACEi activity with IC50 values of 0.328 ± 0.035, 0.527 ± 0.030, 0.269 ± 0.006, and 0.495 ± 0.024 mg/mL, respectively. Furthermore, LPRS showed the highest increasing ability on nitric oxide (NO) production among four ACEi peptides combining the direct increase and reversing the negative influence of norepinephrine (NE), and MKKS showed the highest ability on directly decreasing and reversing the side effects of NE on the secretion level of endothelin-1 (ET-1) among four ACEi peptides. These findings demonstrate that seafood by-product proteins are potential ACEi peptide sources and prepared ACEi peptides from skipjack tuna dark muscle, which are beneficial components for functional food against hypertension and cardiovascular diseases.
RESUMEN
To prepare bioactive peptides with high angiotensin-I-converting enzyme (ACE)-inhibitory (ACEi) activity, Alcalase was selected from five kinds of protease for hydrolyzing Skipjack tuna (Katsuwonus pelamis) muscle, and its best hydrolysis conditions were optimized using single factor and response surface experiments. Then, the high ACEi protein hydrolysate (TMPH) of skipjack tuna muscle was prepared using Alcalase under the optimum conditions of enzyme dose 2.3%, enzymolysis temperature 56.2 °C, and pH 9.4, and its ACEi activity reached 72.71% at 1.0 mg/mL. Subsequently, six novel ACEi peptides were prepared from TMPH using ultrafiltration and chromatography methods and were identified as Ser-Pro (SP), Val-Asp-Arg-Tyr-Phe (VDRYF), Val-His-Gly-Val-Val (VHGVV), Tyr-Glu (YE), Phe-Glu-Met (FEM), and Phe-Trp-Arg-Val (FWRV), with molecular weights of 202.3, 698.9, 509.7, 310.4, 425.6, and 606.8 Da, respectively. SP and VDRYF displayed noticeable ACEi activity, with IC50 values of 0.06 ± 0.01 and 0.28 ± 0.03 mg/mL, respectively. Molecular docking analysis illustrated that the high ACEi activity of SP and VDRYF was attributed to effective interaction with the active sites/pockets of ACE by hydrogen bonding, electrostatic force, and hydrophobic interaction. Furthermore, SP and VDRYF could significantly up-regulate nitric oxide (NO) production and down-regulate endothelin-1 (ET-1) secretion in HUVECs after 24 h treatment, but also abolish the negative effect of 0.5 µM norepinephrine (NE) on the generation of NO and ET-1. Therefore, ACEi peptides derived from skipjack tuna (K. pelamis) muscle, especially SP and VDRYF, are beneficial components for functional food against hypertension and cardiovascular diseases.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina , Músculo Esquelético/química , Péptidos , Atún , Secuencia de Aminoácidos , Inhibidores de la Enzima Convertidora de Angiotensina/química , Inhibidores de la Enzima Convertidora de Angiotensina/aislamiento & purificación , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Endotelina-1/metabolismo , Alimentos Funcionales , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Hidrólisis , Simulación del Acoplamiento Molecular , Óxido Nítrico/metabolismo , Péptidos/química , Péptidos/aislamiento & purificación , Péptidos/farmacología , Hidrolisados de Proteína/química , Subtilisinas/químicaRESUMEN
Rice OsLIC encoding a CCCH zinc finger transcription factor plays an important role in immunity. However, the immune signaling pathways that OsLIC-involved and the underlying mechanisms that OsLIC-conferred resistance against pathogens are largely unclear. Here, we show that OsLIC, as a substrate for OsMAPK6, negatively regulates resistance to Xanthomonas oryzae pv. oryzae (Xoo) and X. oryzae pv. oryzicola (Xoc) by directly suppressing OsWRKY30 transcription. Biochemical assays showed that OsLIC bound to OsWRKY30 promoter and suppressed its transcription. Genetic assays confirmed that the osilc knockout mutants and OsWRKY30-overexpressing plants exhibited enhanced resistance to Xoo and Xoc, knocking out OsWRKY30 in the oslic mutants attenuated the resistance against bacterial pathogens. OsMAPK6 physically interacted with and phosphorylated OsLIC leading to decreased OsLIC DNA-binding activity, therefore, overexpression of OsLIC partially suppressed OsMAPK6-mediated rice resistance. In addition, both OsMAPK6-phosphorylated activation of OsLIC and phosphorylation-mimic OsLIC5D had reduced DNA-binding activity towards OsWRKY30 promoter, thereby promoting OsWRKY30 transcription. Collectively, these results reveal that OsMAPK6-mediated phosphorylation of OsLIC positively regulates rice resistance to Xoo and Xoc by modulating OsWRKY30 transcription, suggesting that OsMAPK6-OsLIC-OsWRKY30 module is an immune signaling pathway in response to the bacterial pathogens.
