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Hemostasis is the first step of emergency medical treatment. It is particularly important to develop rapid-acting and efficacious hemostatic materials. Carboxymethyl chitosan (CMCS), sodium alginate (SA) and Resina Draconis (RD) were composited uniformly by polyelectrolyte blending. Their composite sponges (CMCS/SA/RD) were prepared by freeze-induced phase separation. CMCS/SA/RD sponges were characterized by Fourier transform infrared spectroscopy and scanning electron microscopy, and their blood absorption and hemolysis ratio were analyzed. The hemostatic effect of the composite sponges was evaluated by coagulation in vitro and in vivo. The composite sponges had a porous network structure. The water absorption ratio was >8000 %, and hemolysis ratio was <5 %. CMCS/SA/RD-II and CMCS/SA/RD-III composite sponges shortened the coagulation time in vitro by 11.33 s and 9.66 s, the hepatic hemostasis time by 13.8 % and 23.3 %, and the hemostasis time after mouse-tail amputation by 28.9 % and 23.9 %, respectively. A preliminary study on its coagulation mechanism showed that CMCS/SA/RD had significant effects on erythrocyte adsorption, platelet adhesion, and shortening of the activated partial thromboplastin time.
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Alginatos , Coagulación Sanguínea , Quitosano , Hemostasis , Hemostáticos , Quitosano/química , Quitosano/análogos & derivados , Alginatos/química , Animales , Hemostasis/efectos de los fármacos , Ratones , Hemostáticos/química , Hemostáticos/farmacología , Coagulación Sanguínea/efectos de los fármacos , Hemólisis/efectos de los fármacos , Adhesividad Plaquetaria/efectos de los fármacos , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
OBJECTIVE: To assess the effectiveness of a comprehensive rehabilitation approach combining Traditional Chinese Medicine Daoyin with lower limb robotics during the recovery phase of stroke patients. METHODS: Stroke patients meeting the specified criteria were randomly assigned to one of four groups using a random number table: Control group, Daoyin group, lower limb robot group (LLR group), and Daoyin and lower limb robot group (DLLR group). Each group received distinct treatments based on conventional rehabilitation training. The treatment duration spanned two weeks with two days of rest per week. Pre- and post-intervention assessments included various scales: Fugl-Meyer Assessment (FMA), Berg balance scale (BBS), Barthel index (BI), Fatigue Scale-14 (FS-14), Pittsburgh sleep quality index (PSQI), Hamilton Anxiety Scale (HAMA), and Hamilton Depression Scale (HAMD). RESULTS: Statistically significant differences were observed in the lower limb function measured by FAM between the Control group (15 ± 5) and the DLLR group (18 ± 5) (P = 0.049). In the Barthel index, a statistically significant difference was noted between the Control group (54 ± 18) and the DLLR group (64 ± 11) (P = 0.041). Additionally, significant differences were found in the Berg balance scale between the Control group (21 ± 10) and the DLLR group (27 ± 8) (P = 0.024), as well as between the Control group (21 ± 10) and the LLR group (26 ± 10) (P = 0.048). CONCLUSION: The findings of this study suggest that the combined use of Daoyin and robotics not only enhances motor function in stroke patients but also has a positive impact on fatigue, sleep quality, and mood. This approach may offer a more effective rehabilitation strategy for stroke patients.
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Medicamentos Herbarios Chinos , Extremidad Inferior , Robótica , Rehabilitación de Accidente Cerebrovascular , Accidente Cerebrovascular , Humanos , Masculino , Persona de Mediana Edad , Femenino , Robótica/instrumentación , Anciano , Extremidad Inferior/fisiopatología , Accidente Cerebrovascular/fisiopatología , Rehabilitación de Accidente Cerebrovascular/métodos , Medicamentos Herbarios Chinos/uso terapéutico , Resultado del Tratamiento , AdultoRESUMEN
Biomolecules play vital roles in many biological processes and diseases, making their identification crucial. Herein, we present a colorimetric sensing method for detecting biomolecules like cysteine (Cys), homocysteine (Hcy), and glutathione (GSH). This approach is based on a reaction system whereby colorless 3,3',5,5'-tetramethylbenzidine (TMB) undergoes catalytic oxidation to form blue-colored oxidized TMB (ox-TMB) in the presence of hydrogen peroxide (H2O2), utilizing the peroxidase and catalase-mimicking activities of metal-phenolic coordination frameworks (MPNs) of Cu-TA, Co-TA, and Fe-TA nanospheres. The Fe-TA nanospheres demonstrated superior activity, more active sites and enhanced electron transport. Under optimal conditions, the Fe-TA nanospheres were used for the detection of biomolecules. When present, biomolecules inhibit the reaction between TMB and H2O2, causing various colorimetric responses at low detection limits of 0.382, 0.776 and 0.750 µM for Cys, Hcy and GSH. Furthermore, it was successfully applied to real water samples with good recovery results. The developed sensor not only offers a rapid, portable, and user-friendly technique for multi-target analysis of biomolecules at low concentrations but also expands the potential uses of MPNs for other targets in the environmental field.
