APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2022
·Vol. 106
·期 23
·7805-7817
·
IF 5.56
Fu Linglong,Sun Mengning,Wen Weizhang,Dong Na,Li Defa
Abstract Small ubiquitin-like modifier (SUMO) tag is widely used to promote soluble expression of exogenous proteins, which can then be cleaved by ubiquitin-like protease 1 (Ulp1) to obtain interested protein. But the application of Ulp1 in large-scale recombinant protein production is limited by complicated purification procedures and high cost. In this study, we describe an efficient and simple method of extracellular production of Ulp1 403-621 using a leaky Escherichia coli BL21(DE3), engineered by deleting the peptidoglycan-associated outer membrane lipoprotein ( pal ) gene. Ulp1 403-621 was successfully leaked into extracellular supernatant by the BL21(DE3)-Δ pal strain after IPTG induction. The addition of 1% glycine increased the extracellular production of Ulp1 403-621 approximately four fold. Moreover, extracellular Ulp1 403-621 without purification had high activities for cleaving SUMO fusion proteins, and antimicrobial peptide pBD2 obtained after cleavage can inhibit the growth of Staphylococcus aureus . The specific activity of extracellular Ulp1 403-621 containing 1 mM EDTA and 8 mM DTT reached 2.0 × 10 6 U/L. Another commonly used protease, human rhinovirus 3C protease, was also successfully secreted by leaky E. coli strains. In conclusion, extracellular production of tool enzymes is an attractive way for producing large-scale active recombinant proteins at a lower cost for pharmaceutical, industrial, and biotechnological applications. Key points
• First report of extracellular production of Ulp1
403-621
in leaky Escherichia coli BL21(DE3) strain.
• One percent glycine addition into cultivation medium increased the extracellular production of Ulp1
403-621
approximately four fold.
• The specific activity of extracellular Ulp1
403-621
produced in this study reached 2.0 × 10
6
U/L.
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CHEMICO-BIOLOGICAL INTERACTIONS
·
2014
·
IF 2.982
Lihua Hu,Lingrui Li,Demei Xu,Xiaomin Xia,Ruxian Pi,Duo Xu,Wenchao Wang,Hong Du,Erqun Song,Yang Song
The purpose of this study was to investigate the possible hepatoprotective effects of neohesperidin dihydrochalcone (NHDC) on carbon tetrachloride (CCl 4 )-induced acute oxidative injury in vivo and in vitro . In a mouse model, intraperitoneal injection of CCl 4 resulted in a significant increase in serum aspartate transaminase (AST) and alanine transaminase (ALT) activities. Histopathological examination revealed severe hepatocyte necrosis and destruction of architecture in liver lesions, and immunohistochemical staining illustrated a remarkable enhancement of COX-2 and iNOS expression. The levels of hepatic antioxidant, such as, catalase (CAT), total superoxide dismutase (T-SOD), glutathione peroxidase (GP-X) and glutathione (GSH) were decreased, compared to the control group. However, pretreatment of NHDC for six consecutive days significantly ameliorated these changes. Moreover, Western blotting assay indicated pretreatment with NHDC also down-regulated CCl 4 -induced protein expressions of NF-κB, IL-6, caspase 3 and caspase 8. In HepG2 cell model, CCl 4 -treatment caused significant decrease in cell viability, antioxidant activities and GSH level, increase in intracellular reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS) level. Interestingly, pretreatment of NHDC effectively relieved CCl 4 -induced oxidative damage in a dose-dependent manner. In conclusion, NHDC appeared to possess promising anti-oxidative and anti-inflammatory capacities, it is possible to be used as a hepatoprotective agent.