Asunto(s)
Oryza , Xanthomonas , ADN/metabolismo , Oryza/microbiología , Enfermedades de las Plantas/microbiología , Dedos de ZincRESUMEN
The objective of this study was to explore the application value of digital subtraction angiography (DSA) images optimized by deep learning algorithms in vascular restenosis patients undergoing cardiovascular intervention and their nursing efficacy. In this study, a network model for removing artifacts was constructed based on a deep algorithm. 60 patients with coronary artery restenosis were selected as the research objects, and they were randomly divided into the CTA group guided by CT angiography (CTA) and digital subtraction angiography (DSA) group, with 30 cases in each group. The antiartifact network model constructed based on the depth algorithm was applied to the images of CTA and DSA for experiments. After cardiovascular intervention and clinical pathway nursing intervention, it was found that the diameter stenosis rate in the DSA group decreased from 65.82 ± 12.9% to 4.7 ± 1.3%, and the area stenosis rate decreased from 88.4 ± 14.3% to 5.4 ± 1.7%. During the follow-up period of 3-24 months, 3 out of 46 lesions in the DSA group showed restenosis, so the restenosis rate was 6.5%, which was significantly lower than the 18.4% in the CTA group (P < 0.05). In the DSA group, there was 1 case of bleeding, 0 case of hematoma, 2 cases of urinary retention, and 0 case of hypotension, so the total incidence of adverse reactions was 10%, which was significantly lower than the 30% of the CTA group (P < 0.05). The high-sensitivity C-reactive protein (hs-CRP) levels of the two groups of patients were 3.58 ± 2.02 mg/L and 4.36 ± 3.11 mg/L before surgery and 3.49 ± 2.18 mg/L and 4.57 ± 3.4 mg/L after the surgery. The postoperative hs-CRP level in the CTA group was slightly lower than that before the surgery and the postoperative hs-CRP level in the DSA group was slightly higher than that before the surgery, but they were not statistically significant (P > 0.05). The hs-CRP level of the DSA group before and after the surgery was slightly higher than that of the CTA group, but there was no significant difference (P > 0.05). In summary, the network model based on the deep learning algorithm can remove the artifacts in DSA images and present high-quality clear images, and convolutional neural network (CNN) algorithms had a strong ability to automatically learn features in the field of medical image processing and were worthy of being widely used and popularized. In addition, the DSA-guided intervention can reduce the rate of vascular stenosis in patients, showing low probability of postoperative restenosis and adverse reactions and a good clinical effect.
Asunto(s)
Algoritmos , Angiografía de Substracción Digital/estadística & datos numéricos , Reestenosis Coronaria/diagnóstico por imagen , Aprendizaje Profundo , Adulto , Anciano , Artefactos , Proteína C-Reactiva/metabolismo , Biología Computacional , Angiografía por Tomografía Computarizada/estadística & datos numéricos , Reestenosis Coronaria/enfermería , Reestenosis Coronaria/terapia , Estenosis Coronaria/diagnóstico por imagen , Estenosis Coronaria/enfermería , Estenosis Coronaria/terapia , Femenino , Humanos , Masculino , Persona de Mediana Edad , Intervención Coronaria PercutáneaRESUMEN
The sea buckthorn (Hippophae rhamnoides L.) berries are rich in various bioactive components and widely used as fruit and traditional medicine. In this study, a novel heteropolysaccharide fraction (SP0.1-1) was isolated from Sea buckthorn berries. SP0.1-1 is composed of mannose, glucose, galactose, and arabinose in the molar ratio of 1:2.3:1.9:11.2 with a core structure containing 1,4-linked-α-d-Glcp, 1,4,6-linked-α-d-Glcp and 1,4-linked-α-d-Manp residues as the backbone. And the side-chains comprised of 1,3,5-linked-α-l-Araf, 1,5-linked-α-l-Araf, terminal α-Araf and 1,4-linked-ß-d-Galp. Furthermore, a diet supplemented with SP0.1-1 extended the mean lifespan, enhanced antioxidant enzyme (superoxide dismutase, SOD; glutathione peroxidase, GSH-Px; and catalase, CAT) activities, and decreased the malondialdehyde (MDA) level and hydrogen peroxide (H2O2)-induced mortality rate in fruit flies (Drosophila melanogaster). To summarize, the study's findings will provide evidence for the development of sea buckthorn polysaccharide products.