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Bencidinas , Colorimetría , Cisteína , Glutatión , Peróxido de Hidrógeno , Colorimetría/métodos , Peróxido de Hidrógeno/química , Glutatión/química , Glutatión/análisis , Cisteína/química , Cisteína/análisis , Bencidinas/química , Homocisteína/análisis , Homocisteína/química , Estructuras Metalorgánicas/química , Límite de Detección , Fenoles/química , Fenoles/análisis , Oxidación-Reducción , Catálisis , Peroxidasa/química , Catalasa/químicaRESUMEN
White matter injury (WMI) causes oligodendrocyte precursor cell (OPC) differentiation arrest and functional deficits, with no effective therapies to date. Here, we report increased expression of growth hormone (GH) in the hypoxic neonatal mouse brain, a model of WMI. GH treatment during or post hypoxic exposure rescues hypoxia-induced hypomyelination and promotes functional recovery in adolescent mice. Single-cell sequencing reveals that Ghr mRNA expression is highly enriched in vascular cells. Cell-lineage labeling and tracing identify the GHR-expressing vascular cells as a subpopulation of pericytes. These cells display tip-cell-like morphology with kinetic polarized filopodia revealed by two-photon live imaging and seemingly direct blood vessel branching and bridging. Gain-of-function and loss-of-function experiments indicate that GHR signaling in pericytes is sufficient to modulate angiogenesis in neonatal brains, which enhances OPC differentiation and myelination indirectly. These findings demonstrate that targeting GHR and/or downstream effectors may represent a promising therapeutic strategy for WMI.
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Vaina de Mielina , Neovascularización Fisiológica , Pericitos , Animales , Pericitos/metabolismo , Pericitos/efectos de los fármacos , Ratones , Vaina de Mielina/metabolismo , Neovascularización Fisiológica/efectos de los fármacos , Neovascularización Fisiológica/fisiología , Hormona del Crecimiento/metabolismo , Hormona del Crecimiento/farmacología , Animales Recién Nacidos , Hipoxia/metabolismo , Diferenciación Celular/efectos de los fármacos , Ratones Endogámicos C57BL , Células Precursoras de Oligodendrocitos/metabolismo , Células Precursoras de Oligodendrocitos/efectos de los fármacos , Receptores de Somatotropina/metabolismo , Receptores de Somatotropina/genética , AngiogénesisRESUMEN
A facile and efficient radical tandem vinylogous aldol and intramolecular [2 + 2] cycloaddition reaction for direct synthesis of cyclobutane-containing benzocyclobutenes (BCBs) under extremely mild conditions without using any photocatalysts is reported. This approach exhibited definite compatibility with functional groups and afforded new BCBs with excellent regioselectivity and high yields. Moreover, detailed mechanism studies were carried out both experimentally and theoretically. The readily accessible, low-cost, and ecofriendly nature of the developed strategy will endow it with attractive applications in organic and medicinal chemistry.
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Poly(amidoxime) (PAO) has been recognized as the most potential candidate for extracting uranium from seawater, owing to its merits of outstanding uranium affinity, low cost, and large-scale production. Despite remarkable achievements, existing PAO sorbents suffer from unsatisfactory uranium extraction efficiency and selectivity, as imposed by the inherently sluggish uranium adsorption kinetics and inevitable spatial configuration transition of amidoxime, which diminishes uranium affinity. Herein, we discover a facile and integrated design to elaborate a PAO/MXene nanocomposite that delivers ultrahigh and durable uranium/vanadium (U/V) selectivity. The key to our design lies in harnessing MXene-enabled strong intermolecular interactions to PAO to minimize the spatial configuration transition of amidoxime and stabilizing its superior uranium affinity, as well as creating a separated photothermal interface to maximize temperature-strengthened affinity for uranium over vanadium. Such a synergetic effect allows the nanocomposite to acquire over a 4-fold improvement in U/V selectivity compared to that of pure PAO as well as an unprecedented distribution coefficient of uranium compared to most state-of-the-art sorbents. We further demonstrate that our nanocomposite exhibits durable U/V selectivity with negligible attenuation and good antibacterial ability even in long-term operation. The design concept and extraordinary performance in this study bring PAO-based sorbents a step closer to practical uranium extraction from seawater.