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ALLERGY
·
2021
·Vol. 77
·期 2
·483-498
·
IF 13.146
Renlan Wu,Xiefang Yuan,Xingjie Li,Ning Ma,Hongyu Jiang,Hongmei Tang,Guofeng Xu,Zhigang Liu,Zongde Zhang
Background Alteration of commensal microbiota is highly correlated with the prevalence of allergic reactions to food in the gastrointestinal tract. The mechanisms by which microbiota modulate food allergen sensitization in the mucosal site are not fully understood. Methods We generate DCs specific knockout of retinoic acid receptor α (Rara) gene mice (DC KO Rara) to evaluate food sensitization. The bile acid-activated retinoic acid response was evaluated by flow cytometry, real-time RT-PCR and Illumina transcriptome sequencing. The global effect of Abx treatment on BA profiles in the mucosal lymph tissue mLN in mice was examined by UPLC-MS analysis. Results In this study, we demonstrate that depletion of commensal gut bacteria leads to enhanced retinoic acid (RA) signaling in mucosal dendritic cells (DCs). RA signaling in DCs is required for the production of food allergen-specific IgE and IgG1. Antibiotics induced an enlarged bile acid (BA) pool, and dysregulated BA profiles contributed to enhanced RA signaling in mucosal DCs. BA-activated RA signaling promoted DC upregulation of interferon I signature, RA signature, OX40L, and PDL2, which may lead to T helper 2 differentiation of CD4+ T cells. BA-activated RA signaling involved the farnesoid X receptor and RA receptor α (RARa) interaction. Depletion of bile acid reduces food allergen specific IgE and IgG1 levels in mice. Conclusion Our research unveils a mechanism of food sensitization modulated by BA-RA signaling in DCs, which suggests a potential new approach for the intervention of food allergic reactions.
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INTERNATIONAL JOURNAL OF PHARMACEUTICS
·
2020
·
IF 4.845
Liqiong Zha,Jiajia Qian,Beilei Wang,Huanhuan Liu,Caiyun Zhang,Qiannian Dong,Weidong Chen,Lufeng Hong
As one of the active pharmaceutical ingredients in Gamboge, Gambogenic acid (GNA) has shown diverse anti-tumor activities. To reduce the vascular irritation of GNA and improve its water solubility, tumor targeting, and bioavailability, GNA loaded Zein nanoparticles ( [email protected] NPs) was further coated by polydopamine (PDA) to develop [email protected] NPs by anti-solvent precipitation and surface modification. The results showed that particle size and Zeta potential of [email protected] NPs were about 310 nm and −40.8 mV with core–shell morphology confirmed by TEM. [email protected] NPs increased the water solubility of GNA by more than 700 times and showed pH-sensitive release behavior in PBS with pH 6.86. In vitro cytotoxicity tests showed that [email protected] NPs had higher inhibitory activity on HepG2 cells than free GNA, and their IC 50 were 1.59 μg/mL and 9.89 μg/mL, respectively. Additionally, the hemolysis and vascular irritation assay showed that [email protected] NPs had good cytocompatibility and reduced the irritation of GNA to blood vessels. Moreover, the in vivo pharmacokinetic experiments exhibited that the C max and AUC 0-t of [email protected] NPs were significantly improved approximately by 2.09-fold and 3.48-fold over that of GNA, respectively. In conclusion, [email protected] NPs solve many defects of GNA and provide a tumor-targeting drug delivery for GNA.
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Drug Design Development and Therapy
·
2022
·
IF 4.319
Yanping Wu,Gen-You Liao,Hua-Jing Ke,Pi Liu
Objective We aimed to investigate the effects of snake-derived phospholipase A2 inhibitor (PLA2) from Sinonatrix percarinata and Bungarus multicinctus on acute pancreatitis in vivo and in vitro and assess the mechanisms.Methods The levels of platelet-activating factor (PAF) and tumor necrosis factor (TNF)-α were detected by ELISA, and the characteristics of autophagy were detected by transmission electron microscopy and Western blotting (LC3, p62, and ATG5).Results In vitro experiments showed that PLA2 treatment caused obvious formation of autophagic bodies. By contrast, Sinonatrix and Bungarus peptides reduced the number of autophagic bodies. The concentrations of PAF and TNF-α, and the expressions of p62, autophagy-related 5 (ATG5), and microtubule-associated protein 1A/1B-light chain 3 (LC3)II/LC3I in the PLA2-treated group were significantly higher than in the control group (P<0.05). The concentrations of PAF and TNF-α, and the expressions of p62, ATG5, and LC3II/LC3I in the Sinonatrix or Bungarus peptide treatment groups were significantly lower than in the PLA2-treated cells (P<0.05). In the pancreatic tissue, autophagic bodies were observed in the model group; autophagic bodies were remarkably reduced in Sinonatrix or Bungarus peptide-treated groups compared with the model group. In vivo experiments also showed that the levels of PAF and TNF-α, and the expressions of p62, ATG5, and LC3II/LC3I were significantly higher in the model group than in the control group (P<0.05). The levels of PAF and TNF-α in the model group, and the expressions of p62, ATG5, and LC3II/LC3I in Sinonatrix or Bungarus peptide-treated groups were significantly lower than in the model group (P<0.05).Conclusion Sinonatrix or Bungarus peptide could ameliorate the features of acute pancreatitis, likely through regulating autophagy.