Asunto(s)
Envejecimiento/efectos de los fármacos , Antioxidantes , Drosophila melanogaster/efectos de los fármacos , Hippophae/metabolismo , Polisacáridos , Animales , Antioxidantes/química , Antioxidantes/farmacología , Polisacáridos/química , Polisacáridos/farmacologíaRESUMEN
miRNAs are important regulators of eukaryotic gene expression. The post-transcriptional maturation of miRNAs is controlled by the Drosha-DiGeorge syndrome critical region gene 8 (DGCR8) microprocessor. Dysregulation of miRNA biogenesis has been implicated in the pathogenesis of human diseases, including cancers. C-terminal-binding protein-interacting protein (CtIP) is a well-known DNA repair factor that promotes the processing of DNA double-strand break (DSB) to initiate homologous recombination-mediated DSB repair. However, it was unclear whether CtIP has other unknown cellular functions. Here, we aimed to uncover the roles of CtIP in miRNA maturation and cancer cell metastasis. We found that CtIP is a potential regulatory factor that suppresses the processing of miRNA primary transcripts (pri-miRNA). CtIP directly bound to both DGCR8 and pri-miRNAs through a conserved Sae2-like domain, reduced the binding of Drosha to DGCR8 and pri-miRNA substrate, and inhibited processing activity of Drosha complex. CtIP depletion significantly increased the expression levels of a subset of mature miRNAs, including miR-302 family members that are associated with tumor progression and metastasis in several cancer types. We also found that CtIP-inhibited miRNAs, such as miR-302 family members, are not crucial for DSB repair. However, increase of miR-302b levels or loss of CtIP function severely suppressed human colon cancer cell line tumor cell metastasis in a mouse xenograft model. These studies reveal a previously unrecognized mechanism of CtIP in miRNA processing and tumor metastasis that represents a new function of CtIP in cancer.
Asunto(s)
Transformación Celular Neoplásica , Neoplasias del Colon/patología , Endodesoxirribonucleasas/metabolismo , MicroARNs/genética , Animales , Línea Celular Tumoral , Humanos , Ratones , Metástasis de la Neoplasia , Proteínas Proto-Oncogénicas pp60(c-src)RESUMEN
A polysaccharide, CSL-0.1, was isolated from the medicinal lichen, Usnea longissima. CSL-0.1 was a neutral rhamnose-containing glucogalactomannan with a molecular weight of 7.86 × 104 Da. The polysaccharide had a core mannan structure with (1 â 6)-α-d-Manp units as the main chain and was substituted at the O-2 positions with side chains containing (1 â 2)-α-d-Manp residue, [3)-α-Glcp(1 â 4)-α-Glcp(1â] and 6-O-substituted ß-d-Galf units. 2-O- and 2,3-di-O-substituted Rhap units. The effects of CSL-0.1 on intestinal immunity and antioxidant activity were evaluated. CSL-0.1 increased the spleen and thymus indices in a dose-dependent manner and conferred immunomodulation on reversing the Th1/Th2-related cytokine imbalance in cyclophosphamide (CP)-induced immunosuppressed mice. CSL-0.1 could also enhance the levels of secretory immunoglobulin A in CP-injected mice. Additionally, the antioxidant levels in the liver and intestine of the mice were increased 20%-50% after intragastric injection by CSL-0.1.
Asunto(s)
Inmunización , Líquenes/química , Parmeliaceae/química , Polisacáridos/farmacología , Animales , Antioxidantes/farmacología , Espectroscopía de Resonancia Magnética con Carbono-13 , Citocinas/metabolismo , Femenino , Cromatografía de Gases y Espectrometría de Masas , Hidrólisis , Intestinos/efectos de los fármacos , Hígado/efectos de los fármacos , Hígado/metabolismo , Metilación , Ratones Endogámicos ICR , Monosacáridos/análisis , Polisacáridos/aislamiento & purificación , Espectroscopía de Protones por Resonancia Magnética , Espectrofotometría Infrarroja , Bazo/efectos de los fármacos , Timo/efectos de los fármacosRESUMEN
OBJECTIVE: To investigate the clinical efficacy of bronchial lavage under fiberoptic bronchoscopy in the treatment of severe pulmonary infection. METHODS: One hundred forty eight patients with severe pulmonary infection who were admitted to our hospital from October 2016 to December 2017 were included in this study. According to the random number table method, they were divided into a control group and an observation group with 79 patients each. The control group was given conventional treatment, while the observation group was given bronchoalveolar lavage with fiberoptic bronchoscopy on the basis of the treatment in the control group. The clinical efficacy of the two groups was compared, the duration of mechanical ventilation, antibiotic use and symptoms improvement of the two groups were recorded, and the respiratory mechanics parameters, serum procalcitonin (PCT) and transforming growth factor ß (TGF-ß) level were measured before and after treatment. RESULTS: The duration of mechanical ventilation, antibiotic use, respiratory failure correction, body temperature decline and white blood cell recovery in the observation group were significantly shorter than those in the control group (P<0.05). The total efficacy of the observation group was significantly higher than that of the control group (92.4% vs. 74.7%). The respiratory mechanics parameters of the two groups after treatment were higher than those before treatment (P<0.05) and the increase of the observation group was more obvious than that of the control group (P<0.05). The serum PCT and TGF-ß levels of the two groups after treatment were lower than those before treatment (P<0.05), and the decrease level in the observation group was more obvious (P<0.05). CONCLUSION: Bronchial lavage under fiberoptic bronchoscopy can improve the clinical efficacy, accelerate the improvement of clinical symptoms and respiratory mechanics parameters, significantly reduce the PCT and TGF-ß levels, and promote the rapid recovery of patients in the treatment of severe pulmonary infection.