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Bacterial infections have always been a major threat to public health, and the development of effective antibacterial substances from natural polymers is crucial. 2-Aminoisonicotinic acid (AN) was grafted onto chitosan by 1-ethyl-(3-dimethylaminopropyl)carbodiimide-mediated coupling reactions, and then modified chitosaniodine (CSAN-I) complexes were prepared by solvent-assisted grinding. The samples were characterized using ultraviolet-visible spectroscopy, Fourier transform infrared spectroscopy, Raman spectroscopy, proton nuclear magnetic resonance spectroscopy, and X-ray diffraction, confirming that CSAN-I complexes had been successfully prepared. Thermogravimetric (TG) analysis indicated that the chemical modification of chitosan and iodine complexation reduced the thermal stability; X-ray photoelectron spectroscopy (XPS) analysis revealed that 81 % of the iodine in CSAN-I complex was in the form of triiodide ions. The iodine contents of three CSAN-I complexes (CSAN-I-1, CSAN-I-2 and CSAN-I-3) were 1.59 ± 0.22 %, 3.18 ± 0.26 %, and 5.56 ± 0.41 %, respectively. The antibacterial effects were evaluated in vitro, and the results indicated that CSAN-I complexes had strong antibacterial activities against both E. coli and S. aureus. In particular, CSAN-I-3 exhibited the best antibacterial effect. In addition, CSAN-I-3 was nontoxic to L929 cells with good cytocompatibility. Therefore, CSAN-I complexes can be considered as promising candidates for wound management in clinical applications.
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Quitosano , Yodo , Quitosano/farmacología , Quitosano/química , Escherichia coli , Staphylococcus aureus , Yodo/farmacología , Antibacterianos/farmacología , Antibacterianos/química , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos X , Pruebas de Sensibilidad MicrobianaRESUMEN
Reactive oxygen species (ROS) act as a group of signaling molecules in rice functioning in regulation of development and stress responses. Respiratory burst oxidase homologues (Rbohs) are key enzymes in generation of ROS. However, the role of the nine Rboh family members was not fully understood in rice multiple disease resistance and yield traits. In this study, we constructed mutants of each Rboh genes and detected their requirement in rice multiple disease resistance and yield traits. Our results revealed that mutations of five Rboh genes (RbohA, RbohB, RbohE, RbohH, and RbohI) lead to compromised rice blast disease resistance in a disease nursery and lab conditions; mutations of five Rbohs (RbohA, RbohB, RbohC, RbohE, and RbohH) result in suppressed rice sheath blight resistance in a disease nursery and lab conditions; mutations of six Rbohs (RbohA, RbohB, RbohC, RbohE, RbohH and RbohI) lead to decreased rice leaf blight resistance in a paddy yard and ROS production induced by PAMPs and pathogen. Moreover, all Rboh genes participate in the regulation of rice yield traits, for all rboh mutants display one or more compromised yield traits, such as panicle number, grain number per panicle, seed setting rate, and grain weight, resulting in reduced yield per plant except rbohb and rbohf. Our results identified the Rboh family members involved in the regulation of rice resistance against multiple pathogens that caused the most serious diseases worldwide and provide theoretical supporting for breeding application of these Rbohs to coordinate rice disease resistance and yield traits.