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Journal of Building Engineering
·
2025
·
IF 7.4
Lei Fang,Zhixiang Liu,Jiaqi Zhou,Ju Lin,Song Xu,Xiaojuan Jia
To mitigate the inherent brittleness of cement-based materials, this research introduces a novel in-situ polymerization method using acrylamide (AM) monomer and ammonium persulfate (APS) as the initiator. Mechanical testing and multi-scale analysis (SEM/FTIR/XRD) indicate that incorporating 3 % AM content improves the toughness of cement pastes by 47 % while retaining about 94 % of their initial compressive strength. This improvement arises from the formation of organic-inorganic interpenetrating networks. Early-stage performance is crucially regulated by optimized APS initiator dosage: 1 % APS guarantees full AM conversion (≤3 % AM), whereas 5 % APS accelerates PAM network formation and increases 3-day toughness by 22 % through accelerated skeletal framework development. The water released from the PAM hydrogel initially formed by AM in-situ polymerization facilitates subsequent cement hydration and promotes the reconstruction of the polymer network, which together contributes to the substantial toughening observed at later stages. The results of this study provide important insights for designing in-situ polymerized cement-based materials with greater toughness.
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Pain Physician
·
2022
·Vol. 25
·期 8
·E1239
·
IF 4.396
Li-Hong Sun,Qi Xu,Lin Jin,Li-Dan Jin,Qing Chen,Hui Wu,Xin-Zhong Chen
Background: Morphine is one of the preferred drugs for the clinical treatment of pain. Both clinical and preclinical studies have reported sexual dimorphism in morphine analgesia. Different circulating levels of estrogen could be involved in sex differences in response to morphine analgesia. In our previous research, we found that capsaicin injection into the cervix of rats caused acute visceral pain that could be relieved by morphine. The role of estrogen in morphine analgesia in rats under uterine cervix pain and its underlying mechanisms remain to be explored. Objectives: The present study aims to investigate the effect of estrogen on morphine analgesia and its underlying mechanism in rats under uterine cervix pain. Study design: Controlled animal study. Setting: University laboratory. Methods: First, we compared the analgesic effect of morphine in ovariectomized rats with uterine cervix pain with or without estrogen replacement. Then, the changes in the expression of opioid receptors and L-type voltage-gated calcium channels (L-type-VGCC, LTCC) at the spinal level were detected by real-time quantitative polymerase chain reaction. Finally, we investigated the effect of the manipulation of spinal LTCC (L-type CaV1.2 calcium channel, L-type CaV1.3 calcium channel) on the estrogen-mediated inhibition of morphine analgesia. Results: Our study shows that morphine antinociception is diminished in rats with uterine cervix pain that are treated with estrogen. Estrogen treatment increases the expression of spinal CaV1.2 and CaV1.3, while only anti-CaV1.2 treatment impaired estrogenic suppression of morphine antinociception. Limitations: More underlying mechanisms of the role of spinal CaV1.2 in modulating estrogen-mediated inhibition of morphine analgesia need to be explored in future research. Conclusions: This is the first evidence that spinal CaV1.2 is involved in estrogenic modulation of morphine antinociception in rats under uterine cervix pain. Our results will provide new ideas and references for estrogen-related differential prescription of opioids.
