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Manufacturing is undergoing profound transformations, among which green biomanufacturing with low energy consumption, high efficiency, and sustainability is becoming one of the major trends. However, enzymes, as the "core chip" of biomanufacturing, are often handicapped in their application by their high cost, low operational stability, and nonreusability. Immobilization of enzymes is a technology that binds or restricts enzymes in a certain area with solid materials, allows them to still carry out their unique catalytic reaction, and allows them to be recycled and reused. Compared with free enzymes, immobilized enzymes boast numerous advantages such as enhanced storage stability, ease of separation, reusability, and controlled operation. Currently, commonly used supports for enzyme immobilization (e.g., mesoporous silica, sol-gel hydrogels, and porous polymer) can effectively improve enzyme stability and reduce product inhibition. However, they still face drawbacks such as potential leaching or conformational change during immobilization and poor machining performance. Especially, most enzyme carrier solid materials possess disordered structures, inevitably introducing deficiencies such as low loading capacity, hindered mass transfer, and unclear structure-property relationships. Additionally, it remains a notable challenge to meticulously design immobilization systems tailored to the specific characteristics of enzyme/reaction. Therefore, there is a significant demand for reliable solid materials to overcome the above challenges. Crystalline porous materials, particularly covalent organic frameworks (COFs), have garnered significant interest as a promising platform for immobilizing enzymes due to their unique properties, such as their crystalline nature, high porosity, accessible active sites, versatile synthetic conditions, and tunable structure. COFs create a stabilizing microenvironment that protects enzymes from denaturation and significantly enhances reusability. Nevertheless, some challenges still remain, including difficulties in loading large enzymes, reduced enzyme activities, and the limited functionality of carriers. Therefore, it is essential to develop innovative carriers and novel strategies to broaden the methods of immobilizing enzymes, enabling their application across a more diverse array of fields.The integration of enzymes with advanced porous materials for intensified performance and diverse applications is still in its infancy, and our group has done a series of pioneering works. This Account presents a comprehensive overview of recent research progress made by our group, including (i) the development of innovative enzyme immobilization strategies utilizing COFs to make the assembly and integration of enzymes and carriers more effective; (ii) rational design and construction of functional carriers for enzyme immobilization using COFs; and (iii) extensions of immobilized enzyme applications based on COFs from industrial catalysis to biomedicine and chiral separation. The integration of enzymes with functional crystalline materials offers mutual benefits and results in a performance that surpasses what either component can achieve individually. Additionally, immobilized enzymes exhibit enhanced functionality and intriguing characteristics that differ from those of free enzymes. Consistent with our research philosophy centered on integration, platform development, and engineering application, this Account addresses the critical challenges associated with enzyme immobilization using COFs while extending the applications of COFs and proposing future design principles for biomanufacturing and enzyme industry.
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Estruturas Metalorgânicas , Enzimas Imobilizadas , Polímeros , Catálise , EngenhariaRESUMO
Taxane-based chemotherapy-loaded drug delivery systems have great potential for cancer treatment. The docetaxel (DTX)-loaded PAMAM-based poly (γ-benzyl-l-glutamate)-b-d-α-tocopheryl polyethylene glycol 1000 succinate (PAM-PBLG-b-TPGS) nanoparticles and the docetaxel (DTX)-loaded PAMAM-based poly (γ-benzyl-l-glutamate) (PAM-PBLG) nanoparticles were designed using a modified nanoprecipitation method. The particle size, encapsulation efficiency (EE), and in vitro release characteristics of the nanoparticles were tested. The effects of the two nanoparticles on the cellular uptake and cell viability on human cervical cancer cell line Hela and the human breast cancer cell line MCF-7 were compared. Furthermore, their antitumor efficiency was evaluated through in vivo tumour growth experiment in comparison with free DTX. PAM-PBLG-b-TPGS nanoparticles displayed high EE, smaller diameter, and a nice releasing profile. Besides, based on the high EE and 'self-controlled' drug release of the DTX-loaded PAM-PBLG-b-TPGS nanoparticles, they exhibited stronger cytotoxicity (lower survival rate) and higher uptake rate than DTX-loaded PAM-PBLG nanoparticles in Hela cells and MCF-7 cells. Furthermore, compared with DTX-loaded PAM-PBLG nanoparticles and free DTX, DTX-loaded PAM-PBLG-b-TPGS nanoparticles produced a potent anti-tumour effect. Thus, the DTX-loaded PAM-PBLG-b-TPGS nanoparticles provide a novel attractive nanocarrier for the DTX delivery of chemotherapy to human breast cancer cells and human cervical cancer cells.