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Peritoneal fibrosis is a severe clinical complication associated with peritoneal dialysis (PD) and impacts its efficacy and patient outcomes. The process of mesothelial-mesenchymal transition (MMT) in peritoneal mesothelial cells plays a pivotal role in fibrogenesis, whereas metabolic reprogramming, characterized by excessive glycolysis, is essential in MMT development. No reliable therapies are available despite substantial progress made in understanding the mechanisms underlying peritoneal fibrosis. Protective effect of omega-3 polyunsaturated fatty acids (ω3 PUFAs) has been described in PD-induced peritoneal fibrosis, although the detailed mechanisms remain unknown. It is known that ω3 PUFAs bind to and activate the free fatty acid receptor 4 (FFAR4). However, the expression and role of FFAR4 in the peritoneum have not been investigated. Thus, we hypothesized that ω3 PUFAs would alleviate peritoneal fibrosis by inhibiting hyperglycolysis and MMT through FFAR4 activation. First, we determined FFAR4 expression in peritoneal mesothelium in humans and mice. FFAR4 expression was abnormally decreased in patients on PD and mice and HMrSV5 mesothelial cells exposed to PD fluid (PDF); this change was restored by the ω3 PUFAs (EPA and DHA). ω3 PUFAs significantly inhibited peritoneal hyperglycolysis, MMT, and fibrosis in PDF-treated mice and HMrSV5 mesothelial cells; these changes induced by ω3 PUFAs were blunted by treatment with the FFAR4 antagonist AH7614 and FFAR4 siRNA. Additionally, ω3 PUFAs induced FFAR4, Ca2+/calmodulin-dependent protein kinase kinase ß (CaMKKß), and AMPK and suppressed mTOR, leading to the inhibition of hyperglycolysis, demonstrating that the ω3 PUFAs-mediated FFAR4 activation ameliorated peritoneal fibrosis by inhibiting hyperglycolysis and MMT via CaMKKß/AMPK/mTOR signaling. As natural FFAR4 agonists, ω3 PUFAs may be considered for the treatment of PD-associated peritoneal fibrosis.
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Ácidos Grasos Omega-3 , Fibrosis Peritoneal , Humanos , Ratones , Animales , Fibrosis Peritoneal/metabolismo , Quinasa de la Proteína Quinasa Dependiente de Calcio-Calmodulina/metabolismo , Proteínas Quinasas Activadas por AMP/metabolismo , Transducción de Señal , Serina-Treonina Quinasas TOR/metabolismoRESUMEN
Arabidopsis RESISTANCE TO POWDERY MILDEW 8.1 (RPW8.1) is an important tool for engineering broad-spectrum disease resistance against multiple pathogens. Ectopic expression of RPW8.1 leads to enhanced disease resistance with cell death at leaves and compromised plant growth, implying a regulatory mechanism balancing RPW8.1-mediated resistance and growth. Here, we show that RPW8.1 constitutively enhances the expression of transcription factor WRKY51 and activates salicylic acid and ethylene signalling pathways; WRKY51 in turn suppresses RPW8.1 expression, forming a feedback regulation loop. RPW8.1 and WRKY51 are both induced by pathogen infection and pathogen-/microbe-associated molecular patterns. In ectopic expression of RPW8.1 background (R1Y4), overexpression of WRKY51 not only rescues the growth suppression and cell death caused by RPW8.1, but also suppresses RPW8.1-mediated broad-spectrum disease resistance and pattern-triggered immunity. Mechanistically, WRKY51 directly binds to and represses RPW8.1 promoter, thus limiting the expression amplitude of RPW8.1. Moreover, WRKY6, WRKY28 and WRKY41 play a role redundant to WRKY51 in the suppression of RPW8.1 expression and are constitutively upregulated in R1Y4 plants with WRKY51 being knocked out (wrky51 R1Y4) plants. Notably, WRKY51 has no significant effects on disease resistance or plant growth in wild type without RPW8.1, indicating a specific role in RPW8.1-mediated disease resistance. Altogether, our results reveal a regulatory circuit controlling the accumulation of RPW8.1 to an appropriate level to precisely balance growth and disease resistance during pathogen invasion.