RESUMEN
ETHNOPHARMACOLOGICAL RELEVANCE: Shiraia bambusicola is a parasitic fungus on the twigs of bamboos. Its relatively large stroma has high medicinal value and can treat a variety of diseases such as rheumatoid arthritis, cold stomach pain, sciatica, injuries, chronic bronchitis, and infantile. It is widely distributed in many provinces in Southern China and also is also found in Japan. AIM OF THE STUDY: Medicinal fungi were important resources for bioactive polysaccharides. To explore bioactive polysaccharides from Shiraia bambusicola, a heteropolysaccharide SB2-1 was purified and obtained from S. bambusicola and its immunostimulating activity was researched. MATERIALS AND METHODS: The polysaccharide from S. bambusicola was extracted and purified using enzyme assisted extraction, ethanol precipitation, anion-exchange and size-exclusion chromatography. Molecular weight of polysaccharide was estimated by high performance gel permeation chromatography. Monosaccharide compositions were determined by high performance liquid chromatography after pre-column derivatization and UV detection. Structure information was elucidated by IR spectrum, GC-MS analysis after methylation and gradual acid hydrolysis of the polysaccharide. The RAW264.7 cells were used to study the immunostimulating activity in vitro. RESULTS: Physicochemical and structural analyses showed that SB2-1 was a neutral heteropolysaccharide with molecular weight at 22.2 kDa and consisted of glucose, galactose and mannose at a ratio of 2.0:1.5:1.0. The structure of SB2-1 was a branched polysaccharides composed of a mannan core and side chains consisted of glucose and galactose. The mannan core was composed of (1â2)-Manp as the main chain. Glucose with (1â4)-D-Glcp, (1â2)-D-Glcp and (1â6)-D-Glcp at different degrees of polymerization were linked at C-6 and C-3 of the (1â2)-Manp as the side chains. The galactose with the linages of (1â6)-D-Galf, â2)-D-Galf(1â and terminal D-Galf(1â also existed in the side chain. The study on the immunostimulating activities of SB2-1 and its core structure P-2 were investigated on RAW264.7 macrophages. The results showed that SB2-1 could activate RAW264.7 macrophage and significantly improve its phagocytic ability by neutral red uptake experiment. Meanwhile, SB2-1 increased significantly higher inducible nitric oxide synthase (iNOS) production and the productions of IL-1, IL-6, IL-12 and TNF-α. The effect of SB2-1 was better than its core structure P-2 produced by gradual acid hydrolysis, which meant the side chains played an important role in the immunostimulating activities. CONCLUSIONS: The investigation demonstrated that the galactofuranose-containing mannogalactoglucan was characteristic polysaccharides in S. bambusicola and could enhance the activation of macrophages.