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Biomedical Materials
·
2022
·Vol. 17
·期 6
·065020
·
IF 4.103
Yanyun Pang,Dan Li,Jing Zhou,Xue Liu,Min Li,Yanling Zhang,Daixing Zhang,Xu Zhang,Qing Cai
Native bone tissue can be formed by developing collagen fibrils coated with hydroxyapatite (HA) and whitlockite (WH) nanoparticles after mineralization. WH has attracted much attention as the second most abundant bone mineral in human bones. It has a negatively charged surface, which can adsorb osteogenesis-related proteins such as bone sialoprotein in vivo, thus having a stronger possibility to induce osteogenesis. However, due to its poor thermodynamic stability and intermediate phases, the preparation of WH is relatively tricky, so WH inorganic scaffolds are still rarely studied. Therefore, this study explored the preparation of WH inorganic scaffolds using the hydrothermal method and prepared pure inorganic WH scaffolds. The prepared scaffolds exhibited apparent WH crystal phases in the x-ray powder diffraction (XRD) characterization. In the scanning electron microscopy (SEM) images, the WH scaffolds had an apparent hexagonal crystal form, which had a pronounced effect on promoting cell proliferation and differentiation in vitro experiments compared to the HA and HA/WH scaffolds. Furthermore, the scaffolds were used to verify the osteogenic properties of subcutaneous ectopic osteogenesis or repair of the calvarial defect in vivo and proved that the WH inorganic scaffolds have an excellent synergistic osteogenic ability.
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Biomed Research International
·
2020
·
IF 2.276
Lei Xiao,Xia Zhang,Zhiyi Chen,Jianhua Li,Bing Li,Lin Li
Hair loss is a common disease in dermatology, while the approved drugs may have unpredictable side effects. In this study, the effect of timosaponin BII extracted from Anemarrhena asphodeloides on hair growth of C57BL/6 mice was investigated by measuring the hair follicle morphology, hair growth length and area in C57BL/6 male mice, and the immunohistochemical analysis of β-catenin, Wnt3a, and Wnt10b in the dorsal skins of mice after topical application with minoxidil and timosaponin BII for 15 days. The decrease in skin brightness, the increase in the regrowing area of hair and hair follicles numbers, and the improvement of hair follicle morphology in the group applied with 0.5% timosaponin BII indicated an induction of the anagen phase in telogenic mice skin, which were comparative to the 2% minoxidil treatment. The immunohistochemical analysis detected an increase in the expression of β-catenin and Wnt10b, supporting the theory of the activation of the β-catenin/Wnt pathway was one of the pathways that are related to anagen phase induction. Anemarrhena asphodeloides is a herb commonly used for metabolic disorders in China. The present study is the first to show that the timosaponin BII, which is present at a high concentration in A. asphodeloides, promotes hair growth in C57BL/6 male mice. The results indicate that timosaponin BII may be a potential promoting agent for hair growth.
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Food & Function
·
2020
·Vol. 12
·期 2
·791-801
·
IF 4.171
Youyou Lu,Zhuang Yu,Zhe Zhang,Xi Liang,Pimin Gong,Huaxi Yi,Liuqing Yang,Tongjie Liu,Hanping Shi,Lanwei Zhang
Probiotics and natural products can promote humoral transport and effectively relieve intestinal motility. This study investigated the effects of probiotics in combination with konjac glucomannan (KGM) and an aqueous extract of Prunus persica on constipation. The growth promotion effect of these natural products on probiotics was investigated using co-culture in vitro. The combined effect of probiotics and natural products on constipation was observed in mice. The tryptophan, tryptamine and short-chain fatty acid levels were determined using enzyme-linked immunosorbent assay, reverse-phase high-performance liquid chromatography, and gas chromatography. The key genes and proteins involved in humoral transport were identified using real-time polymerase chain reaction, western blotting and fluorescence immunoassay. KGM promoted the growth of Bifidobacterium animalis F1-7 in vitro, and a mixture of KGM and B. animalis F1-7 effectively promoted defaecation in mice, increased the faecal water content, shortened the defaecation time and improved the gastrointestinal transit rate. In mice, the KGM + F1-7 mixture reduced the tryptophan level and increased the levels of tryptamine, acetic acid, propionic acid, butyric acid and valeric acid. In addition, the KGM + F1-7 mixture effectively increased the mRNA level of 5-HT4-G-protein-coupled receptors (5-HT4GPCR)/mucins-2 (MUC-2) and reduced the level of aquaporins (AQP3); furthermore, it upregulated the protein level of 5-HT4GPCR/MUC-2 and downregulated the protein level of AQP3. These findings indicated that the KGM + F1-7 mixture effectively improved intestinal motility and alleviated constipation through humoral transport-related pathways.