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Proteínas de Arabidopsis , Arabidopsis , Proteínas de Arabidopsis/metabolismo , Resistencia a la Enfermedad/genética , Retroalimentación , Arabidopsis/metabolismo , Muerte Celular , Enfermedades de las Plantas/genética , Regulación de la Expresión Génica de las Plantas/genéticaRESUMEN
Objective To observe the effects of Xixin Decoction on the blood-brain barrier permeability and the expressions of P-glycoprotein(P-gp),cannabinoid receptor 1(CB1)and cannabinoid receptor 2(CB2)in hippocampal tissue of rapidly aging mice(SAMP8);To explore the possible mechanism of Xixin Decoction in the treatment of Alzheimer disease(AD).Methods Totally 60 SAMP8 mice were randomly divided into model group,probiotics group,and Xixin Decoction high-,medium-and low-dosage groups,with 12 mice in each group,another 12 SAMR1 mice were set as control group.The medicated groups received corresponding drugs by gavage for 10 weeks respectively,while the control group and model group were gavaged with equal volume of distilled water.Morris water maze test was used to detect the learning and memory ability of mice,the blood-brain barrier permeability was detected by Evans blue method,the contents of matrix metalloproteinase 9(MMP9),nuclear factor(NF)-κB and tumor necrosis factor-α(TNF-α)in serum were determined by ELISA,the expressions of P-gp,CB1 and CB2 in hippocampal tissue were detected by Western blot,P-gp expression in hippocampal tissue was detected by immunofluorescence staining.Results Compared with the control group,the learning and memory ability of mice in model group significantly decreased,Evans blue exudation in brain tissue significantly increased,the contents of MMP9,TNF-α and NF-κB in serum significantly increased,the expressions of P-gp and CB2 protein significantly decreased,the expression of CB1 protein significantly increased,with statistical significance(P<0.01,P<0.05).Compared with the model group,the learning and memory ability of mice in Xixin Decoction high-dosage group significantly increased,the Evans blue exudation in brain tissue significantly decreased,the contents of MMP9,TNF-α and NF-κB in serum significantly decreased,the protein expressions of P-gp and CB2 significantly increased,and the protein expression of CB1 significantly decreased,with statistical significance(P<0.01,P<0.05).Conclusion Xixin Decoction can improve the spatial learning and memory ability of AD model mice,and its mechanism is related to regulating the permeability of the blood-brain barrier and related protein expression,and inhibiting neuroinflammation.
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Lower limb ischemia-reperfusion is a common pathological process during clinical surgery. Because lower limb ischemia-reperfusion usually aggravates ischemia-induced skeletal muscle tissue injury after lower limb ischemia-reperfusion, it also causes remote organ heart, intestine, liver, lung and other injuries, and there is no effective clinical treatment for lower limb ischemia-reperfusion injury, so it is urgent to study its injury mechanism. In this study, the rat model of lower limb ischemia-reperfusion was established by clamping the femoral artery with microarterial clips, and the wall destruction such as intimal injury, cell edema, collagen degeneration, neutrophil infiltration, and elastic fiberboard injury of the femoral artery wall was detected. The expression of inflammatory factors was detected by immunohistochemistry. miR-206 preconditioning was used to observe the expression of inflammatory factors, redox status and apoptosis in the vascular wall of rats after acute limb ischemia-reperfusion. Our findings suggest that vascular endothelial cell edema increases, wall thickening, neutrophil infiltration, and elastic fiber layer damage during IRI. Inflammatory factor expression was increased in femoral artery tissue, and miR-206 expression levels were significantly down-regulated. Further studies have found that miR-206 attenuates lower limb IRI by regulating the effects of phase inflammatory factors. In this study, we investigated the effect of miR-206 on inflammatory factors and its possible role in the development of lower limb IRI, providing new research ideas for the regulatory mechanism of lower limb IRI, and providing a certain theoretical basis for the treatment of lower limb ischemia-reperfusion injury after surgery or endovascular intervention.
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MicroARNs , Daño por Reperfusión , Ratas , Animales , Isquemia , Daño por Reperfusión/metabolismo , Extremidad Inferior/patología , MicroARNs/genética , MicroARNs/uso terapéutico , Edema , Modelos Animales de EnfermedadRESUMEN
Infectious diseases pose a major challenge to human health, and there is an urgent need to develop new antimicrobial agents with excellent antibacterial activity. A series of novel triazolo[4,3-a]pyrazine derivatives were synthesized and their structures were characterized using various techniques, such as melting point, 1H and 13C nuclear magnetic resonance spectroscopy, mass spectrometry, and elemental analysis. All the synthesized compounds were evaluated for in vitro antibacterial activity using the microbroth dilution method. Among all the tested compounds, some showed moderate to good antibacterial activities against both Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli strains. In particular, compound 2e exhibited superior antibacterial activities (MICs: 32 µg/mL against Staphylococcus aureus and 16 µg/mL against Escherichia coli), which was comparable to the first-line antibacterial agent ampicillin. In addition, the structure-activity relationship of the triazolo[4,3-a]pyrazine derivatives was preliminarily investigated.