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Antineoplásicos/administração & dosagem , Neoplasias da Mama/tratamento farmacológico , Docetaxel/administração & dosagem , Portadores de Fármacos/química , Neoplasias do Colo do Útero/tratamento farmacológico , Vitamina E/química , Animais , Antineoplásicos/farmacologia , Antineoplásicos/uso terapêutico , Neoplasias da Mama/patologia , Dendrímeros/química , Docetaxel/farmacologia , Docetaxel/uso terapêutico , Liberação Controlada de Fármacos , Feminino , Células HeLa , Humanos , Células MCF-7 , Camundongos Endogâmicos BALB C , Camundongos Nus , Neoplasias do Colo do Útero/patologiaRESUMO
Diabetic kidney disease (DKD) is the most common complication of diabetes mellitus (DM). Qianjin Wenwu decoction (QWD), a well-known traditional Korean medicine, has been used for the treatment of DKD, with satisfactory therapeutic effects. This study was designed to investigate the active components and mechanisms of action of QWD in the treatment of DKD. The results demonstrated that a total of 13 active components in five types were found in QWD, including flavonoids, flavonoid glycosides, phenylpropionic acids, saponins, coumarins, and lignins. Two key proteins, TGF-ß1 and TIMP-1, were identified as the target proteins through molecular docking. Furthermore, QWD significantly suppressed Scr and BUN levels which increased after unilateral ureteral obstruction (UUO). Hematoxylin & eosin (H&E) and Masson staining results demonstrated that QWD significantly alleviated renal interstitial fibrosis in UUO mice. We also found that QWD promoted ECM degradation by regulating MMP-9/TIMP-1 homeostasis to improve renal tubulointerstitial fibrosis and interfere with the expression and activity of TGF- ß1 in DKD treatment. These findings explain the underlying mechanism of QWD for the treatment of DKD, and also provide methodological reference for investigating the mechanism of traditional medicine in the treatment of DKD.
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Nefropatias , Obstrução Ureteral , Ratos , Camundongos , Animais , Obstrução Ureteral/complicações , Obstrução Ureteral/tratamento farmacológico , Obstrução Ureteral/metabolismo , Rim/metabolismo , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Simulação de Acoplamento Molecular , Ratos Sprague-Dawley , Nefropatias/tratamento farmacológico , Matriz Extracelular/metabolismo , Flavonoides/farmacologia , Flavonoides/metabolismo , FibroseRESUMO
This paper describes the development of a film comprising chitosan (CS), sodium alginate (SA), and ethyl cellulose (EC) for buccal mucosal administration. A film of CS-SA unidirectional release drug-containing water-repellent layer EC was produced by interfacial reaction solvent-drying technique using self-made equipment. The CS-SA-EC film had superior mechanical properties compared to CS-EC and SA-EC films. The existence of the amide bond was confirmed by FT-IR. DSC confirmed that the drug was dispersed in the carrier material in an amorphous form. The drug release studies emerged that the model drugs from CS-SA-EC films presented better release properties. The Ritger-Peppas model best describes all ratios of drugs release mechanisms. The permeability characteristics of the films were evaluated in the TR146 cells model and the rabbit buccal mucosae. The cumulative penetration amounts of the model drugs were significantly increased. The permeability mechanism of the film was studied preliminarily using immunofluorescence and Western Blot. The results showed that the film inhibited the expression of ZO-1 protein, and the expressive trend of ZO-1 protein was consistent with the results of in vitro permeation experiments. The pharmacokinetics of the drugs loaded films were evaluated and compared with oral administration in rats. The relative bioavailability of the model drugs was 246.00% (Zolmitriptan) and 142.12% (Etodolac) relative to oral administration. The results of this study demonstrate the potential of CS-SA-EC vehicle in buccal mucosa drug delivery.
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Quitosana , Administração Bucal , Alginatos , Animais , Celulose/análogos & derivados , Sistemas de Liberação de Medicamentos , Mucosa Bucal , Polieletrólitos , Coelhos , Ratos , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
The purpose of the project is to establish a standardized operation method of the in vitro permeability model to maximize mucosal integrity and viability. The model drug lidocaine permeability, 20 kDa fluorescein isothiocyanate-dextran, H&E staining, and mucosal viability were used as evaluation indicators. Firstly, the buccal mucosae of rats, rabbits, dogs, porcine, and humans were analyzed by H&E staining and morphometric analysis to compare the differences. Then, we studied a series of operation methods of isolated mucosa. The buccal mucosae were found to retain their integrity in Kreb's bicarbonate ringer solution at 4 °C for 36 h. Under the long-term storage method with program cooling, freezing at -80 °C, thawing at 37 °C, and using cryoprotectants of 20% glycerol and 20% trehalose, mucosal integrity and biological viability can be maintained for 21 days. The heat separation method was used to prepare a permeability model with a mucosal thickness of 500 µm, which was considered to be the optimal operation. In summary, this study provided an experimental basis for the selection and operation of in vitro penetration models, standardized the research process of isolated mucosa, and improved the accuracy of permeability studies.