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Gastroenterology Research and Practice
·
2020
·
IF 1.806
Geng Chen,Shuodong Wu
This study was aimed at investigating the effect of baicalin on experimental cholesterol gallstones in mice. The mouse gallstone model was induced by feeding with a lithogenic diet, and cholesterol stones were found in the gallbladder. The lithogenic diet caused elevation of triglycerides, cholesterol, and low-density lipoprotein concentrations and descent of high-density lipoprotein concentration in serum. Hyperplasia and inflammatory infiltration were observed in the gallbladder wall of lithogenic diet-fed mice. We also found the increase of cholesterol content and the decrease of bile acid in bile. Real-time PCR and western blot results demonstrated that the expression levels of two enzymes (cholesterol 7α-hydroxylase (CYP7a1) and sterol 12α-hydroxylase (CYP8b1)) to catalyze the synthesis of bile acid from cholesterol were decreased and that two cholesterol transporters (ATP-binding cassette transporter G5/G8 (ABCG5/8)) were increased in the liver of lithogenic diet-fed mice. The lithogenic diet also led to enhanced activity of alanine aminotransferase and aspartate aminotransferase in serum; increased concentrations of tumor necrosis factor-α, interleukin- (IL-) 1β, IL-6, and malondialdehyde; and decreased superoxide dismutase activity in the liver, suggesting inflammatory and oxidative stress. In addition, liver X receptor alpha (LXRα) was increased in the liver. After gavage of baicalin, the lithogenic diet-induced gallstones, hyperlipidemia, gallbladder hyperplasia, inflammation, and oxidative stress in liver and cholesterol metabolism disorders were all alleviated to some degree. The expression of LXRα in the liver was inhibited by baicalin. In addition, the LXRα agonist T0901317 aggravated lithogenic diet-induced harmful symptoms in mice, including the increase of gallstone formation, hyperlipidemia, hepatic injury, inflammation, and oxidative stress. In conclusion, we demonstrated that baicalin played a protective role in a lithogenic diet-induced gallstone mouse model, which may be mediated by inhibition of LXRα activity. These findings may provide novel insights for prevention and therapy of gallstones in the clinic.
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Evidence-based Complementary and Alternative Medicine
·
2021
·
IF 2.63
Meiyu Zhang,Yizhu Wang,Yongxi Wu,Fangtong Li,Mingxin Han,Yulin Dai,Fei Zheng,Hao Yue
Protopanaxadiol (PPD)-type ginsenosides are the main ginsenosides in ginseng (Panax ginseng C. A. Meyer) with potential therapeutic effects on diseases related to intestinal flora imbalance. This study aimed to investigate the in vitro metabolism of protopanaxadiol ginsenosides in human intestinal flora and their effect on the flora. Rapid resolution liquid chromatography coupled with quadruple-time-of-flight mass spectrometry (RRLC-Q-TOF MS) was utilised for the transformation of ginsenoside constituents for sample identification. Using 16S rDNA gene sequencing technique, the effect of PPD-type ginsenosides on gut microflora was analysed based on the indices of microflora diversity and gut microflora. The sample was transformed for 6 h, and the metabolites were ginsenoside Rb1, Rc, Rb2, Rb3, CO, Gyp-IX, Gyp-XVII, CMc-1, F2, Rg3, CK, Rh2, and PPD. The metabolites were CK, Rh2, and PPD when the samples were transformed for 60 h. The intestinal microflora were subjected to high-throughput sequencing using the Illumina MiSeq 2500 sequencing platform. In comparison with the faecal sample from the blank group, the protopanaxadiol saponin group significantly increased the relative abundance of Firmicutes and significantly decreased Bacteroidetes and Proteobacteria at the phylum level, whereas it significantly increased the relative abundance of Prevotella_9, Faecalibacterium, and Dialister and significantly decreased Escherichia-Shigella, Dorea, and Lachnoclostridium at the genus level. This study provides a basis for the determination of the pharmacodynamic material basis and pharmacodynamic targets of PPD-type ginsenosides based on the intestinal flora.
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