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Antiinfecciosos , Infecciones Estafilocócicas , Humanos , Pirazinas/farmacología , Antibacterianos/química , Escherichia coli , Relación Estructura-Actividad , Pruebas de Sensibilidad Microbiana , Estructura MolecularRESUMEN
The widespread prevalence of infectious bacteria is one of the greatest threats to public health, and consequently, there is an urgent need for efficient and broad-spectrum antibacterial materials that are antibiotic-free. In this study, 2-pyridinecarboxaldehyde (PCA) was grafted onto chitosan (CS) and the modified CS coordinated with silver ions to prepare PCA-CS-Ag complexes with antibacterial activity. To obtain complexes with a high silver content, the preparation process was optimized using single-factor experiments and response surface methodology. Under the optimal preparation conditions (an additional amount of silver nitrate (58 mg), a solution pH of 3.9, and a reaction temperature of 69 °C), the silver content of the PCA-CS-Ag complex reached 13.27 mg/g. The structure of the PCA-CS-Ag complex was subsequently verified using ultraviolet-visible spectroscopy, Fourier-transform infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, and thermogravimetric analysis. Furthermore, three possible complexation modes of the PCA-CS-Ag complex were proposed using molecular mechanics calculations. The results of the antibacterial assay in vitro showed that the PCA-CS-Ag complex exhibited strong antibacterial activity against both Gram-positive and Gram-negative bacteria, exerting the synergistic antibacterial effect of modified chitosan and silver ions. Therefore, the PCA-CS-Ag complex is expected to be developed as an effective antibacterial material with promising applications in food films, packaging, medical dressings, and other fields.
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Quitosano , Nanopartículas del Metal , Antibacterianos/farmacología , Antibacterianos/química , Quitosano/química , Bacterias Gramnegativas , Bacterias Grampositivas , Espectroscopía Infrarroja por Transformada de Fourier , Iones , Nanopartículas del Metal/química , Pruebas de Sensibilidad MicrobianaRESUMEN
Ginsenosides are major bioactive compounds found in Panax ginseng that exhibit various pharmaceutical properties. Dammarenediol-II, the nucleus of dammarane-type ginsenosides, is a promising candidate for pharmacologically active triterpenes. Dammarenediol-II synthase (DDS) cyclizes 2,3-oxidosqualene to produce dammarenediol-II. Based on the native terpenoids synthetic pathway, a dammarane-type ginsenosides synthetic pathway was established in Chlamydomonas reinhardtii by introducing P. ginseng PgDDS, CYP450 enzyme (PgCYP716A47), or/and Arabidopsis thaliana NADPH-cytochrome P450 reductase gene (AtCPR), which is responsible for producing dammarane-type ginsenosides. To enhance productivity, strategies such as "gene loading" and "culture optimizing" were employed. Multiple copies of transgene expression cassettes were introduced into the genome to increase the expression of the key rate-limiting enzyme gene, PgDDS, significantly improving the titer of dammarenediol-II to approximately 0.2 mg/L. Following the culture optimization in an opt2 medium supplemented with 1.5 mM methyl jasmonate under a light:dark regimen, the titer of dammarenediol-II increased more than 13-fold to approximately 2.6 mg/L. The C. reinhardtii strains engineered in this study constitute a good platform for the further production of ginsenosides in microalgae.
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Chlamydomonas reinhardtii , Ginsenósidos , Panax , Triterpenos , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Triterpenos/metabolismo , Panax/genética , DamaranosRESUMEN
Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) is a key regulator of sperm function and physiological metabolism. Metformin, an inexpensive and effective antioxidant, is known to play an important role in the activation of AMPK. Therefore metformin has potential to improve sperm cryopreservation. The aim of this study was to investigate the effect of metformin during semen cryopreservation of sheep and to find the most effective concentration in freezing extender. Semen were cryopreserved with extender containing different concentrations of metformin (0, 0.25, 0.5, 1.0, 2.0 and 4.0 mmol/L). Sperm motility, acrosome integrity and plasma membrane integrity were measured after semen freezing and thawing. All results showed that sperm quality was significantly increased in the 1.0 mmol/L metformin-treated group compared with the control group (P < 0.05). In addition, the study showed that metformin effectively reduced the content of malondialdehyde (MDA) and reactive oxygen species (ROS), and increased the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) of freeze-thawed sperm (P < 0.05). The optimal concentration of metformin was 1.0 mmol/L. Moreover, the results showed that AMPK was localized in the acrosome region, junction and midsection of sperm, and p-AMPK was distributed in the post-acrosomal region, junction and midsection. Western blot analysis indicated that 1.0 mmol/L metformin stimulated the phosphorylation of AMPK in sperm. Further results showed that 1.0 mmol/L metformin significantly increased the mitochondrial membrane potential (ΔΨm), ATP content, glucose uptake and lactate efflux of post-thawed sperm through the AMPK pathway, improved sperm quality, and increased the cleavage rate of in vitro fertilization (P < 0.05).