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Dextranos , Mucosa Bucal , Animais , Bicarbonatos , Cães , Glicerol , Humanos , Lidocaína , Permeabilidade , Coelhos , Ratos , Reprodutibilidade dos Testes , Solução de Ringer , Suínos , TrealoseRESUMO
The circadian clock plays a key role in our daily physiology and metabolism. Alcohol consumption disrupts the circadian rhythm of metabolic genes in the liver; however, the potential contribution of circadian clock modulation to alcoholic liver disease (ALD) is unknown. We identified a novel liver protective agent, physcion, which can alleviate fat accumulation and inflammation in ALD mice via reprogramming the hepatic circadian clock. The model of alcoholic hepatitis was established by intragastrically administering ethanol. In vitro, physcion was investigated by treating HepG2 cells with ethanol. The role of circadian clock in Physcion caused liver protection was tested by knocking down the core circadian gene Bmal1. Physcion application caused reduced lipogenesis and alleviated inflammation in alcohol-induced mice. In alcoholic hepatosteatosis models, physcion upregulated the core circadian genes. And the circadian misalignment triggered by ethanol was efficiently reversed by physcion. Physcion attenuated lipogenesis via reprogramming the circadian clock in HepG2 cells. Suppression of Bmal1 by RNA interference abolished the protective of physcion. In addition, Physcion binds to the active pocket of BMAL1 and promotes its expression. The study identified the novel liver protective effects of physcion on alcohol-induced liver injury, and modulation of the core circadian clock regulators contributes to ALD alleviation. More importantly, strategies targeting the circadian machinery, for example, Bmal1, may prove to be beneficial treatment options for this condition.
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Sirtuin 1 (SIRT1) is a protein deacetylase with important cellular functions, as it regulates numerous processes, including the circadian rhythm in peripheral tissues. Efforts are ongoing to reveal how Sirt1 can be used to treat diseases, such as alcoholic liver disease (ALD), Alzheimer's disease, and liver fibrosis. We have recently shown that noninvasive exposure to 40-Hz light flicker activates hypothalamic SIRT1 gene expression, thereby regulating the central circadian clock. This study investigated the effects of 40-Hz light flicker in a mouse model of ALD. RNA sequencing (RNA-seq) analysis was performed to explore the potential pathways affected by 40-Hz light flicker. We found that 40-Hz light flicker significantly decreased the acute ethanol-induced increases in serum alanine aminotransferase (ALT) and serum triglyceride (TG) levels and reduced fat-droplet accumulation in mouse livers. Additionally, 40-Hz light flicker significantly suppressed ethanol-induced increases in sterol regulatory element binding protein 1 (SREBP-1) and fatty acid synthase (Fasn) levels. Furthermore, the ethanol induced significant decreases in both Sirt1 levels and phosphorylation of adenosine monophosphate-activated protein kinase subunit (AMPKα), compared with those in the control group. Strikingly, pretreatment with 40-Hz light flicker ameliorated such ethanol-induced decreases in SIRT1 levels and AMPKα phosphorylation. In addition, ethanol-induced increases in levels of brain and muscle arnt-like protein-1 (BMAL1), circadian locomotor output cycles kaput (CLOCK), and period 2 (PER2) were reversed by 40-Hz light flicker. RNA-seq analysis revealed significant differences in expression of genes related to the AMPK signalling. Moreover, ethanol consumption altered mRNA levels of Sirt1 and circadian genes in the suprachiasmatic nucleus (SCN), indicating that ethanol influenced central pacemaker genes; however, 40-Hz light flicker reversed these ethanol-induced changes. Taken together, our findings demonstrate that 40-Hz light flicker rapidly influence the SCN and exhibits inhibitory properties on hepatic lipogenesis, indicating that 40-Hz light flicker has therapeutic potential for preventing alcoholic liver steatosis.
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Liver inflammation and excessive accumulation of lipids play a critical role in alcoholic liver diseases (ALD) pathogenesis. Plant polyphenols are widely used to prevent toxic liver damage. The anthocyanin from Lonicera caerulea L. was extracted and purified. The aim of the study was to evaluate the hepatoprotective mechanism of the purified component (PLE), focusing on the effects of PLE on alcoholic steatohepatitis. C57BL/6 mice were fed on chronic plus binge ethanol in Lieber-DeCarli liquid diet to establish acute ethanol model. PLE treatment significantly reduced the accumulation of serum aminotransferase and triglycerides and increased albumin levels in ethanol-induced mice. Also, PLE ameliorated histological changes and lipid droplets induced by ethanol. In addition, PLE obviously suppressed the expression of SREBP1 and enhanced phosphorylation of AMPK compared with chronic ethanol administration. PLE suppressed inflammasome activation by decreasing F4/80 level and inhibiting caspase-1, thereby preventing activated macrophages from producing pro-inflammation cytokines. AML12 cells were pretreated with different concentrations of PLE for 2â¯h and then exposed to ethanol for 48â¯h. PLE suppressed the expression of SREBP1 and enhanced phosphorylation of AMPK in AML12 cells exposed to ethanol. Additionally, PLE inhibited the expression of F4/80 and decreased IL-1ß release. AMPK interference confirms that PLE downregulation SREBP1 and F4/80 depending on AMPK activation in ethanol-treated AML12 cells. PLE possessed the capacity for inhibiting the inflammatory response and suppressing lipid accumulation, indicating that PLE can be used as a dietary health supplement for alcoholic steatohepatitis.