Asunto(s)
Adyuvantes Inmunológicos/química , Adyuvantes Inmunológicos/farmacología , Ascomicetos/química , Polisacáridos Fúngicos/química , Polisacáridos Fúngicos/farmacología , Animales , Supervivencia Celular/efectos de los fármacos , Ratones , Células RAW 264.7RESUMEN
BACKGROUND: We aimed to identify key genes and microRNAs (miRNAs) associated with the development of polycystic ovary syndrome (PCOS). METHODS: GSE84376 mRNA microarray data (15 PCOS granulosa cells and 13 control granulosa cells) and GSE34526 mRNA microarray data (7 PCOS granulosa cells and 3 control granulosa cells) were downloaded from the Gene Expression Omnibus (GEO) database. First, differentially expressed gene (DEG) analysis, gene set enrichment analysis (GSEA) for differentially expressed mRNAs, and protein-protein interaction (PPI) network analysis were conducted. Next, miRNA-target genes were analyzed and functions predicted, and a competing endogenous RNA (ceRNA) network was constructed. Finally, the relationship between miR-486-5p and PRELID2 was experimentally validated. RESULTS: Spleen tyrosine kinase (SYK), major histocompatibility complex, class II, DR alpha (HLA-DRA), and interleukin 10 (IL-10) were important nodes in the PPI network. Interestingly, HLA-DRA was significantly enriched in phagosomes mediated by Staphylococcus aureus infection, and in IL-10 enriched during S. aureus infection. One miRNA (miR-486-5p) and a single target gene (PRELID2) were obtained from the ceRNA network. Further experiments showed that miR-486-5p is upregulated and PRELID2 is downregulated in PCOS patient granulosa cells, and that miR-486-5p targets the PRELID2 3'UTR. Topological property analysis showed that hsa-miR-4687-5p downregulation and hsa-miR-4651 upregulation determined the levels of most mRNAs. Levels of the hsa-miR-4651 target gene were significantly enriched in the leukocyte transendothelial migration pathway. CONCLUSIONS: Our results suggest that HLA-DRA and IL-10 may contribute to PCOS progression via phagosome enriched by S. aureus infection, while miR-486-5p may be implicated in follicular development in PCOS by targeting PRELID2. Besides, miR-4651 may be involved in inflammation via leukocyte transendothelial migration, by regulating its target gene. These findings may indicate new directions and constitute a breakthrough in studying the pathophysiology of PCOS.
Asunto(s)
MicroARNs/genética , Síndrome del Ovario Poliquístico/genética , ARN Mensajero/genética , Adulto , Estudios de Casos y Controles , Células Cultivadas , Femenino , Perfilación de la Expresión Génica , Redes Reguladoras de Genes , Estudios de Asociación Genética , Predisposición Genética a la Enfermedad , Células de la Granulosa/metabolismo , Humanos , MicroARNs/metabolismo , Análisis por Micromatrices , Síndrome del Ovario Poliquístico/patología , ARN Mensajero/metabolismoRESUMEN
In the work, defatted muscle proteins of monkfish (Lophius litulon) were separately hydrolyzed by pepsin, trypsin, and in vitro gastrointestinal (GI) digestion methods, and antioxidant peptides were isolated from proteins hydrolysate of monkfish muscle using ultrafiltration and chromatography processes. The antioxidant activities of isolated peptides were evaluated using radical scavenging and lipid peroxidation assays and H2O2-induced model of HepG2 cells. In which, the cell viability, reactive oxygen species (ROS) content, and antioxidant enzymes and malondialdehyde (MDA) levels were measured for evaluating the protective extent on HepG2 cells damaged by H2O2. The results indicated that the hydrolysate (MPTH) prepared using in vitro GI digestion method showed the highest degree of hydrolysis (27.24 ± 1.57%) and scavenging activity on a 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical (44.54 ± 3.12%) and hydroxyl radical (41.32 ± 2.73%) at the concentration of 5 mg protein/mL among the three hydrolysates. Subsequently, thirteen antioxidant peptides (MMP-1 to MMP-13) were isolated from MPTH. According to their DPPH radical and hydroxyl radical scavenging activity, three peptides with the highest antioxidant activity were selected and identified as EDIVCW (MMP-4), MEPVW (MMP-7), and YWDAW (MMP-12) with molecular weights of 763.82, 660.75, and 739.75 Da, respectively. EDIVCW, MEPVW, and YWDAW showed high scavenging activities on DPPH radical (EC50 0.39, 0.62, and 0.51 mg/mL, respectively), hydroxyl radical (EC50 0.61, 0.38, and 0.32 mg/mL, respectively), and superoxide anion radical (EC50 0.76, 0.94, 0.48 mg/mL, respectively). EDIVCW and YWDAW showed equivalent inhibiting ability on lipid peroxidation with glutathione in the linoleic acid model system. Moreover, EDIVCW, MEPVW, and YWDAW had no cytotoxicity to HepG2 cells at the concentration of 100.0 µM and could concentration-dependently protect HepG2 cells from H2O2-induced oxidative damage through decreasing the levels of reactive oxygen species (ROS) and MDA and activating intracellular antioxidant enzymes of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px). These present results indicated that the protein hydrolysate and isolated antioxidant peptides from monkfish muscle, especially YWDAW could serve as powerful antioxidants applied in the treatment of some liver diseases and healthcare products associated with oxidative stress.