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Proteínas Quinasas Activadas por AMP , Semen , Masculino , Animales , Ovinos , Antioxidantes/farmacología , Motilidad Espermática , Criopreservación/veterinaria , EspermatozoidesRESUMEN
The microalgae Haematococcus pluvialis is a commercial source of natural astaxanthin. However, mature cells develop rigid three-layer wall structures and a repulsive odor. This study applied a liquid static fermentation system to screen hydrolyzing microorganisms for cell wall hydrolysis. Baijiu jiuqu and Gutian hongqu were found to have promising potential for application. The fermentation using 2% baijiu jiuqu and 2% glucose for pre-activation achieved comparable recovery of carotenoids to homogenizer disruption methods and produced stable fragrance which may be attributed to ethyl octanoate, hexyl formate, and phenethyl butyrate, as revealed by gas chromatography-mass spectrometry analysis. The abundance of astaxanthin molecules was slightly affected by fermentation with fold change < 2, while molecules with higher fold change (>10) were mainly carbohydrates, lipids, and steroids proving the safety of the fermentation. This study provides a new scheme for the biorefining of Haematococcus. pluvialis, potentially contributing to the industrial production of natural astaxanthin.
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Chlorophyceae , Fermentación , Biomasa , Bebidas , Pared CelularRESUMEN
Considering the great harm to the human body caused by severe and massive bleeding, in this study, chitosan-grafted norfloxacin (CTS-NF) composites were prepared with chitosan (CTS) and norfloxacin (NF) as raw materials by a 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide-mediated coupling method to solve the limitations of slow hemostatic and poor anti-infective effects of current dressings on the market. The effects of the mass ratio of CTS to NF (MCTS/MNF), reaction temperature T and reaction time t on the grafting rate (η%) of the products were investigated through single factor tests. The preparation process was optimized with the η% as an evaluation index by means of the Box-Behnken test design and response surface analysis. The antimicrobial activity was evaluated by inhibition zone assay, and the hemostatic activity of the prepared composites was evaluated in vitro and in vivo. The results suggested that the optimum preparation conditions were the mass ratio of CTS to NF (MCTS/MNF) 5:3, reaction temperature 65 °C, and reaction time 4 h. Under this condition, the η% of CTS-NF was 45.5%. The CTS-NF composites displayed significant antimicrobial activities. Moreover, in vitro hemostasis results revealed that the CTS-NF composite had a lower blood clotting index and absorbed red blood cells to promote aggregation. In vivo ear and live hemostasis, the CTS-NF groups showed short hemostatic time (49.75 ± 3.32 s and 50.00 ± 7.21 s) and more blood loss (0.07 ± 0.010 g and 0.075 ± 0.013 g). The results showed that CTS-NF reduced the bleeding time and volume, exhibiting a significant coagulation effect. Therefore, the CTS-NF sponge is expected to be a new, effective hemostatic and antibacterial material in the future.
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OBJECTIVE To investigate the efficacy and safety of aerosol inhalation of polycolistin B in the treatment of severe pneumonia combined with mechanical ventilation, and to provide a reference of real-world data for clinical drug therapy. METHODS A retrospective cohort study was conducted to analyze the clinical data of 87 patients with severe pneumonia combined with mechanical ventilation at the First Affiliated Hospital of Shandong First Medical University from January 2021 to February 2023. According to route of administration, all patients were divided into combined group (24 cases, receiving aerosol YXH2021ZX013) inhibition of polycolistin B combined with intravenous dripping) and routine group (63 cases, intravenous dripping of polycolistin B alone). The differences in efficacy (mortality,clinical response rate and bacterial clearance rate)and safety (elevated serum creatinine, bronchospasm and skin pigmentation) were compared between two groups; the influential factors of primary outcome index as mortality were analyzed through univariate analysis and multivariate Logistic regression analysis. RESULTS In terms of efficacy, there were no statistical differences in mortality(37.50% vs. 41.27%, P=0.749), clinical response rate (54.17% vs. 55.56%, P=0.907) and bacterial clearance rate (45.83% vs. 44.44%, P=0.907) between the two groups. In terms of safety, the incidence of bronchospasm in the combined group was significantly higher than that of the routine group (12.50% vs. 0, P=0.028), but the differences in incidence of elevated serum creatinine and skin pigmentation between two groups were not statistically significant (P>0.05). Univariate analysis showed that the moralities were higher in the case of infected with Acinetobacter baumannii, Klebsiella pneumoniae and combined use of quinolones (P<0.05); multivariate Logistic regression analysis showed that infection with A. baumannii (OR=3.36, P=0.014) and combined use of quinolones (OR=3.54, P=0.013) were independently associated with mortality (P<0.05). CONCLUSIONS For severe pneumonia patients with mechanical ventilation, intravenous dripping of polycolistin B combined with aerosol inhalation does not show superior efficacy compared with intravenous dipping of polycolistin B alone, but significantly increases the incidence of bronchospasm. Infection with A. baumannii and combined use of quinolones are independent risk factors for the increase of mortality.
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ObjectiveTo observe the possible mechanism of Xixin Decoction (洗心汤, XXD) in the prevention and treatment of Alzheimer 's disease(AD). MethodsFifty rapid aging model mice (SAMP8) were randomly divided into model group, probiotic group, high-, moderate- and low-dose group of XXD, with 10 mice in each group. Another 10 homologous anti-rapid aging mice (SAMR1) were set as control group. After 10 weeks of feeding, the control group and the model group were given 10 ml·kg-1·d-1 of distilled water by gavage, while the probiotic group (0.39 g·kg-1·d-1), the high-dose group of XXD (5.08 g·kg-1·d-1), the moderate-dose group of XXD (2.54 g·kg-1·d-1), and the low-dose group of XXD (1.27 g·kg-1·d-1) were given corresponding drugs or decoctions by gavage, once a day in all groups. After 10 weeks of intragastric administration, Morris water maze was used to detect the spatial learning and memory ability of mice in each group. HE staining was used to observe the pathological changes of hippocampal CA3 region and colon. The levels of β-amyloid 1-42 (Aβ1-42), lipopolysaccharide (LPS), serum amyloid A (SAA) and acetylcholine (ACH) in hippocampus and colon were detected by ELISA.The diversity of intestinal flora in mouse feces was detected by 16S rRNA sequencing. ResultsCompared to those in the control group, the levels of Aβ1-42,LPS, SAA increased, while the level of ACH decreased in the model group (P<0.05 or P<0.01). Compared to those in the model group, the escape latency period of the probiotic group was significantly shortened on the 2nd and 5th days, while the escape latency period was shortened, and the residence time in the target platform quadrant increased in the high-dose XXD group during the 2nd to 5th days; the escape latency period was shortened significantly in the moderate-dose XXD group on the 5th day (P<0.05 or P<0.01). Compared to those in the model group, the hippocampal neuron cells in the high- and moderate-dose XXD groups were arranged more closely, with decreased levels of SAA, Aβ1-42 and LPS, increased ACH level, Simpson and Shannon index (P<0.05 or P<0.01); the arrangement of hippocampal neuron cells in the probiotic group and the low-dose XXD group was relatively loose; the proportions of Bacteroidetes and Prevotella were significantly reduced in the probiotic group and the high-dose XXD group, while that of Firmicutes and Lactobacillus significantly increased (P<0.05 or P<0.01). Compared to those in the probiotic group and the high-dose XXD group, the number of goblet cells in the moderate-dose XXD group decreased, and the number of glands in the low-dose XXD group decreased with atrophy. The high-dose XXD group had decreased Aβ1-42 level in the hippocampus, increased ACH level in thehippocampus and colon tissue, and decreased SAA in the colon tissue than the moderate- and low-dose XXD groups (P<0.05 or P<0.01); moreover, the SAA level in the hippocampus was significantly higher in the low-dose XXD group than the high- and moderate-dose groups (P<0.01). ConclusionXXD can improve the spatial learning and memory ability of SAMP8, reduce the production and deposition of LPS, SAA and Aβ1-42 in brain and intestine, and increase the content of ACH. The mechanism of its prevention and treatment of AD maybe related to regulating intestinal microecology, affecting flora diversity and improving inflammatory response.